Prospecting and characterization of potential probiotic Lactobacillus species from animal gut and food sources

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This study aims to prospect and characterize Lactobacillus species with probiotic potential from animal gut, fruit, and vegetable sources. Method: Lactobacillus species were isolated from samples using selective enrichment techniques. Colony morphology, catalase testing, and Gram staining were used for the preliminary identification of the presumptive Lactobacillus strains. Their identities were subsequently confirmed by 16S rRNA gene sequencing and BLAST analysis. The identified Lactobacillus strains were subjected to in vitro characterization to assess their probiotic properties. Results: Ten Lactobacillus species isolates demonstrated high acid tolerance, with survivability exceeding 50% at pH 2.5 and 3.0. Their survival rates ranged from 62.05% to 97.42% and from 81.12% to 99.31% in the presence of 0.5% and 0.3% bile salt concentrations, respectively. All the isolates exhibited varying capacities to reduce cholesterol (13.06% –57.64%) and demonstrated antioxidant activity ranging from 6.81% to 24.45%. Only one isolate (Mcxc) lacked bile salt hydrolase activity, and 33.33% of the isolates did not produce any exopolysaccharides. The auto-aggregation abilities ranged from 18% to 52% after 4 h of incubation, while cell surface hydrophobicity ranged from 0.00% to 84.68%. None of the isolates exhibited haemolytic or DNase activity. The degree of adhesion to Caco-2 cells ranged from 2.6% to 15.29%. All isolates were susceptible to erythromycin, chloramphenicol, ampicillin, and penicillin G. Notably, only four Lactobacillus isolates exhibited inhibitory activity against all the tested pathogenic bacteria. Conclusions: Our findings revealed that Lactiplantibacillus plantarum To3a and To3d, isolated from tomatoes, stand out as a viable and alternative source of probiotics due to their promising probiotic properties and health-promoting benefits; thus, they may be explored for practical use in novel, locally adaptable functional products. Lactobacillus probiotic fruits vegetables animal gut food sources Figures Figure 1 Figure 2 Background Lactobacillus is a prominent member of lactic acid bacteria (LAB), which are Gram-positive, rod-shaped, microaerophilic bacilli , with over 290 species having catalase-and oxidase-negative features ( 1 ). They include, among others, Limosilactobacillus reuteri , Lactobacillus acidophilus , Lacticaseibacillus casei , Lacticaseibacillus rhamnosus , Limosilactobacillus fermentum , Levilactobacillus brevis , Lactiplantibacillus plantarum , Ligilactobacillus salivarius , and Lacticaseibacillus paracasei ( 2 , 3 ). Lactobacillus species are typically found in diverse niches, including the gastrointestinal tract (GIT) of humans and animals, plant surfaces, fermented foods, water, wine, soil, and sewage, among others ( 4 , 5 ). These bacteria play a considerable role in food fermentation, flavour, texture, and aroma development in food products ( 6 ), and they are "generally recognized as safe"(GRAS) and have a qualified safety estimate (QPS) status ( 7 ). Hence, it may be safely exploited for extensive probiotic application. Some Lactobacillus species are probiotics, live microorganisms, which, when consumed in sufficient amounts, benefit the host's health ( 8 ). The health benefits of probiotics, which include intestinal microbiota balance, intestinal tract relief, allergy alleviation, obesity, diabetes, bacterial vaginosis, and oral diseases, have been reported in many studies ( 3 , 9 , 10 ). In addition to those mentioned above, other health advantages, such as the prevention and management of cancer, hypercholesterolemia, lactose intolerance, improved immune response, inflammatory bowel disease, and irritable bowel syndrome, have been reported ( 11 , 12 ). Consequently, these valuable microbes have gained extensive recognition and research interest over the years due to their broad spectrum of positive health effects ( 13 ). Many probiotic strains have been successfully isolated from fruits and vegetables ( 14 ), fruits and fermented foods ( 15 ), leaves of food plants ( 16 ), chickens ( 17 – 20 ), the GIT of snakehead fish ( 21 ), the GIT of wild boar ( 22 ), and the intestinal mucosa of healthy piglets ( 23 ). For example, Lactobacillus acidophilus CM1 and Lactobacillus delbrueckii OS1 strains isolated from different food sources ( 24 ) and Lactiplantibacillus plantarum strains isolated from silages from different forage plants, artisanal salami, and fermented cocoa beans demonstrated promising probiotic properties ( 25 ). Notably, the isolation of these strains from various food sources has great potential because of their naturalness as well as the techno-functional and health benefits they may possess ( 26 ). While probiotics offer well-documented health benefits, accessibility and affordability remain significant challenges in developing countries, especially in rural areas ( 27 , 28 ). Most commercially available probiotic products in Africa are based on Lactobacillus strains sourced from countries outside the continent, rather than utilizing indigenous species from the African microbiota ( 8 ), with bacterial strains such as Lacticaseibacillus paracasei Shirota, Lacticaseibacillus rhamnosus GG, and Limosilactobacillus reuteri RC-14 recognized as the most commercially available and established probiotics ( 16 , 29 , 30 ). Unfortunately, some of these imported probiotic cultures are expensive for many African locals ( 31 ), thus, hampering the development of innovative food products from locally sourced probiotic organisms. In response to the limited representation of locally sourced probiotic strains in commercial formulations, this study aimed to prospect, isolate, and characterize Lactobacillus species from animal GIT, as well as from fruit and vegetable sources, within the Johannesburg metropolitan area of South Africa. Materials and methods Sample collection Fruit (bananas) and vegetables (tomatoes and cucumbers) were purchased from local street stalls and retail stores in the central business district (CBD) of Johannesburg, Gauteng Province, South Africa. In contrast, fresh chicken guts were purchased from live poultry stores in the CBD of Johannesburg, Gauteng Province, South Africa. The samples were aseptically collected in sterile plastic containers. They were immediately transported in a cooler ice box to the microbiology laboratory for further analysis within 24 h. Enrichment and isolation of Lactobacillus species Fruit and vegetable samples were washed separately with sterile distilled water. Banana fruit (n = 5) or vegetable samples (n = 5) were aseptically cut on sterile aluminium foil into separate small pieces, after which, one gram (1 g) from each dissected fruit or vegetable sample was separately homogenized in 9 milliliter (ml) of 0.1% sterile peptone solution (Merck, SA) using a sterile Waring laboratory blender for 2 min. Broiler chicken caecal (n = 3) and small intestine (n = 3) portions were aseptically dissected separately to obtain the contents inside. Then, 1 g of each caecal or small intestine content was separately homogenized in 9 ml of peptone solution. Following homogenization, each sample was serially diluted up to 10⁷–fold with buffered peptone water. Subsequently, 1 ml from each dilution was aseptically spread onto De Man, Rogosa, and Sharpe (MRS) agar plates (Merck, South Africa) and incubated anaerobically at 37°C for 24 to 48 h. Thereafter, a single colony of each Lactobacillus isolate, showing different morphologies from each other, was carefully picked and streaked onto a newly prepared MRS agar plate to obtain pure bacterial cultures. These cultures were then harvested and stored at – 80°C in MRS broth containing 20% sterile glycerol (v/v) for further analyzes. Identification of lactobacilli by 16S rRNA gene sequencing Lactobacillus species isolates were subjected to Gram staining, motility, and catalase tests as described by Hidayat et al. ( 32 ). Motile, catalase-negative, Gram-positive, rod-shaped isolates were subjected to molecular characterization. Genomic DNA was extracted using the ZR Fungal/Bacterial DNA Miniprep™ Kit (Zymo Research, Irvine, USA). The DNA was quantified using a NanoDrop™ One UV-Vis Spectrophotometer (Thermo Fisher Scientific, USA). The 16S rRNA genes of the isolates were amplified using a set of universal primers 27F (5’-AGAGTTTGATCCTGGCTCTCAG-3’) and 1429R (5’-GGTTACCTTGTTACGACTT-3’). The polymerase chain reaction (PCR) was performed in a 25 µl volume, which consisted of 12.5 µl of 2x PCR master mix (Anatech, SA), 1 µl each of the forward and reverse primers, 1 µl of chromosomal DNA, and nuclease-free water to make up the final volume. PCR amplification was performed using Bio-Rad T100 TM Thermocycler under the following program: pre-denaturation for 3 min at 95°C, followed by 35 cycles of denaturation (95°C, 30 s), annealing (55°C, 55 s), extension (72°C, 60 s) and final extension at 72°C for 10 min. The quality of the PCR product was examined on a 1.5% (w/v) agarose gel using the Mini-Sub® Cell GT (Bio-Rad Laboratories Inc., California, USA) and visualized using the Gel DOC™ XR + Imaging system after staining with ethidium bromide. PCR purification and sequencing of the amplified PCR products were conducted by Inqaba Biotechnology Company, Pretoria, SA. The forward and reverse 16S rRNA gene sequences of the isolates were edited using BioEdit Sequence Alignment Editor version 7.2 software. The identity of the bacterial strains was determined using the Basic Logical Alignment Search Tool (BLAST) program ( https://blast.ncbi.nlm.nih.gov/Blast.cgi ). Screening for Probiotic properties of bacterial isolates Preparation of Lactobacillus cell suspensions Pure cultures of each Lactobacillus species from MRS agar plates were inoculated into 10 ml MRS broth and incubated at 37°C for 18 h to obtain a pure culture. One ml of each culture was centrifuged at 10000 rpm for 5 min at 4°C. The cell pellets were washed twice with phosphate-buffered saline (PBS) at pH 7.2, and the washed pellets were used to prepare cell cultures for subsequent analysis. For each experimental assay, the optical density (OD) at 600 nm of the bacterial suspensions was adjusted to either 1.0 (for 2,2-diphenyl-1-picrylhydrazyl scavenging and cell adhesion activity) or 0.25 (for antimicrobial activity, cell auto-aggregation, cell surface hydrophobicity, antibiotic susceptibility, and DNase activity) using a Genesys 10S VIS spectrophotometer (Thermo Scientific, USA). Acid tolerance test Each pure culture of Lactobacillus was adjusted to 10 9 CFU/ml and used for analyzes. The acid tolerance of the selected Lactobacillus isolates was examined by the method described by Kumar et al. ( 33 ). The acidic medium was prepared by adjusting the MRS broth with 1 N hydrochloric acid (HCl) (Sigma, SA) to a pH of 2.5 or 3.0. Approximately 0.1 ml of each Lactobacillus cell suspension at 10^9 CFU/ml was inoculated into 10 ml of acidic MRS broth at pH 2.5 and pH 3 and incubated anaerobically at 37°C for 0 and 3 h. Next, 0.1 ml of each of the serial dilutions (10 1 – 10 6 ) of the acid medium culture was spread on an MRS agar plate, followed by anaerobic incubation for 48 h at 37°C. The survival rate of Lactobacillus isolates under low acidic conditions was calculated using the formula described by Dehghani et al. ( 34 ). Lacticaseibacillus rhamnosus GG (Anatech, SA) was used as a positive control. Survival rate % = \(\:\frac{\text{l}\text{o}\text{g}\text{C}\text{F}\text{U}\:\text{o}\text{f}\:\text{v}\text{i}\text{a}\text{b}\text{l}\text{e}\:\text{c}\text{e}\text{l}\text{l}\text{s}\:\text{s}\text{u}\text{r}\text{v}\text{i}\text{v}\text{e}\text{d}\:}{\text{l}\text{o}\text{g}\text{C}\text{F}\text{U}\:\text{o}\text{f}\:\text{i}\text{n}\text{i}\text{t}\text{i}\text{a}\text{l}\:\text{c}\text{e}\text{l}\text{l}\text{s}\:\text{i}\text{n}\text{n}\text{o}\text{c}\text{u}\text{l}\text{a}\text{t}\text{e}\text{d}}\) \(\:\times\:\) 100 Bile tolerance test With minor modifications, the method described by Kumar et al. ( 33 ) was used to test the bile salt tolerance of the Lactobacillus isolates. Approximately 0.1 ml of each Lactobacillus cell suspension at 10 9 CFU/ml was inoculated into 10 ml of MRS broth containing 0.3% or 0.5% (w/v) bile salt (Sigma-Aldrich) and incubated at 37°C for 0 and 4 h under anaerobic conditions. After incubation, 0.1 ml of each serial dilution (10 1 – 10 6 ) was spread on MRS agar plates and incubated anaerobically at 37°C for 48 h. The survival rate was calculated using the formula applied for the acid tolerance. Lacticaseibacillus rhamnosus GG was used as a positive control. Bile salt hydrolase (BSH) assay The BSH activity of the isolates was tested qualitatively using the direct plate BSH method described by Habib et al. ( 35 ). MRS agar plates supplemented with calcium chloride (CaCl 2 ) (0.37 g/L) (Merck, SA) and sodium taurocholate (TDCA) (0.5%, w/v) (Merck, SA) were freshly prepared, and wells were created on the agar plates with sterile blue P1000 pipette tips (SBPT). Thereafter, 0.1 ml of each Lactobacillus culture was pipetted into the wells of the supplemented media plates and incubated anaerobically at 37°C for 48 h. Agar plates without CaCl 2 and TDCA were used as controls. The precipitation of bile acids around the wells indicated BSH activity. BSH activity was expressed based on the diameter of the precipitation zones around the wells. It was evaluated as follows: no precipitation (−), precipitation zone (+) up to 10 mm, precipitation zone 10–15 mm (++), and precipitation zone > 15 mm (+++). Cholesterol-lowering ability of Lactobacillus isolates The in vitro cholesterol -lowering ability assay was conducted using the o -phthalaldehyde method detailed by Srinivash et al. ( 36 ). A syringe filter (0. 45 µm) was used to filter water-soluble cholesterol (10 mg/100 ml; Merck, SA). Then,100 µl of this water-soluble cholesterol was mixed with 9.9 ml of MRS broth supplemented with 0.3% bile salt. The mixture was inoculated with 1 ml of a suspension (10^8 CFU/ml) of each Lactobacillus isolate and incubated at 37°C for 24 h. The MRS broth without 0.3% bile salt and water-soluble cholesterol served as the control medium. After incubation, the cells were pelleted by centrifugation at 8000 × g for 10 min. One ml of the supernatant was added to a sterile test tube, along with 1 ml of potassium hydroxide (33% w/v; Merck, SA) and 2 ml of 95% absolute ethanol. The mixture was vortexed for 1 min, incubated at 35°C for 15 min, and then placed in a water bath at 37°C for an additional 15 min. After cooling to room temperature, 2 ml of distilled water and 3 ml of hexane (Sigma, SA) were added, and the mixture was vortexed for 1 min. The mixture was then allowed to stand for 15 min to facilitate phase separation. One ml of the hexane layer was transferred into a sterile glass vial and heated in a water bath at 65°C to evaporate the hexane solvent. After hexane evaporation, 2 ml of o-phthalaldehyde reagent (Merck, SA) was added to the residue. After thorough mixing, 0.5 ml of concentrated sulfuric acid (Sigma, SA) was carefully added, and the mixture was vortexed for 1 min. The resulting mixture was allowed to stand for 10 min at room temperature before the absorbance at 550 nm was measured. Cholesterol reduction (%) was calculated as: 1 – (absorbance of culture supernatant 550/absorbance of control 550) \(\:\times\:\) 100 Exopolysaccharide (EPS) producing ability of Lactobacillus isolates Pure cultures of the samples were evaluated according to the method of Abouloifa et al. ( 37 ), for the ability of Lactobacillus species to produce EPS. Freshly cultivated isolates were streaked on ruthenium red milk agar plates containing 10% (w/v) skim milk (Merck, SA), 1% (w/v) sucrose (Sigma, SA), 0.08% (w/v) ruthenium red (Sigma, SA), and 1.5% (w/v) agar (Merck, SA). After anaerobic incubation for 48 h at 37°C, white colonies indicated a positive result for EPS production, whereas pink colonies indicated a negative outcome. 2,2-Diphenyl-1-picrylhydrazyl (DPPH) scavenging activity The method proposed by Kim et al. ( 38 ) was used with slight modifications to determine the DPPH radical scavenging ability of the isolates. A DPPH radical solution was prepared by dissolving 4 mg of DPPH in 100 ml of methanol in a glass tube wrapped in aluminum foil. Two milliliter of DPPH solution was added to 1 ml of each bacterial suspension (OD600 nm = 1.0) and mixed thoroughly. The resulting mixture was kept in the dark at room temperature for 30 min. A mixture of 2 ml of DPPH and 2 ml of methanol was used as a negative control, while ascorbic acid (10 mg/ml) (Merck, SA) served as the positive control. The assay was performed in triplicate, and the absorbance (OD) of the resulting solution was measured at 517 nm. The scavenging effect (%) = (Ac–As)/Ac \(\:\times\:\) 100, where As is the absorbance of the test sample, and Ac is the absorbance of the control sample. Antimicrobial activity against pathogenic bacteria The agar well diffusion method described by Sakandar et al.( 39 ) was used to examine the antimicrobial activity of Lactobacillus isolates against pathogenic bacteria. The test pathogens used in this study were Staphylococcus aureus ( S. aureus ) ATCC 25923, Escherichia coli ( E. coli ) ATCC 25922, Listeria monocytogenes ( L. monocytogenes ) ATCC 15313, and Bacillus cereus ( B. cereus ) ATCC 11778 (Anatech, SA). Lacticaseibacillus rhamnosus GG served as a probiotic reference strain. The cell-free supernatant (CFS) of each isolate was prepared by incubating 10 ml of MRS broth at pH 7.2 overnight at 37°C, followed by centrifugation at 10,000 rpm for 5 min at 4°C. The CFS was collected using a sterile syringe with a needle. The CFS was then sterilized using a 0.22 µm filter (Anatech, SA). Each pathogenic strain was grown overnight at 37°C in nutrient broth (Sigma, SA) and adjusted to an OD of 0.25 at 600 nm. Approximately 0.1 ml of each pathogen cell suspension was spread on Muller-Hinton agar (MHA) (Merck, SA) using a sterile cotton swab. The agar plates were allowed to dry for 1–2 min before wells were created on the MHA plate using the SBPT. The CFS of the isolates were pipetted into each agar well, and the plate was incubated at 37°C for 16–24 h. Clear zones around each agar well were measured in millimeter (mm). Cell auto - aggregation Four ml of each bacterial suspension (OD600 nm = 0.25) was vortexed for 10 s, after which the suspension was incubated at 37°C for 2 and 4 h. An aliquot of 1 ml of the upper suspension was taken using a sterile Pasteur pipette, and its absorbance was determined at an OD of 600 nm. Auto-aggregation (%) = [(1– (A t /A 0 )] \(\:\times\:\) 100, where A t is the absorbance at time t = 2 or 4 h, while A o is the absorbance at time t = 0 h ( 40 ). Cell surface hydrophobicity The cell surface hydrophobicity was determined using the method described by Barache et al. ( 40 ). The initial absorbance of the bacterial cell suspension (A 0 ) at 600 nm was recorded. One ml of xylene was added to three ml of each cell suspension (OD600 nm = 0.25), and mixed by vortexing for 2 min. The bacterial suspension was incubated at 37°C for 1 h to allow phase separation. The lower aqueous phase was carefully removed, and its absorbance (A t ) at OD 600nm was measured. Cell surface hydrophobicity (%) = [(1– (A t /A 0 )] × 100, where A t is the absorbance of the aqueous phase and A 0 is the absorbance of the initial bacterial suspension. Safety evaluation of Lactobacillus isolates Antibiotic susceptibility The disk diffusion method described by Mohammad et al. ( 41 ) was used in this study to assess the antibiotic susceptibility of the isolates. Approximately 0.1 ml of each bacterial cell suspension (OD600 nm = 0.25) was spread evenly on the surface of the MRS agar plate and allowed to dry for approximately 8–15 min. Subsequently, different antibiotic discs (Separations, SA) containing ampicillin (10 µg), erythromycin (15 µg), chloramphenicol (30 µg), tetracycline (30 µg), gentamycin (10 µg), streptomycin (10 µg), vancomycin (30 µg) penicillin G (10 µg) and kanamycin (30 µg) were placed on the surface of the inoculated agar plates aseptically with sterile forceps. After incubation at 37°C for 24 h, the diameters of the inhibition zones were measured using a ruler in mm. The inhibition zones were categorized as sensitive (≥ 21 mm), intermediate (16–20 mm), or resistant (≤ 15 mm), as detailed by Reuben et al. ( 42 ). Haemolytic activity Each isolate was streaked onto Columbia agar (Anatech, SA) plates supplemented with 5% (w/v) sterile sheep blood. After incubation at 37°C for 24–48 h, the plates were examined for signs of haemolysis: β-haemolysis (clear zones around colonies), α-haemolysis (green-hued zones around colonies), or ϒ-haemolysis (no visible zones around colonies) ( 29 ). Deoxyribonuclease (DNase) activity Isolates were tested for DNase enzyme production by streaking each culture (OD600 nm = 0.25) onto DNase agar (Separations, SA) plates. After incubation at 37°C for 24–48 h, the plates were flooded with 1 M HCl. A clear zone around the colonies was considered positive for DNase production ( 37 ). S. aureus ATCC 25923 was used as a positive control. Cell adhesion assay The human colon adenocarcinoma cell line (Caco-2 cells) (CELLONEX, SA) was kindly provided by Dr. Wale Oladipo (Department of Life Sciences, University of South Africa). Caco-2 cells were grown in a humidified incubator of 5% CO 2 at 37°C in Dulbecco’s modified Eagle medium (DMEM) (Sigma-Aldrich, SA) supplemented with 10% inactivated fetal bovine serum (Sigma-Aldrich, SA), 100 units/ml penicillin (Thermo Fisher Scientific, SA), and 0.1 mg/ml streptomycin (Thermo Fisher Scientific, SA). To achieve 97% confluency, the cells were harvested and counted using a hemocytometer. The Caco-2 cells were adjusted to a concentration of 1.5 × 10 5 cells/ml by counting the cells that detached from the flask. The Caco-2 cells were subsequently seeded at 500 µl per well in a 36-well culture plate and incubated in a humidified 5% CO 2 incubator for 24 h at 37°C. The culture medium was changed every 2 days for 15 days to obtain a monolayer of differentiated Caco-2 cells. The adhesive activity of the isolates was examined using the method described by Fonseca et al. ( 43 ). Lactobacillus isolates were cultured in MRS broth at 37°C for 18 h. The grown bacterial cultures were then centrifuged at 8000 \(\:\times\:\) g for 10 min, and the cell pellets were collected, washed twice with sterile PBS, and then suspended in DMEM without antibiotics before being standardized to an OD600 nm of 1.0. Next, 1 ml of each bacterial suspension was added to the monolayer of differentiated Caco-2 cells in the wells of separate 36-well culture plates and incubated for 90 min at 37°C in a 5% CO 2 humidified incubator. After incubation, the cells in the wells were washed three times with 1 ml of prewarmed PBS to remove any unbound Lactobacillus cells. Thereafter, the bounded Caco-2 cells and bacterial cells were detached from each well by adding 1 ml of Triton-X solution (0.1% v/v in PBS) and incubating for 10 min at 37°C. After incubation, the Caco-2 cells and Lactobacillus suspensions were harvested and centrifuged at 9000 \(\:\times\:\) g for 10 min at 4°C. The resulting pellet was washed twice with prewarmed PBS before being resuspended in PBS. Finally, 100 \(\:\mu\:\) l of appropriate serial dilutions of the Caco-2 cell/bacterial cell suspension were spread on MRS agar plates and incubated at 37°C for 48 h before counting. Adhesion ability (%) \(\:\frac{\text{I}\text{n}\text{i}\text{t}\text{i}\text{a}\text{l}\:\text{c}\text{o}\text{u}\text{n}\text{t}\text{s}\:\text{o}\text{f}\:\text{b}\text{a}\text{c}\text{t}\text{e}\text{r}\text{i}\text{a}\text{l}\:\text{s}\text{e}\text{e}\text{d}\text{e}\text{d}\:}{\text{T}\text{h}\text{e}\:\text{c}\text{o}\text{u}\text{n}\text{t}\text{s}\:\text{a}\text{f}\text{t}\text{e}\text{r}\:\text{t}\text{h}\text{e}\:\text{w}\text{a}\text{s}\text{h}\text{i}\text{n}\text{g}\:\text{s}\text{t}\text{e}\text{p}\text{s}}\) \(\:\times\:\) 100 Lacticaseibacillus rhamnosus GG was used as a positive control. Cumulative probiotic potential (CPP) of Lactobacillus isolates The probiotic potential of the 11 isolates from this study was assessed based on their cumulative probiotic score. The sum of acid tolerance, bile salt tolerance, auto-aggregation ability, cell surface hydrophobicity, antimicrobial activity, DNase activity, haemolytic activity, and antibiotic susceptibility is the CPP ( 36 ). Probiotic Potential (%) = \(\:\frac{\text{O}\text{b}\text{s}\text{e}\text{r}\text{v}\text{e}\text{d}\:\text{s}\text{c}\text{o}\text{r}\text{e}}{\text{M}\text{a}\text{x}\text{i}\text{m}\text{u}\text{m}\:\text{s}\text{c}\text{o}\text{r}\text{e}}\) \(\:\times\:\) 100 ( 36 ) Statistical analysis All experiments were performed in triplicate, and descriptive statistics are presented as the mean ± standard deviation (SD). One-way analysis of variance (ANOVA) was employed, and Duncan’s multiple range test was used to compare significant differences between the mean values of acid tolerance, bile salt tolerance, auto-aggregation ability, cell surface hydrophobicity, cholesterol-lowering ability, antibiotic susceptibility, antioxidant activity, and cell adhesion. The Statistical Package for the Social Sciences (SPSS) program, version 29, was used for all analyses, and significance was considered at P ≤ 0.05. Results Identification and phylogenetic analysis of Lactobacillus isolates Lactobacillus species were identified in all the sources; a total of 18 presumptive Lactobacillus isolates were obtained from the samples and characterized as catalase-negative, non-motile, Gram-positive, and rod-shaped. These Lactobacillus isolates were identified as: Lactobacillus johnsonii (n=7; 38.9%), Latilactobacillus curvatus (n=4; 22.2%), Lactiplantibacillus plantarum (n=4; 22.2%), Limosilactobacillus reuteri (n=2; 11.1%), and Latilactobacillus sakei (n=1; 5.6%) ( Table 1 & Table 2 ). As presented in Table 1 , Lactobacillus johnsonii was the dominant Lactobacillus species isolated from the chicken gut, while Lactiplantibacillus plantarum was the prevalent species identified in vegetable samples. On the other hand, Latilactobacillus curvatus was the only Lactobacillus strain isolated from the banana samples. Table 1 : Lactobacillus species identified from chicken gut, tomatoes, cucumber and banana Table 2 : Identification of Lactobacillus species isolated from chicken gut, tomato, cucumber, and banana using 16S rRNA Gene Sequencing The results depicted in Figure 1 revealed that the Lactobacillus strainsare present in three distinct clades. One clade consisted of the Latilactobacillus curvatus strains Chcg (chicken gut) and Pb2 (banana), establishing their close genetic relationship with Latilactobacillus curvatus strains P1-S-OK326292 and 1224 MT573677 and Latilactobacillus sakei strain 6699 MT463883. These findings demonstrated the possible evolution of these isolates from a similar ancestor. The second clade consisted of Lactiplantibacillus plantarum strains To3a (tomato), To3d (tomato), and Latilactobacillus curvatus strain Chcb (chicken gut), having a close relationship with Latilactobacillus sakei strain HBUAS5613 and Latilactobacillus curvatus strain TM332D OM265415, clearly indicating that they share a common ancestor. Lactiplantibacillus plantarum strain (To3d, tomato) clustered closely with Lactiplantibacillus plantarum strain LP 212 OMO 38099, indicating a shared ancestry. In contrast, Lactiplantibacillus plantarum strain (To3a, tomato) was differently clustered along Lactiplantibacillus plantarum strain LP 212 OMO 38099, Latilactobacillus sakei strain HBUA S56133 MT211361 and Latilactobacillus curvatus strain TMPC 3332D OMO 265415, implying that they may not share a similar ancestor. On the other hand, Lactiplantibacillus plantarum strains (To3b, tomatoes and Cu3e, cucumber), Latilactobacillus curvatus strain (Cu2f, cucumber) and Latilactobacillus sakei strain (Mcg, chicken gut) were clustered within the third clade. These strains formed a distinct sub-clade, indicating that they share a different common ancestor from the other isolates. Latilactobacillus sakei strain (Mcg, chicken gut) shares a common ancestor with Latilactobacillus sakei strains 2751 MT611760, 2178 MT604678 and SDCM 1005 MW548739. As shown in Figure 2 , the phylogenetic tree indicates that Lactobacillus johnsonii strain (Chcx4a, chicken gut) is closely related to Lactobacillus johnsonii strain 20678 MW866875 in one clade but also connects to multiple branches of subclades consisting of Lactobacillus johnsonii strains (Chcx2, Chcx3a, and Mcxb, chicken gut). Lactobacillus johnsonii strains Mcyc, Mcxc and Mcya (chicken gut) were closely related to Lactobacillus johnsonii strains MH33 FJ5422 and 33 PP728187, indicating a shared ancestry. In contrast, Limosilolactobacillus reuteri strains Chcx3a and Mcyc (chicken gut) exhibited a distant phylogenetic relationship with the other strains, suggesting that they do not share a common ancestor with the other strains. Acid tolerance The isolates Lactiplantibacillus plantarum (To3b and To3d, tomato), Limosilactobacillus reuteri (Chx3a and Mcyc, chicken gut), Lactobacillus johnsonii (Chx3b, Mcxb, and Mcxc, chicken gut), and Latilactobacillus curvatus (Cu2f, cucumber), displayed acid tolerance rates that were not significantly (p >0.05) different between pH 3.0 and pH 2.5. Whereas a lower survival rate significantly (p>0.05) different from the rest of the isolates was demonstrated by isolates Latilactobacillus curvatus (Cu2f, cucumber) and Lactobacillus johnsonii (Mcxb and Mcxc, chicken gut) at pH 2.5 ( Table 3 ). The isolates Lactiplantibacillus plantarum (To3b and To3d, tomato), Lactiplantibacillus plantarum (Cu3e, cucumber), Latilactobacillus curvatus (Pb2, banana), Limosilactobacillus reuteri (Mcyc, chicken gut), Limosilactobacillus reuteri (Chx3a, chicken gut), and Lacticaseibacillus rhamnosus GGexhibited significantly (p ≤ 0.05) higher survival rates at pH 3, with no significant differences (p>0.05) observed among isolates Lactiplantibacillus plantarum (To3a, tomato), Limosilactobacillus reuteri (Mcyc, chicken gut), and Lacticaseibacillus rhamnosus GG. Meanwhile, isolates Lactilactobacillus curvatus (Cu2f, cucumber) and Lactobacillus johnsonii (Mcxc, chicken gut) exhibited relatively low survival rates of 53.79% and 65.65%, respectively, which were significantly different (p > 0.05) from the rates of the other isolates at pH 3( Table 3 ). Out of the 18 isolates analysed above, seven, Latilactobacillus sakei (Mcg, chicken gut), Lactobacillus johnsonii (Chcx4a, Chcx2, Mcya, and Mcyb, chicken gut), Latilactobacillus curvatus (Chcb and Chcg, chicken gut), exhibited a survival rate between 7% to 20% (data not shown) at pH 3.0 and hence were excluded from the probiotic due to their poor acidic tolerance, resulting in the total of 11 Lactobacillus isolates used for further analysis. Table 3: The acid tolerance level of Lactobacillus isolates after exposure to pH 2.5 and pH 3.0 for 3h Bile salt tolerance The isolates Lactiplantibacillus plantarum (To3b and To3d, tomato), Limosilactobacillus reuteri (Chx3a and Mcyc, chicken gut), Lactobacillus johnsonii (Chx3b, Mcxb, and Mcxc, chicken gut), Lactiplantibacillus plantarum (Cu3e, cucumber) and Latilactobacillus curvatus (Pb2, banana) exhibited bile salt tolerance that was not significantly (p > 0.05) different between 0.3% bile salt and 0.5% bile salt. On the other hand, the other isolates, including the reference strain, had a significantly (p ≤ 0.05) lower cell survival rate at 0.5% bile salt ( Table 4 ). Table 4: The bile salt tolerance level of Lactobacillus isolates after exposure to 0.3% and 0.5% bile salt for 4 h BSH activity Three isolates, Lactiplantibacillus plantarum (To3a, tomato), Limosilactobacillus reuteri (Chcx3a, chicken gut) and Lactiplantibacillus plantarum (Cu3e, cucumber) revealed the largest precipitation zones 10–15 mm (++) compared with the reference strain and other isolates. However, Lactobacillus johnsonii (Mcxc, chicken gut) showed no precipitation zone in this study ( Table 5 ). Table 5: The BSH activity, EPS production ability, and cholesterol-lowering ability of Lactobacillus isolates The EPS producing ability Most isolates, Lactiplantibacillus plantarum (To3a, To3b, To3d, tomato), Lactiplantibacillus plantarum (Cu3e, cucumber), Limosilactobacillus reuteri (Chx3a, chicken gut), Latilactobacillus curvatus (Cu2f, cucumber), and Lactobacillus johnsonii (Mcxb, chicken gut), showed the ability to produce EPS, similar to the reference strain. Conversely, Lactobacillus johnsonii (Chx3b and Mcxc, chicken gut), Latilactobacillus curvatus (Pb2, banana), and Limosilactobacillus reuteri (Mcyc, chicken gut) did not exhibit the ability to produce EPS ( Table 5 ). Cholesterol-lowering ability of Lactobacillus isolates The tested isolates exhibited varying degrees of cholesterol-reducing abilities, ranging from 13.06% to 57.64% ( Table 5 ). The isolates Lactiplantibacillus plantarum (To3a and To3d, tomato), Limosilactobacillus reuteri (Chx3a, chicken gut), and Lactiplantibacillus plantarum (Cu3e, cucumber) exhibited significantly (p ≤0.05) higher cholesterol-lowering abilities than the reference strain (41.81%). Isolates To3d and Chx3a exhibited a cholesterol-lowering ability of more than 50%. DPPH scavenging activity The tested isolates exhibited varying degrees of DPPH scavenging activity, ranging from 6.81% to 24.45% compared with 35.63% from ascorbic acid. The DPPH scavenging activity of Lactiplantibacillus plantarum (To3d, tomato) (24.45%) was significantly (p ≤ 0.05) higher than that of the reference strain (23.20%). In contrast, that of Limosilactobacillus reuteri (Mcyc, chicken gut) (20.90%) was not significantly different (P>0.05) from that of the reference strain ( Table 6 ). Table 6: DPPH scavenging activity (%) of Lactobacillus isolates A ntimicrobial activity against pathogenic bacteria The tested Lactobacillus isolates exhibited varying degrees of antimicrobial activity against selected pathogens. The isolates Lactiplantibacillus plantarum (To3a, To3b, and To3d, tomato) demonstrated antimicrobial activity similar to that of the reference strain, with an inhibition zone of 10 to 15 mm (++) or more against E. coli , S. aureus , B. cereus , and L. monocytogenes ( Table 7 ). Table 7: Antimicrobial activity of CFS of Lactobacillus against pathogenic organisms Cell auto-aggregation The auto-aggregation (%) of selected Lactobacillus isolates generally increased after incubation from 2 to 4 h, as shown in Table 8 . Compared with the reference strain, the isolates Lactiplantibacillus plantarum (To3a, To3b and To3d, tomato), Limosilactobacillus reuteri (Chx3a and Mcyc, chicken gut), Lactobacillus johnsonii (Chx3b, chicken gut), and Latilactobacillus curvatus (Cu2f, cucumber), displayed a significant (p ≤ 0.05) increase in auto-aggregation (%) after incubation from 2 to 4 h. Similarly, the auto-aggregation (%) of these isolates was significantly (p ≤ 0.05) higher than that of the reference strain after incubation for 2 and 4 h, respectively. The other strains displayed auto-aggregation (%) between 18.00% and 30.67% after 4 h incubation, which was significantly different (p≤ 0.05) from that demonstrated by the reference strain at 24.67%. Table 8: Theauto-aggregation ability of Lactobacillus isolates after 2 h and 4 h incubation Cell surface hydrophobicity The Lactobacillus isolates exhibited varying degrees of cell surface hydrophobicity ( Table 9 ). The Latilactobacillus curvatus (Cu2f, cucumber), Lactiplantibacillus plantarum (Cu3e, cucumber), and Limosilactobacillus reuteri (Mcyc, chicken gut) showed cell surface hydrophobicity values significantly (p ≤ 0.05) higher than those of Lacticaseibacillus rhamnosus GG,whereas Lactobacillus johnsonii (Chx3b, chicken gut) exhibited cell surface hydrophobicity (%) not significantly different (p>0.05) from those of the reference strain. The remaining isolates presented a cell surface hydrophobicity (%) between 0.00% and 46.67% after 1 h of incubation, which was significantly different (p ≤ 0.05) from that of the reference strain (53.33%). Table 9: Cell surface hydrophobicity ability of Lactobacillus isolates after 1 hr incubation Antibiotic susceptibility The Lactobacillus isolates showed varying antibiotic susceptibility profiles. All the isolates were susceptible to penicillin G (10 μg), chloramphenicol (30 μg), erythromycin (15 μg) and ampicillin (10 μg). On the contrary, all the isolates were resistant to kanamycin (30 μg), gentamycin (10 μg), and streptomycin (10 μg) ( Table 10 ). Table 10: Antibiotic susceptibility profile of Lactobacillus isolates using the disk diffusion method Haemolysis and DNase activity None of the Lactobacillus isolates in this study exhibited haemolysis or DNase activity ( Table 11 ). Table 11: Bloodhaemolysis and DNase activity of Lactobacillus isolates Cell adhesion assay Lactobacillus johnsonii (Chx3b, chicken gut) and Lactiplantibacillus plantarum (To3d, tomato) showed Caco-2 cell adhesion capacities (15.29% and 12.87% respectively) higher than that of the reference strain (11.40%), whereas the isolate Limosilactobacillus reuteri (Mcyc, chicken gut) presented a slightly lower adhesion capacity of 10.53%. In contrast, isolates Latilactobacillus curvatus (Pb2, banana) and Lactiplantibacillus plantarum (Cu3e, cucumber) demonstrated the lowest adhesion capacities at 2.60% and 3.58%, respectively ( Table 12 ). Table 12: Adhesion capacity of Lactobacillus isolates to Caco-2 epithelial cells CPP of Lactobacillus isolates The CPP of the eleven isolates, as presented in Table 13 , are 50% for isolate Latilactobacillus curvatus (Pb2, banana), and Lactobacillus johnsonii (Mcxb, chicken gut); 62.50% for isolate Limosilactobacillus reuteri (Chx3a, chicken gut), Lactiplantibacillus plantarum (Cu3e, cucumber), and Lactobacillus johnsonii (Mcxc, chicken gut); 75% for isolate Lactiplantibacillus plantarum (To3b, tomato), Lactobacillus johnsonii (Chx3b, chicken gut), and Limosilactobacillus reuteri (Mcyc, chicken gut), and 87.50% for the remaining three isolates: Lactiplantibacillus plantarum (To3a and To3d, tomato) and Latilactobacillus curvatus (Cu2f, cucumber). Table 13: Cumulative probiotic potential of Lactobacillus isolates Discussions Identification of Lactobacillus strains Lactobacillus species were identified in all the sample types, possibly because they are widely distributed in various ecological niches and are adapted to survive naturally in nutrient-rich and moist environments, which aids their growth and colonization ( 44 ). Among several species of Lactobacillus strains, Limosilactobacillus reuteri and Lactobacillus johnsonii have been observed to be among the most detected microbes in poultry GIT ( 45 ). Additionally, Junnarkar et al. ( 46 ) reported in their study that the genus Lactobacillus was dominant in fresh vegetable samples (cabbage, cauliflower, cluster bean, French beans, gherkins, bitter gourd, rigid gourd, fenugreek, and tomatoes). In particular, Lactiplantibacillus plantarum was one of the most prevalent bacterial species found in tomatoes, fermented cabbage, and olives ( 47 ). Bacterial strains derived from plant-based matrices, such as fruits and vegetables, and the animal gut, possess notable genetic and functional diversity that the food industry may leverage to foster innovative probiotic applications ( 48 ). Lactobacillus johnsonii is a resident microflora of the poultry GIT that particularly adheres to the epithelium of the duodenum and jejunum ( 49 , 50 ). Lactobacillus johnsonii was the most prevalent Lactobacillus species in the chicken gut samples, possibly because of external factors, such as chicken feed, antibiotic treatment, and invading organisms ( 50 ). Furthermore, the level of stress, environmental conditions, and animal health may also play a role in the differences in the microbial composition of the chicken gut ( 51 ). Similar to our findings, a relatively high occurrence of Lactobacillus johnsonii (strains AER105 and AER25) isolated from the ilea of five individual chickens ( 52 ), and Lactobacillus johnsonii ; 21% in chicken mucosa of the ileum (M-IM), 15% in the cecal lumen (M-CL) and 11% in the mucosa of ceca (M-CM) ( 53 ) was established in their study. However, previous studies have reported that Lactobacillus johnsonii isolated from the GIT of chickens is prevalent in 23.3% ( 54 ); 10% ( 55 ), 7.69% ( 18 ), and 5% ( 51 ). ( 56 ) mentioned that the type of chicken breed can influence their gut microbiota, for example, laying hens have different microbial communities than the broiler breeds used in this study. This is owing to their physiological and genetic variations ( 57 ). In addition, feed formulations, including the use of probiotics, protein, and fibre in the chicken diet, can favourably impact the overall microbial population ( 56 , 58 ). According to Bindari & Gerber ( 59 ), higher levels of Lactobacillus species were noted throughout the GIT of chickens fed whole wheat than in chickens fed grounded wheat pellets. Housing systems, such as the use of cages, barns and free-range systems ( 57 ), and the geographical location of the chickens (high altitude or low altitude regions) may also reshape the microbiota landscape and colonization ( 56 ). The prevalence of Lactobacillus johnsonii represents an important tool that could be utilized as probiotics in poultry feed for improving the gut microbial structure, weight gain, enhancing feed conversion ratio, and overall performance, especially in boilers ( 60 ). The next most dominant microbe was Latilactobacillus curvatus , which was isolated from the chicken gut, banana and cucumber. Latilactobacillus curvatus is commonly associated with lactic acid fermentation and can be isolated from several environments, including fermented meat, fermented vegetables, dairy products, human GIT, and other niches ( 61 ). Latilactobacillus curvatus is mostly associated with fermented meat and poultry products ( 62 ), although it may be isolated from foods of vegetable origin as well as the GIT of animals that feed on plants or cereals, including chickens ( 63 ). Latilactobacillus curvatus can inhibit the growth of foodborne pathogens in fermented food products via the production of bacteriocin ( 61 ). Besides, they play an essential role in the preservation and flavour development of fermented dairy foods via lactic acid production and the development of desirable flavours in meat products ( 64 ). Additionally, the incorporation of Latilactobacillus curvatus strains such as SMFM2016-NK in fermented milk may lessen periodontitis and regulate oral and gut microbial composition ( 65 ). Lactiplantibacillus plantarum sourced from tomatoes and cucumbers was the dominant strain in this study. This microbe is predominantly found in fermented food, including cucumbers and tomatoes ( 66 , 67 ), and plant surfaces ( 68 ), and it is one of the most versatile and metabolically diverse organisms commonly involved in vegetable fermentation of products such as kefir and sauerkraut ( 69 ). A previous study revealed that they can enhance their genome for growth in an environment rich in plant carbohydrates, owing to the incidence of genomic islands sheltering mosaic modules or cassettes of carbohydrate utilization genes, which may have been acquired through horizontal gene transfer ( 70 ). Borjihan et al. ( 71 ) mentioned that Lactiplantibacillus plantarum has a distinct adaptability to the vegetable environment, which may be attributed to its stress response pathways and large metabolic capacity. Additionally, the ability of this bacterial species to produce antimicrobial compounds and biofilm capacity are attributes that enable it to survive and dominate in such an environment ( 48 ). Lactiplantibacillus plantarum was reported to be prevalent in 56.82% of raw tomatoes ( 72 ), 34.6% of vegetable residues and silage fermentation ( 73 ), 55.55% ( 74 ) and 57.69% in Chinese fermented vegetables ( 75 ). On the contrary, 19.2% Lactiplantibacillus plantarum was dominant in plant-based samples ( 48 ), 11.11% in fresh vegetables ( 76 ), and 3.89% in cabbage and tomatoes ( 77 ). The probiotic Lactiplantibacillus plantarum strain plays a pivotal role in the gut microbial regulation and reduction in total body fat and body weight gain, as demonstrated in a high-fat-diet obese mice trial ( 78 ). In addition, this bacterium may be a promising probiotic candidate for improving fermentation-based food products ( 48 ). On the other hand, Latilactobacillus curvatus was the only Lactobacillus species isolated from the banana fruit samples. This fermentation bacterial strain is usually found in the fruit matrix and has remarkable fermentation properties, which could result in improved preservation and nutritional qualities ( 79 ). The reason for this may be that Latilactobacillus curvatus bacteria possess numerous genes that encode metabolic pathways for carbohydrate utilization, such as starch, which acts as an ideal substrate that encourages the colonization and growth of this microbe ( 62 ) in bananas. Conversely, 35% Latilactobacillus curvatus isolates were derived from Italian red and yellow pepper fruits ( 80 ), 38.19% from Saudi chicken ceca samples ( 81 ), and 20% from fermented Chinese sausage ( 82 ). Soil health and quality, for example, the planting of banana with high-fertility soil, may contribute to a robust bacterial community composition ( 83 ). In addition, Kaushal et al. ( 84 ) mentioned that differences in climatic conditions, such as temperature, rainfall, and geographical characteristics of banana-producing areas, may significantly contribute to variations in the microbial diversity of bananas. Latilactobacillus curvatus isolate derived from bananas may hold a significant promise in the health industry due to its potential probiotic impact in modulating immune response, reducing colitis and improving gut barrier functions ( 85 ). In the food industry, they may serve as a bio-preservative agent by producing lactic acid, which is useful for the preservation and flavour development of fermented dairy food ( 64 ). Acid tolerance The acid tolerance of selected Lactobacillus isolates generally increased at pH 3.0, as shown in Table 3 . The acidic pH of the stomach (2.5–3.5) results in the formation of a natural barrier against external bacteria ( 86 ), requiring Lactobacillus species to tolerate such low pH conditions to survive and remain active during GIT transit, particularly over the estimated 3 h period it spends in the stomach during food passage ( 87 ). The isolates, Lactiplantibacillus plantarum (To3a, To3d, tomatoes), Lactiplantibacillus plantarum (Cu3e, cucumber), Latilactobacillus curvatus (Pb2, banana), Limosilactobacillus reuteri (Mcyc, Chx3a, chicken gut), and Lacticaseibacillus rhamnosus GG, exhibited significantly (p ≤ 0.05) higher survival rates at pH 3. Our results highlighted that these Lactobacillus species are highly acid-tolerant strains, and that their ability to survive in acidic medium differs among different species and sources. Most Lactobacillus species can tolerate acidic environments (pH 2–3) because of their remarkable ability to metabolize sugars to generate lactic acid ( 88 ). Furthermore, Hojjati et al. ( 89 ) and Barzegar et al. ( 90 ) reported that Lactobacillus can develop distinct mechanisms for survival in low acidic conditions, the most important of which is F 0 F 1 -ATPase. This mechanism shields the bacterial cell from acidic exposure by transferring the protons out of the cell membrane and thus maintaining the intracellular pH ( 36 ). Other mechanisms by which Lactobacillus species tolerate high acidity include H + pumping to an acidic environment, acid end-product efflux, the synthesis of alkali products to neutralize acid produced during extracellular metabolism, the repair of macromolecules, biofilm formation, and pre-adaptation and cross-protection ( 91 ). Additionally, the EPS produced by Lactobacillus forms a layer in a low pH environment that limits the access of exogenous acids to the bacterial cells due to anions bound to EPS as a phosphate group ( 92 ). In a separate study, Lactiplantibacillus plantarum strains O19, O20, O21, O23, and K1 and Lactobacillus johnsonii strain K4, which were isolated from traditional fermented cabbage and cucumber, also showed a high tolerance rate > 90% to pH 2.5 and pH 3.5 ( 27 ). Ahire et al. ( 93 ) found that Lactiplantibacillus plantarum UBLP40 isolated from indigenous fermented food had a 93% survival rate at pH 3.0 after 3 h of incubation. Lactiplantibacillus plantarum strains sourced from Italian food products showed high tolerance to pH 3.0 and 2.5, presenting a survival rate ranging from 60.69% to 100.08% ( 94 ). In another study, Rajoka et al. ( 95 ) reported that six Limosilactobacillus reuteri strains and one Lactobacillus johnsonii strain sourced from the chicken GIT demonstrated a high survival rate greater than 80% at pH 3.0 after 3h of incubation, whereas five out of seven Lactiplantibacillus plantarum strains showed a survival rate > 70% at pH 3.0 after 5 h incubation ( 37 ). High resistance to pH 3.0 conditions was also observed for 25 Limosilactobacillus reuteri strains, with bacterial growth rates ranging from 60.27% to 100.48% after 24 h of exposure ( 96 ). Meanwhile, the survival ability of Lactiplantibacillus plantarum strains (AF(PIF), P8014, M8, LFS7 and a7) isolated from different sources at pH 2.5 was reported to be 98.85% to 100% after 4 h of exposure ( 34 ), while Tokatl et al.( 95 ) reported that at pH 2.5, twenty-one Lactiplantibacillus plantarum strains sourced from traditional pickles showed survival ability of 35% – 85% after 4 h of exposure. These results highlight the potential role of these strains in the development of probiotic supplements without a coating ( 97 ), and in maintaining cell viability, especially in acidic food systems ( 31 ) like fermented fruits and vegetables. The survival rates of the other isolates did not differ significantly at pH 2.5 and pH 3. This is possibly due to the strain-specific nature of these strains and their remarkable resilience in an acidic environment ( 93 ). The differences observed in our results regarding the acidic tolerance rates of lactobacilli isolates may be attributed to the bacterial source and the duration of acidic exposure ( 93 ). Furthermore, acid tolerance variation in LAB has been connected to the dissimilarity in H + -ATPase activity induction, leading to the removal of protons (H + ), alkalinization of the outside environment, and alterations in the cell envelope composition ( 98 ). Our findings corroborate the report by Zielińska et al. ( 27 ), who found that at pH 2.5 and pH 3.5, there were no significant changes in the survival rates of Lactiplantibacillus plantarum (O19, O20, O21, O23, and K1) and Lactobacillus johnsonii (K4) isolates in their study, as all exhibited a survival rate greater than 90%. Manzoor & Tayyeb( 14 ) revealed that all Lactiplantibacillus plantarum isolates maintained a constant viable cell count at pH 3 after 4 h of exposure. In addition, Lactiplantibacillus plantarum SW03 and SW07 were not significantly different, and their survival rate was above 90% when exposed to pH 2.5 and 3.0 after 3hr incubation ( 29 ). Likewise, no significant differences were recorded between Limosilactobacillus reuteri TF-7 and Lacticaseibacillus rhamnosus GG after exposure to pH 2.0, 3.0, and 4.0 ( 99 ). In contrast, an Italian study conducted by Turchi et al. ( 94 ) reported high growth rates, ranging from 96.98% to 102.87% at pH 3.0, were observed by all 37 Lactiplantibacillus plantarum strains, while at pH 2.5, survival rates ranging from 60.69% to 100.08% were recorded. Furthermore, Mulaw et al. ( 100 ) reported that Lactiplantibacillus plantarum EO52 and TO35 were significantly different at pH 2.5 and 3.0, after exposure for 6 h. The isolates Lactiplantibacillus plantarum (To3a, tomato) and Limosilactobacillus reuteri (Mcyc, chicken gut) exhibited survival rates of 97% and 98.1%, respectively, at pH 3, which were higher than those of Lacticaseibacillus rhamnosus GG, with a survival rate of 93%. Lactiplantibacillus plantarum and various homofermentative lactobacilli can balance their intracellular redox state using an extracellular electron transfer mechanism ( 101 , 102 ). Lactobacillus species of vegetable origin, such as tomato, are generally more pH-tolerant ( 75 ). Zhang et al. ( 103 ) pointed out that the resilience of Lactiplantibacillus plantarum to low acidic concentration of pH 3.0 was positively linked with the production of EPS, as the low pH tolerance of these bacteria can be improved by adjusting their plasma membrane fatty acid composition to decrease fluidity, increasing the proton pump activity to maintain the intracellular pH balance. According to Nguyen et al. ( 92 ), an increase in biofilm formation in EPS-producing Limosilactobacillus reuteri strains was noted, compared with a decrease in the formation of biofilm in EPS-producing Lacticaseibacillus rhamnosus GG under low pH conditions. Additionally, species like Limosilactobacillus reuteri naturally inhabit the human or animal small intestine; hence, they are better genetically adapted to the low pH conditions of the gut than those of food origin ( 104 , 105 ). Owing to their enhanced host adaptation, greater cell viability, and safety, probiotic strains isolated from their natural hosts, such as the animal gut, are chosen over those derived from other sources ( 106 ). Our results agree with those obtained from an earlier study, where Lactiplantibacillus plantarum DUR5 and DUR8 demonstrated a higher survival rate of 87.61% and 90.24%, respectively, whereas Lacticaseibacillus rhamnosus GG exhibited 80.61% resistance to pH 3.0 ( 107 ). A study by Jomehzadeh et al. ( 108 ) noted that Lactiplantibacillus plantarum N12 to N20 showed survival rates ranging from 60% to 96% compared with 58% demonstrated by Lacticaseibacillus rhamnosus GG at pH 2.5. In addition, Lactiplantibacillus plantarum LpJ1 and LpJ5 demonstrated significantly higher viable cell counts of 7.93 CFU/ml and 7.93 CFU/ml, respectively, than did Lacticaseibacillus rhamnosus GG (6.00 CFU/ml) at pH 2.5 after 180 min incubation ( 109 ). In contrast, a higher survival rate of 86.48% was demonstrated by Lacticaseibacillus rhamnosus GG exposed to gastric juice at pH 2.5 for 3 h, compared with Lactiplantibacillus plantarum LN-3-1, which had a 38.06% survival rate ( 110 ). Lactiplantibacillus plantarum strain C06 exhibited tolerance levels greater than 90% when exposed to acidic conditions at pH 3 ( 111 ), but this value is less than the 110% survival rate achieved by the Limosilactobacillus reuteri H11 strain after 3h exposure to pH 3 ( 106 ). However, Limosilactobacillus reuteri L-3 displayed a moderate survival rate above 60% at pH 3.0, as observed by Wang et al. ( 112 ), and Lactiplantibacillus plantarum strains reported viability of 58% – 79% after 3 h exposure to pH 3 ( 113 ). The disparity in Lactobacillus isolate resistance to low pH may be due to the diverse low pH tolerance traits of several lactobacilli species sourced from different habitats and geographical regions ( 114 ). Highly acid-tolerant strains, such as Lactiplantibacillus plantarum and Limosilactobacillus reuteri , are important for modulating the oral and gut microbiota, promoting immune responses and reducing inflammation ( 103 , 105 ). Additionally, the Limosilactobacillus reuteri Mcyc strain isolated from this study may be crucial for developing a probiotic supplement for broiler chicken growth and development, while the Lactiplantibacillus plantarum To3a strain could be a promising candidate for creating a probiotic plant-based product, such as tomato juice, since they are host-adapted ( 115 ) . Among the 18 isolates obtained from the different samples, seven isolates, namely, Latilactobacillus sakei (Mcg, chicken gut), Lactobacillus johnsonii (Chxc4 and Chcx2, chicken gut), Lactobacillus johnsonii (Mcya and Mcyb, chicken gut), Latilactobacillus curvatus (Chcg, and Chcb, chicken gut), exhibited a survival rate ranging from 7% – 20% at pH 3.0, were excluded from the probiotic assay due to their poor tolerance to low pH, as HCl severely reduces the viable bacteria cell population in an acidic media ( 116 ). Khushboo et al.( 24 ) highlighted that for an organism to be considered probiotic, it must be able to resist the low acidic conditions (pH 1.0–3.0) necessary for survival in the host GIT. For this reason, isolates must display 50% tolerance to pH 3.0 for them to be considered resistant to low pH ( 87 ). Some bacterial cells lack a resistance mechanism when exposed to a stressful environment, such as a pH of 3.0, as proton influx at low pH disrupts the pH homeostasis mechanisms and reduces their intracellular pH ( 117 ). Cizeikiene & Jagelaviciute ( 118 ) mentioned that H + -ATPase activity is reduced for non-acid-tolerant strains when exposed to acidic conditions, consequently affecting the maintenance of an intracellular pH ( 118 ). At pH 3.5, a 0% survival rate was detected for Lactobacillus johnsonii UBLJ01 (vagina of healthy women) after 24 h of incubation via the traditional plate count method ( 119 ). In another study, the Lactobacillus isolates EKU 1 and EMB5, which were sourced from Ethiopian cottage cheese, showed 5% and 41.48% survival rates, respectively, at pH 3 after a 3 h incubation period ( 120 ). These poor acid-resistant isolates may be unable to grow and colonize in the stomach when utilized as probiotics; hence, they have low probiotic potential ( 121 ). Bile tolerance The human bile concentrations range from 0.3% to 0.5% ( 36 , 109 ), hence, any candidate probiotic bacteria must be able to survive the above-mentioned bile concentration ( 69 ), considering that bile salts are synthesized in the liver and disrupt the cell membrane, thus resulting in DNA damage and oxidative stress ( 122 ). The isolates Lactiplantibacillus plantarum (To3b and To3d, tomato), Limosilactobacillus reuteri (Mcyc, chicken gut), Lactobacillus johnsonii (Chx3b, Mcxb, and Mcxc, chicken gut), Lactiplantibacillus plantarum (Cu3e, cucumber) and Latilactobacillus curvatus (Pb2, banana) exhibited bile salt tolerances that were not significantly (p > 0.05) different between 0.3% bile salt and 0.5% bile salt. This finding indicates that the isolates possess good resistance to 0.3% and 0.5% bile salt, as they retain their cell viability ( 13 ). The ability of these lactobacilli isolates to resist the effects of bile salt toxicity may be due to bile salt hydrolases (BSHs), bile salt efflux, and modifications in the bacterial membrane composition ( 88 ). BSH sticks to bile salts and decreases their toxicity ( 122 ). With regards to Lactiplantibacillus plantarum , bile salt tolerance can be regulated by inducing BSH, modification of cell membrane composition and fluidity, preventing oxidative damage, and sustaining the proton motive force ( 123 ). Oh & Jung( 29 ) mentioned that several bile tolerance levels of three Lactiplantibacillus plantarum strains were linked to six proteins (GshR1, GshR4, Cfa2, Bsh1, OpuA, and AtpH) that may be crucial to the bile salt response and adaptation in this Lactobacillus species. Additionally, the bsh gene expression is upregulated in Lactiplantibacillus plantarum following bile exposure ( 124 ). The presence of polysaccharides on their outer cell membrane ( 87 ), S-layer protein protection and extracellular peptidoglycan, lipoteichoic acids, teichoic acid and proteins in Lactobacillus support the maintenance of the organism's cellular integrity ( 50 ). In accordance with our study, no significant differences in the tolerance rates of Lactiplantibacillus plantarum CCFM737 and RS4 were recorded when the bile salt concentration increased from 0% to 10% within a 12 h incubation time ( 88 ). Similarly, there were no significant differences observed in the growth rates of Lactiplantibacillus plantarum strains SWO3 and SW06, after 24 h of incubation with 0.3% and 1% bile salt ( 29 ). Zhang et al. ( 103 ) reported that no significant change in the bacterial viability rate was noted for Lactobacillus johnsonii incubated in 3% bile salts. Lactiplantibacillus plantarum strains Lp 21,30,31,32,33,35,41,52, and 55 displayed no significant changes when exposed to 0.3% and 0.5% bile salt after 24 h ( 94 ). However, none of the Lactobacillus plantarum strains sourced from tropical-grown fruits and vegetables were resistant to 0.2% and 0.3% bile salt after 24 h of incubation ( 77 ). In addition, Lactiplantibacillus plantarum and Limosilactobacillus reuteri strains sourced from the native chicken gut could not survive after 2 h in the presence of 0.3% (w/v) bile salt ( 51 ). The resistance of our study isolates to bile acid shows that they have good potential to survive bile salt stress conditions in the small intestine and remain viable to exert their probiotic functions ( 13 ). BSH activity BSH is an intracellular enzyme that catalyzes the hydrolysis of amide bonds between glycine or taurine and the steroid nucleus of bile salts ( 125 ). The ability of LAB to deconjugate bile salt is considered an indication of host-microbe interactions in the gut, which enable functional regulation of cholesterol metabolism ( 121 ). Three isolates, Lactiplantibacillus plantarum (To3a, tomato), Limosilactobacillus reuteri (Chcx3a, chicken gut), and Lactiplantibacillus plantarum (Cu3e, cucumber), exhibited the highest BSH activity with precipitation zones of 10–15 mm (++), and higher than the 1–10mm (+) of Lacticaseibacillus rhamnosus GG and other isolates. BSH is commonly distributed within the human and animal GIT and mainly originates from gram-positive intestinal bacteria such as Lactobacillus species ( 126 ). Lactobacilli are capable of performing various BSH enzymatic functions in the presence of bile salts, in particular, two BSH genes ( bsh 1 and bsh 3) out of 4 ( bsh 2 and bsh 4) in Lactiplantibacillus plantarum are linked with BSH activity ( 127 ). BSH enzymes play a vital role in reducing these toxic effects of glycoconjugated bile salts by allowing bile to enter the intestinal duodenum or microenvironments, when the pH decreases as a result of LAB ( 128 ). Furthermore, lactobacilli strains derived from the GIT, where bile is richly present, are more likely to be BSH positive ( 129 ). The overexpression and deletion of one or more bsh genes may be responsible for BSH activity in lactobacilli ( 130 ). The higher expression of the bsh 3 gene in Lactiplantibacillus plantarum is strongly associated with bile salt resistance ( 127 ). Our study corroborates a study that reported a large precipitation of up to 20 mm for Lactiplantibacillus plantarum F2 and 2F8 sourced from fresh figs and approximately 15 mm for Lactiplantibacillus plantarum FB13 derived from pricky pears against MRS agar supplemented with porcine bile ( 40 ). Another study observed that Lactiplantibacillus plantarum RC (raw cheese) and Lactiplantibacillus plantarum GV (guava) both presented precipitation zones > 1.5 cm greater than those of Lacticaseibacillus rhamnosus GG (1.0–1.5 cm) ( 131 ). Additionally, Limosilactobacillus reuteri TF-7, derived from pickled olives, exhibited a strong precipitation zone ≥ 13.1 mm against the sodium salt of taurodeoxycholic ( 99 ). Meanwhile, intestinal Limosilactobacillus reuteri LR20 strain displayed an intense level of BSH activity, as shown by dense precipitation of sodium taurocholate and sodium tauroglycocholate ( 96 ). Unlike our findings, Lactiplantibacillus plantarum V3F and Lacticaseibacillus rhamnosus GG both showed precipitation zones of up to 1.0–1.5 cm in a study by Boricha et al. (2019). Lactiplantibacillus plantarum 299v and DGIA1 showed low precipitation zones of 0 and 2 mm, respectively ( 132 ). In a different study, none of the Lactiplantibacillus plantarum strains isolated from fermented cabbage or cucumber ( 27 ), or rotten fruits or vegetables ( 14 ) showed any BSH activity. Some lactobacilli strains have different BSH-encoding genes; for example, Lactiplantibacillus plantarum possesses four genes ( bsh 1, bsh 2, bsh 3, and bsh 4) compared with others ( Limosilactobacillus reuteri , Latilactobacillus curvatus and Limosilactobacillus fermentum ), which contain only one or two bsh genes ( 125 , 130 ). The origin of the Lactobacillus strains plays an important role in BSH activity, as GIT or faecal-sourced microbes may have high BSH activity compared with bacteria sourced from plants or fermented foods ( 129 , 133 ). According to Kumar et al. ( 133 ), BSH enzymes in Lactobacillus isolates have different substrate specificities, considering that the majority of Bsh -active strains exhibit a preference for hydrolyzing glycocholate bile salt compared with taurocholate and taurodeoxycholate bile salts. Isolates with positive BSH activity have good potential for use as a dietary supplement ( 134 ). They also efficiently colonize the GIT and could be useful in reducing cholesterol levels in the body when administered as probiotics ( 35 ). However, the Lactobacillus johnsonii isolate (Mcxc, chicken gut) showed no precipitation zone in this study (Table 5 ) . The reason may be due to the absence of the bsh gene or its poor expression ( 130 ). Moreover, not all lactobacilli strains sourced from the GIT have BSH activity ( 129 ). Conversely, BSH activity was observed for Lactobacillus johnsonii 334 ( 135 ), Lactobacillus johnsonii NCD01693 ( 133 ), Lactobacillus johnsonii BFE 6128 and BFE 6154 ( 136 ) in a previous study. Lactobacillus isolates with no BSH activity are unable to survive and colonize the mucosal surfaces of the small intestines and hence cannot carry out cholesterol-lowering functions ( 137 ). Cholesterol-lowering ability of Lactobacillus isolates The cholesterol-lowering ability of Lactobacillus isolates is essential because it can help reduce harmful LDL cholesterol levels in the serum, thereby reducing the risk of cardiovascular disease ( 35 , 138 ). The isolates Lactiplantibacillus plantarum (To3a and To3d, tomato), Limosilactobacillus reuteri (Chx3a, Chicken gut), and Lactiplantibacillus plantarum (Cu3e, cucumber) exhibited significantly higher cholesterol-lowering abilities (p ≤ 0.05) compared with the Lacticaseibacillus rhamnosus GG, which showed a 41.81% reduction. Among these, Lactiplantibacillus plantarum (To3d, tomato) and Limosilactobacillus reuteri (Chx3a, chicken gut) demonstrated cholesterol-lowering capacities exceeding 50%, highlighting their strong potential as probiotic candidates for managing hypercholesterolemia ( 128 ). According to our findings, there was no relationship between cholesterol-lowering ability and the source of the strains investigated. Probiotic Lactobacillus strains can generate BSH enzymes that deconjugate bile salts, resulting in reduced cholesterol reabsorption and lower solubility ( 128 ). Ertürkmen et al. (2023) reported that these strains directly assimilate cholesterol into their cells and incorporate cholesterol into their cell membrane. This is later excreted from the body through faeces, hence lowering total cholesterol levels ( 139 ). The assimilation of cholesterol by lactobacilli isolates lowers the quantity of intestinal cholesterol that may be absorbed by enterocytes, hence reducing cholesterol absorption as well as its overall level ( 35 ). Cholesterol binds to the lactobacilli cell surface via adsorption ( 140 ), leading to a smaller amount of readily absorbed cholesterol in the intestine ( 141 ). Choi & Chang ( 142 ) attributed the unique chemical and structural properties of Lactobacillus plantarum cell wall compared with those of other LAB cell types as a possible reason for its strong cholesterol attachment. Short-chain fatty acids, which are mainly generated by gut microbes as a result of the fermentation of dietary fibre and resistant starch, can promote the conversion of cholesterol to bile acid, which, in turn, leads to a delayed atherosclerosis process ( 35 ). Moreover, EPS produced by certain LAB may bind bile acids and facilitate their excretion, which promotes the synthesis of bile acids from cholesterol, hence reducing the total circulating cholesterol ( 139 ). In line with our findings, Lactiplantibacillus plantarum strain CGMCC1.557 derived from vegetables exhibited a higher cholesterol assimilation ability (58%) than the 39% expressed by Lacticaseibacillus rhamnosus GG ( 143 ). Furthermore, Limosilactobacillus reuteri strain NCIMB 701089 assimilated more than 67% of cholesterol in their investigation, compared with the control (no probiotic), which accounted for 0%, and Lacticaseibacillus rhamnosus GG, which accounted for less than 50% ( 144 ). A higher cholesterol removal ability was observed in a previous study, where Lactiplantibacillus plantarum NCA4 (prickly pear) and 2F8 (fresh figs) exhibited cholesterol removal abilities of 66.04% and 81.24%, respectively, compared with the control after 24 h of incubation ( 40 ). Compared with those of the control, cholesterol assimilation by Lactiplantibacillus plantarum strains MYSDV5 and MYSDV2 sourced from traditional fermented food was reported to be 75.7% and 65.2% respectively ( 13 ). The Lactiplantibacillus plantarum isolate DLBSK207 (goat colostrum) displayed a cholesterol removal ability of 84.67%, followed by Lactiplantibacillus plantarum isolate DLBSH122 (mango fruit), with 65.34% cholesterol removal ability ( 141 ). In contrast, Lactiplantibacillus plantarum strains DUR 2, DUR5, and DUR8 sourced from Tempoyak presented a lower cholesterol removal ability (25.99%, 60.89% and 30.37%, respectively) than did Lacticaseibacillus rhamnosus GG (61.41%) ( 107 ). Moreover, Lactiplantibacillus plantarum NR74, which was isolated from kimchi, and Lacticaseibacillus rhamnosus GG showed similar cholesterol reduction rates of 47.8% and 48%, respectively, in a previous study ( 145 ). The importance of Lactiplantibacillus plantarum To3d and Limosilactobacillus reuteri Chx3a isolates, which exhibited a cholesterol-lowering ability of more than 50% suggests that they could be promising probiotic candidates for managing hypercholesterolemia because of their potent cholesterol-lowering effects ( 146 ), and their ability to minimize the risk of cardiovascular diseases ( 138 ). The EPS production ability The EPS production capability of some probiotic microorganisms is an attractive functional property, as it helps bacterial cells resist unfavourable environmental stress and allows effective gut colonization of mucosal surfaces ( 35 ), by increasing their auto-aggregation capacity ( 37 ). Most isolates, Lactiplantibacillus plantarum (To3a, To3b, and To3d, tomato), Limosilactobacillus reuteri (Chx3a, chicken gut), Latilactobacillus curvatus (Cu2f, cucumber), Lactiplantibacillus plantarum (Cu3e, cucumber) and Lactobacillus johnsonii (Mcxb, chicken gut), exhibited the ability to produce EPS. EPS are polymers made up of long chains of sugar molecules that have high molecular weights and differ in their properties and structures (147). They are secondary metabolites secreted into the extracellular environment during the growth and metabolism of microbes ( 148 , 149 ), and EPS molecules attach to the surface of the bacterial cell wall in the form of slime or a capsule ( 150 ). Lactobacillus species, including Limosilactobacillus reuteri , Lactiplantibacillus plantarum , Lacticaseibacillus rhamnosus and Latilactobacillus curvatus , are among the best EPS producers ( 151 , 152 ). In particular, species including Lacticaseibacillus rhamnosus and Lactiplantibacillus plantarum produce heteropolysaccharides (Heps) ( 153 ), which are synthesized within the cell and then carried out of the cell, whereas species such as Limosilactobacillus reuteri , Lactobacillus johnsonii and Latilactobacillus curvatus produce homopolysaccharides that are synthesized outside the cell, where a particular enzyme collects and assembles the sugar residues ( 154 ). Some proteins and enzymes participate in EPS production, which can be controlled by the regulation of gene expression of these proteins and enzymes ( 155 ). Pourjafar et al. ( 68 ) highlighted that the presence of the glycosyltransferase enzymes plays a significant role in the biosynthesis of EPS via available sugar units. Furthermore, certain gene cluster, such as eps , in Lactiplantibacillus plantarum , play key roles in EPS biosynthesis ( 148 ). Our results agree with the EPS production results observed for Limosilactobacillus reuteri isolates (SHA101, SHA103, SHA104, SHA107, SHA111 and SHA113) and Lactobacillus johnsonii (SHA105) ( 18 ), Latilactobacillus curvatus SJTUF 62116 ( 150 ), Latilactobacillus curvatus SHA2-3B ( 156 ), and Lactobacillus johnsonii PUMSKGRI ( 36 ). Similarly, Lactiplantibacillus plantarum 53 ( 104 ), Lactiplantibacillus plantarum KMUDR7 ( 121 ) and Lactiplantibacillus plantarum 47FE ( 157 ) have been reported to show EPS production capacity. The wide range of sources of EPSs results in variation in their structural composition and physicochemical properties, and these dissimilarities confer different biological functions( 149 ) Moreover, the differences in EPS may vary owing to monosaccharide conformation, the connections between builder units, charge, the existence of frequent side-chains, and replacement ( 68 ). In addition to the factors mentioned above, the conditions of the environment, such as pH, temperature, nutrient composition and the carbon source utilized by the bacterium ( 92 , 158 ), the transcriptional gene levels necessary for the production of EPS( 159 ) and the over-expression of the esp gene clusters in the bacterium may significantly influence EPS production capacity. EPS produced by these strains may be leveraged by the fermented food industry as bio-thickeners, due to their water-binding ability, thereby improving the rheological features, and further playing an important role in food processes such as viscosity, emulsification, and flocculation ( 160 ). In the dairy industry, Lactobacillus EPS may play a primary role when used as thickeners and texturizers to enhance food product viscosity, as they may also act as stabilizers, reduce syneresis, and interact with different milk constituents ( 155 ). Additionally, EPS producing Lactobacillus presents potential as a replacement for chemical additives ( 161 ). Conversely, the Lactobacillus isolates Lactobacillus johnsonii (Chx3b, chicken gut), Limosilactobacillus reuteri (Mcyc, chicken gut), Latilactobacillus curvatus (Pb2, banana), and Lactobacillus johnsonii (Mcxc, chicken gut) were unable to produce EPS, suggesting potential variability in EPS production among the strains studied. The absence or deletion of the crucial cluster gene ( esp ), essential for EPS biosynthesis, may result in low EPS yields ( 162 , 163 ). A mutant strain or alterations in the eps cluster genes could significantly impair the production level of EPS in the Lactobacillus strain ( 162 , 164 ). Moreover, the absence of the glycosyltransferase enzyme necessary for EPS biosynthesis may result in a lack of EPS production ( 68 ). Other factors, such as nutrient composition, especially starvation or an oversupply of essential nutrients such as nitrogen, sugars, carbon dioxide, etc., could alter EPS biosynthesis ( 92 ). For example, Fuso et al. ( 154 ) mentioned that a low EPS production yield in Lactobacillus depends on the kind of sugar (glucose, maltose, lactose, galactose, mannose, ribose, glucosamine and fructose) used as a carbon source. According to Mıdık et al. ( 165 ), LAB EPS production is negatively affected by an increase in temperature from 30–37°C, extended incubation time above 48 h, pH 5.0 and 7.0 and increased NaCl concentration of 3% and 6%. In contrast to our findings, Lactobacillus johnsonii PUMSKGRI ( 36 ) and Lactobacillus reuteri DUR12 ( 107 ) demonstrated EPS production ability. Lactobacillus isolates with a negative EPS production capacity may play an insignificant role when used in the food industry as viscous, stabilizing and emulsifying agents ( 92 ). DPPH scavenging activity of Lactobacillus strains An imbalance between high levels of reactive oxygen species (ROS) and low antioxidant capacity is known as oxidative stress ( 166 , 167 ). ROS are generated during normal metabolic processes and play essential roles in various biological processes ( 168 ). However, excessive oxidative stress has been linked to several chronic conditions such as cancer, diabetes, hypertension, ulcers, aging, and atherosclerosis ( 38 ). Interestingly, some probiotic microbes produce bioactive compounds that can counteract oxidative stress via the prevention of ROS formation through specific molecular mechanisms ( 169 ). Therefore, identifying these organisms capable of exerting antioxidant activity in the human body is desirable. The DPPH scavenging activity of Lactiplantibacillus plantarum (To3d, tomato) (24.45%) was significantly (p ≤ 0.05) higher than that of Lacticaseibacillus rhamnosus GG (23.20%), whereas the activity of the isolate Limosilactobacillus reuteri (Mcyc, chicken gut) was not significantly different (P > 0.05). The Lactobacillus genus can generate antioxidant enzymes (catalase, superoxide dismutase (SOD), and nicotinamide adenine dinucleotide phosphate (NADH) oxidase and NADH peroxidase), or non-enzymatic antioxidants (Mn 2+ , vitamins C and E, tocoferols, glutathione), which depend on small molecule activity together with enzymes to neutralize excess ROS ( 101 , 170 ). For example, catalase plays a valuable role in alleviating oxidative stress by degrading hydrogen peroxide (H 2 O 2 ) ( 171 ). Lactobacilli can produce EPS and organic acids (lactic acid, acetic acid, etc.), which contribute to alleviating the effects of ROS and enhancing microbial antioxidant activity ( 172 ). This is due to its ability to donate electrons or hydrogen atoms, eliminate free radicals, and chelate metal ions ( 161 ). In addition, the antioxidant molecules (C30 carotenoid exopolysaccharides, ferulic acid and lactate) produced by these Lactobacillus may also mitigate free radical formation through a neutralization reaction ( 173 ). Certain Lactobacillus species possess oxidative stress resistance genes and proteins essential for redox mechanisms, such as thioredoxin antioxidant proteins expressed by certain Lactiplantibacillus plantarum and Lacticaseibacillus casei strains or the catalase gene ( kat A) expressed by Latilactobacillus sakei ( 170 ). Additionally, they can efficiently prevent high levels of ROS formation and reduce ROS stress by chelating with metal ions (Fe2+) ( 172 , 174 ). Feng & Wang ( 174 ) highlighted that these microbes possess a ROS-binding (glutathione and thioredoxin) system that maintains intracellular dithiol/disulfide homeostasis in bacterial cells, playing a vital role in protection against oxidative stress and thereby protecting bacteria from oxidative damage. The antioxidant capacity of Lactobacillus isolates corresponds with findings from earlier research, where Lactiplantibacillus plantarum strain DUR8 exhibited DPPH scavenging activity (73.36%), which was higher than that of the reference strain Lacticaseibacillus rhamnosus GG (59.15%) ( 107 ). Another study revealed that Lactiplantibacillus plantarum 27156, 27195, 27197 and 27319 displayed a maximum DPPH scavenging rates of 34.85%, 34.85%, 30.3% and 39.3%, respectively, compared with the 28.24% reported for Lacticaseibacillus rhamnosus GG ( 175 ). DPPH scavenging activity was 30.51% for Lactiplantibacillus plantarum 200655, whereas it was 27.93 % for Lacicaseibacillus rhamnosus GG ( 166 ). Conversely, Kim et al. (2020) reported that the DPPH scavenging rate of Limosilactobacillus reuteri MG505 was significantly higher (33.5%) than that of Lacticaseibacillus rhamnosus MG316 (22.2%). Probiotic Lactobacillus organisms that have high antioxidant activity may serve as antioxidant supplements that can reduce ROS levels in the host’s body tissues and considerably reduce the risk of oxidative damage in humans and animals when ingested ( 101 ). Additionally, treatment with probiotic Lactobacillus antioxidant supplements may prevent or manage ailments, such as inflammatory bowel disease, by downregulating the expression of inflammatory factors while promoting an increase in related antioxidant enzymes, possibly through the activation of various host signalling pathways ( 170 ). The DPPH scavenging activities of the Lactobacillus isolates, Lactiplantibacillus plantarum (To3b and To3b, tomato), Limosilactobacillus reuteri (Chx3a, chicken gut), Lactobacillus johnsonii (Chx3b, chicken gut), Latilactobacillus curvatus (Cu2f, cucumber), Lactiplantibacillus plantarum (Cu3e, cucumber), and Latilactobacillus curvatus (Pb2, banana), were significantly (p ≤ 0.05) lower than those of the reference strain Lacticaseibacillus rhamnosus GG (23.20%). The antioxidant activity of Lactobacillus species differs as a result of dissimilarities in metabolic pathways, such as the recombination DNA repair pathway, microbial metabolism, pyruvate metabolism, etc. ( 171 ). According to Zhao et al. ( 170 ), only a few species possess oxidative stress resistance genes and proteins important for redox mechanisms. In some cases, some Lactobacillus strains may not possess SOD genes, suggesting that different species encode different redox–related genes and have different redox systems ( 174 ). Furthermore, certain lactobacilli demonstrate low expression or loss of the activities of catalase and the CAT antioxidant enzyme, especially under stress conditions like pH and elevated temperatures ( 173 ). Prete et al. ( 176 ) highlighted that some microbial strains that produce low EPS may possess reduced antioxidant activity compared with high EPS producers. Our results are consistent with the findings of Kim et al. ( 38 ), who reported that Limosilactobacillus reuteri MG505 had a DPPH radical scavenging rate of 33.5%, whereas Lacticaseibacillus rhamnosus MG316, had a DPPH rate of 22.2%. Compared with the control, ascorbic acid (37.15%), Latilactobacillus curvatus MG5020 showed a lower DPPH activity of 5.32% ( 167 ). Vougiouklaki et al. ( 177 ) observed that the DPPH scavenging rate of Lactiplantibacillus plantarum ATCC 14917 (77.53%) was lower than that of Lacticaseibacillus rhamnosus GG (83.41%) after 210 min. Similarly, the Lactiplantibacillus plantarum isolates LpJ2, LpJ5, LpJ6, LpJ7, LpJ8, LpJ18, and LpJ20 presented low DPPH activity that was < 50% after 30 min, as compared with Lacticaseibacillus rhamnosus GG (51.52%) ( 109 ). Differences in the antioxidant potential of different Lactobacillus strains suggest that the radical-scavenging activity of the microbes examined may be due to the strain-specific nature of Lactobacillus ( 178 ). Debnath et al.( 109 ) emphasized that cell surface components, such as extracellular polysaccharides, contribute to variations in the DPPH scavenging activities of microbes. Furthermore, the difference in the scavenging rates of microbial cultures may be due to the metabolic activity of different bacterial species ( 168 ). Lactobacillus isolates with weak antioxidant activity may be less efficient at neutralizing reactive free radicals when used as probiotic antioxidant supplements ( 168 ). Antimicrobial activity of Lactobacillus isolates The probiotic Lactobacillus , which has antimicrobial activities, supports gut health by suppressing pathogens that cause diseases in the host and offers natural alternatives to antibiotics ( 16 ). The isolates, Lactiplantibacillus plantarum (To3a, To3b, and To3d, tomato), exhibited antimicrobial activity similar to that of the reference strain Lacticaseibacillus rhamnosus GG, with an inhibition zone of 10 to 15 mm (++) or more, against E. coli , S. aureus , B. cereus and L. monocytogenes . The high antimicrobial activity of these Lactobacillus strains may be attributed to the production of metabolites such as bacteriocins, H 2 O 2 , and organic acids (lactic and acetic acid) that can inhibit these pathogens ( 45 , 90 ). For example, bacteriocins prevent pathogenic cells by disrupting their cell membrane and/or forming pores that lead to the death of the cells via a fast-acting mode of action ( 179 , 180 ). Notably, Lactiplantibacillus plantarum produces organic acids that decrease the pH of the environment, consequently inhibiting the growth of pathogenic bacteria ( 116 ). Additionally, they can prevent the growth of pathogenic organisms via competitive effects, enhancing the permeability of the thin outer membrane, changing the intracellular osmotic pressure, and preventing the synthesis of macromolecules ( 181 ). High EPS-producing lactobacilli strains may promote their antimicrobial activity as a result of metal chelation, inhibition of nutrient uptake via barrier formation, cell wall or cytoplasmic membrane disturbance, suppression of the synthesis of mRNA and proteins, impairing cell division or decomposing DNA ( 114 ). Furthermore, lactobacilli with high auto-aggregative potential can inhibit pathogenic organisms from colonizing the GIT via the formation of a barrier through auto-aggregation ( 182 ). Besides that, S-layer protein A (SlpA) in Lactobacillus acidophilus , for instance, hinders bacterial infection by blocking the activity of the cellular receptor DC-SIGN and murein hydrolase ( 183 ). Our findings align with those of a previous study, where the diameters of the clear zones of Lactiplantibacillus plantarum KU200656 were reported to be 31.44 mm and 32.71 mm, against E. coli and L. monocytogenes , respectively, which are slightly similar to those of Lacticaseibacillus rhamnosus GG, with inhibition zones of 29.00 mm and 23.13 mm, respectively ( 184 ). A previous study observed that Lactiplantibacillus plantarum S2-5 and Lacticaseibacillus rhamnosus GG demonstrated similar diameters of inhibition zones against E. coli (2–5 mm) and S. aureus (< 2 mm) ( 185 ). Additionally, Lactiplantibacillus plantarum KA18 and KLAB5 displayed the same inhibition zone as Lacticaseibacillus rhamnosus GG at 15–20 mm against L. monocytogenes KACC 10764 and S. aureus KCCM 40510 and 11–14 mm against E. coli K99 KCTC 2617 ( 186 ). In another study, overnight cultures of Lactiplantibacillus plantarum CGMCC 1.557 and Lacticaseibacillus rhamnosus GG exhibited similar inhibition zone diameters (> 6 mm) against E. coli , B. cereus and S. aureus ( 143 ). In contrast, Lacticaseibacillus rhamnosus GG exhibited robust inhibition zones of 28 mm, 27 mm, 34 mm and 29 mm against L. monocytogenes , S. aureus , B. cereus and E. coli , respectively, whereas Lactiplantibacillus plantarum V3F presented inhibition zones of 17 mm, 25 mm, 17 mm and 40 mm against L. monocytogenes , S. aureus , B. cereus , and E. coli, respectively ( 15 ). The differences in the antimicrobial effects observed in this study could be attributed to acidic products such as lactic and acetic acids, or bacteriocins, among others ( 187 ). Furthermore, the disparity in results may be due to the varying sensitivities of target strains to antimicrobial compounds produced by Lactobacillus species and the detection methods used ( 188 ). Oral administration of probiotic Lactobacillus strains with strong antagonistic effects may be used to inhibit the growth and colonization of harmful pathogenic microbes in the host GIT and fermented products ( 90 ). Moreover, these lactobacilli or their antimicrobial compounds are great tools that may be applied in the control of food spoilage and/or pathogenic organisms, to improve food safety and extend bio-preservation ( 189 ). The other Lactobacillus isolates- Limosilactobacillus reuteri (Chx3a, chicken gut), Lactobacillus johnsonii (Chx3b, chicken gut), Latilactobacillus curvatus (Cu2f, cucumber), Lactiplantibacillus plantarum (Cu3e, cucumber), Latilactobacillus curvatus (Pb2, banana), Limosilactobacillus reuteri (Mcyc, chicken gut), and Lactobacillus johnsonii (Mcxb, and Mcxc, chicken gut) exhibited lower antimicrobial activity, with inhibition zones below the 10–15 mm (++) range against one or more of the tested pathogens. Some lactobacilli produce few antibacterial compounds, such as H 2 0 2 and bacteriocin, thus resulting in weak inhibitory activity against pathogens ( 190 ). The presence or lack of specific bacteriocin genes among Lactobacillus strains influences their ability to inhibit a wide range of pathogens ( 188 ). Moreover, the antibacterial effects of any Lactobacillus strains are notably reduced when the culture medium or the environmental conditions are at neutral or high pH levels ( 188 , 191 ). Similarly, Latilactobacillus curvatus KG 12 − 1 exhibited no inhibition zones (0 mm) against E. coli , B. cereus or S. aureus in their research study ( 192 ). Lactobacillus johnsonii and Limosilactobacillus reuteri expressed 9.0 mm and 9.1 mm inhibition zones, respectively, against E. coli D7 ( 54 ). Moreover, the diameter of the inhibition zones for Lactiplantibacillus plantarum S2-5 and S4-1 was noted to be between 2–5 mm and 6–8 mm, respectively, against E. coli O157, while that of S. aureus AC1 was observed to be 2 mm and 0 mm ( 185 ). Similarly, Lactobacillus johnsonii (SHA 105) exhibited a 9.3 mm inhibition zone, while Limosilactobacillus reuteri (SHA 101, 103, 104, 107, 111 and 113) had antimicrobial activity ranging from 11.1 mm to 14.2 mm against E. coli ATCC 25922 ( 18 ). Conversely, some researchers revealed that Latilactobacillus curvatus P99 exhibited higher antimicrobial activity with an inhibition zone of 23 mm against E. coli ATCC 8739, 16.5 mm against B. cereus ATCC 11778 and L. monocytogenes ATCC 7644 and 15 mm against S. aureus ATCC 25923 ( 193 ). Limosilactobacillus reuteri H11 exhibited a larger inhibition zone against E. coli ATCC 25922 (21.33 mm), S. aureus ATCC 6538 (19.33 mm) and L. monocytogenes ATCC 10403 (> 15 mm) ( 106 ). The degree of pathogenic inhibition by Lactobacillus isolates varies due to the strain-specific nature of Lactobacillus ( 194 ). Lactobacillus isolates with low antibacterial activity may possess poor bacteriostatic potential and are unattractive for probiotic application in terms of preventing or reducing the growth and colonization of opportunistic intestinal microbes ( 195 ). Cell auto-aggregation The ability of the same bacterial strains to bind to themselves has been linked to enhanced probiotic survival, colonization, and pathogen exclusion, making it a desirable trait for selecting potential probiotic strains ( 87 ). Moreover, auto-aggregation is a probiotic feature that involves bacterial entrapment in an aggregated form, which allows for stability in the GIT, resulting in less exposure to inhospitable intestinal conditions ( 39 ). The isolates Lactiplantibacillus plantarum (To3a, tomato), Limosilactobacillus reuteri (Chx3a, chicken gut), Lactobacillus johnsonii (Chx3b, chicken gut), Latilactobacillus curvatus (Cu2f, cucumber), and Limosilactobacillus reuteri (Mcyc, chicken gut) exhibited a significant (p ≤ 0.05) increase in auto-aggregation (%) after 2 and 4 h incubation compared with the Lacticaseibacillus rhamnosus GG. Additionally, the auto-aggregation (%) of these isolates was significantly (p ≤ 0.05) greater than that of the reference strain after 2 and 3 h of incubation. The presence and/or high expression of cell surface proteins in Lactobacillus , such as the sortase-dependent protein from Lactiplantibacillus plantarum CMPG5300( 196 ) or the serine/threonine-rich protein from Lactiplantibacillus plantarum NCIMB 8826, are contributory factors that promote high cell auto-aggregation ( 197 ). Auto-aggregation is associated with cell surface hydrophobicity; hence, high surface hydrophobicity in lactobacilli may facilitate their ability to self-clump or aggregate readily ( 198 ). Additionally, Lactobacillus auto-aggregation activity is due to the specific strain and not the species; as a result, Lactobacillus strains exhibit a wide range of auto-aggregation percentages, as seen in Lactiplantibacillus plantarum from 29.32% to 80% ( 199 ). An earlier report mentioned that a reduction in EPS production in Lactobacillus johnsonii resulted in a high increase in auto-aggregation and HT29 cell adhesion ( 200 ). Lactobacillus aggregation ability may be influenced by many factors, such as the culture conditions of the bacteria, environmental conditions (time, temperature, pH value) and bacterial pre-treatment before aggregation ( 199 ). Auto-aggregation is dependent on environmental conditions and incubation time, as the degree of aggregation increases with increasing incubation time ( 198 ). According to Zawistowska-Rojek et al. ( 201 ), a higher aggregation rate is observed in longer bacterial cells due to their larger surface area than those with shorter cells or spherical shapes. Our observations agree with the results of a previous study in which three strains of Lactiplantibacillus plantarum LpF, LpG and LpH showed high auto-aggregation abilities of 15.8%, 20.5% and 15.4%, respectively, compared with the 13.1% reported for Lacticaseibacillus rhamnosus GG after a 2 h incubation period ( 201 ). Greater cell auto-aggregation was reported for Limosilactobacillus reuteri DUR12 at 47.76%, Lactiplantibacillus plantarum DUR2, DUR5 and DUR8 at 52.24%, 48.10%, and 49.88%, respectively, than for Lacticaseibacillus rhamnosus GG (40.55%) after 5 h ( 107 ). Furthermore, Latilactobacillus curvatus JBCC38, JBCC93, and JBCC99 exhibited auto-aggregation abilities of 37.8%, 36%, and 31.2%, respectively, which was greater than the 20.1% reported for Lacticaseibacillus rhamnosus GG after 2 h ( 64 ). In their study, Lactobacillus rhamnosus GG auto-aggregation values were recorded as 86%, whereas Lactobacillus plantarum strain OR had 92% auto-aggregation percentage after 24 h ( 131 ). On the contrary, Lactobacillus johnsonii PF01 presented a lower auto-aggregation ability (34%) after 24 h of incubation than did Lacticaseibacillus rhamnosus GG (36%) ( 202 ). A lower degree of auto-aggregation was reported for Limosilactobacillus reuteri strains, ranging from 6.3% to 24.0% after 3 h and 14.4% to 29.0% after 6 h ( 203 ). In a different study, a similar auto-aggregation value was observed for Lactiplantibacillus plantarum CGMCC 1.557 (34%) and Lacticaseibacillus rhamnosus GG (34%) after a 2 h ( 143 ). Based on this study result, Lactobacillus isolates with high auto-aggregation ability may have increased potential to adhere to and colonize the host's GIT when consumed as probiotics ( 204 ). Additionally, the high auto-aggregation value among Lactobacillus species highlights the high potential of these bacteria to inhibit pathogens from colonizing the host GIT ( 161 , 205 ). Considering that auto-aggregation is pivotal for biofilm development, these bacteria could be exceptional probiotic candidates when incorporated into functional foods ( 194 ). Cell surface hydrophobicity Cell surface hydrophobicity determines the ability of probiotic Lactobacillus cells to interact with and adhere to surfaces such as epithelial cells, a key feature of probiotics ( 194 , 206 ). The isolates Latilactobacillus curvatus (Cu2f, cucumber), Lactiplantibacillus plantarum (Cu3e, cucumber), and Limosilactobacillus reuteri (Mcyc, chicken gut) exhibited significantly higher cell surface hydrophobicity values (p ≤ 0.05) than did Lacticaseibacillus rhamnosus GG (53.33%). In contrast, the isolate Lactobacillus johnsonii (Chx3b, chicken gut) displayed cell surface hydrophobicity (%) not significantly different (p > 0.05) from that of Lacticaseibacillus rhamnosus GG. The higher degree of cell surface hydrophobicity observed in the Lactobacillus isolates Latilactobacillus curvatus (Cu2f, cucumber), Lactiplantibacillus plantarum (Cu3e, cucumber), and Limosilactobacillus reuteri (Mcyc, chicken gut) can be attributed to the presence of cell surface proteins in lactobacilli, including S-layer proteins, which facilitate hydrophobic interactions with the host cell surface ( 207 ). For example, the cell surface mucus- binding protein ( MapA ) found in Limosilactobacillus reuteri ( 208 ), and the surface protein found in Lactiplantibacillus plantarum HC-2 ( 209 ). In addition, an increase in the rate of cell surface hydrophobicity has been linked to certain lactobacilli producing highly hydrophobic EPS, which results in increased biofilm formation ( 210 ). Previous researchers have noted a similar trend of broad variations in hydrophobicity values among the Lactobacillus strains studied ( 36 , 41 , 95 , 104 , 211 , 212 ). In agreement with our findings, 15 Lactiplantibacillus plantarum isolates (LpJ1-7, LpJ10-13, and LpJ16-20) displayed higher xylene surface hydrophobicity (19.58% to 62.59%) than did Lacticaseibacillus rhamnosus GG (15.96%) after 30 min of incubation ( 109 ). The n-hexadecane hydrophobicity values observed for 9 Lactiplantibacillus plantarum isolates (Fb, RC, GV, OR, SG, CK, GP, HB, and OP) were reported to range from 2% to 27%, whereas Lacticaseibacillus rhamnosus GG exhibited 13% hydrophobicity in a previous study ( 131 ). Hydrophobicity values of 40% were recorded for Limosilactobacillus reuteri I2 from the poultry GIT ( 20 ), 61.15% for Limosilactobacillus reuteri L-3 sourced from the chicken gut ( 112 ), and 76% for Limosilactobacillus reuteri K18 (colostrum) ( 203 ). The wide differences in the cell surface hydrophobicity of the Lactobacillus isolates in this study may be due to variations in the level of expression of cell surface proteins among lactobacilli strains and environmental conditions, which could impact the expression of surface proteins ( 106 , 109 ). In addition, the bacterial cell growth phase may contribute to affecting the surface hydrophobicity of these bacteria ( 90 ). Other compounds (core oligosaccharides, lipoteichoic acid, lipids, surface fibrils and several fimbriae) contribute to differences in the cell surface hydrophobicity ( 198 ). The high cell surface hydrophobicity of some bacterial isolates in this study could play a pivotal role in promoting the adhesion and colonization of lactobacilli to the epithelium and mucosal surfaces of the host ( 36 , 106 ). The low cell surface hydrophobicity (%) of the Lactobacillus isolates, Lactiplantibacillus plantarum (To3a, To3b and To3d, tomato), Limosilactobacillus reuteri (Chx3a, chicken gut), Latilactobacillus curvatus (Pb2, banana), and Lactobacillus johnsonii (Mcxb and Mcxc, chicken gut) may be attributed to the absence of relevant structural and compositional features of their cell surfaces (S-proteins, adhesins, and polysaccharides), which reduce their ability to interact with hydrophobic substrates ( 213 ). The surface proteins of several lactobacilli, including Lactobacillus crispatus and Lactobacillus acidophilus , whose ability to bind to host epithelial cells decreases after removal or disruption of the S-layer proteins ( 214 ). The abundance and presence of the EPS layer at the bacterial cell surface, which completely covers the cell and shields surface-bound proteins, could lower the hydrophobicity of lactobacilli ( 200 ). The low percentage of cell surface hydrophobicity exhibited by lactobacilli strains in this study aligns with the observed xylene hydrophobicity value for 10 Limosilactobacillus reuteri strains (PL103, PL110, PL111, PL112, PL113, PL114, PL115, PL116, PL118, and PL120) and one Lactobacillus johnsonii strain (PL105) derived from the native chicken GIT, which was recorded to be < 20% ( 51 ). An earlier study revealed that the xylene cell surface hydrophobicity exhibited by Latilactobacillus curvatus BSF206 were less than 30% ( 215 ). A hydrophobic index of less than 40% was observed for Lactiplantibacillus plantarum B20, whereas more than 50% hydrophobic indices were recorded for Lacticaseibacillus rhamnosus GG ( 90 ). Lactobacillus isolate with a low percentage of cell surface hydrophobicity may not effectively colonize and persist in the host GIT when they are used as probiotics ( 216 ). Antibiotic susceptibility Owing to the possibility of the transfer of antibiotic resistance genes to intestinal pathogenic bacteria, serious safety concerns regarding probiotics sheltering antibiotic resistance genes still exist ( 41 , 87 ), because of the potential for drug resistance in pathogenic organisms ( 181 ). All the Lactobacillus isolates were susceptible to antibiotics: penicillin G (10 µg), chloramphenicol (30 µg), erythromycin (15 µg), and ampicillin (10 µg). The probable reason may be that most Lactobacillus species lack β-lactamase enzymes, which are sensitive to some of these antibiotics ( 217 ). The thick peptidoglycan layer in these bacterial cell walls is vulnerable to antibiotics, such as penicillin and ampicillin ( 218 ). Additionally, the absence of a resistance gene in some lactobacilli strains (such as the gene mec A, which is responsible for penicillin resistance; the chloramphenicol resistance gene cat ; or the erythromycin resistance gene erm B) contributes to vulnerability to these antibiotics ( 217 ). The tolerance of Lactobacillus species to these antibiotics is consistent with the findings of these previous studies ( 15 , 16 , 41 , 45 , 51 , 219 ). The high susceptibility of our Lactobacillus isolates to these antibiotics suggests they are safe for use as probiotics, since they may not contribute to the spread of antibiotic resistance genes in the host GIT microbiota ( 161 ). Conversely, all the Lactobacillus isolates were resistant to aminoglycoside antibiotics: kanamycin (30 µg), gentamicin (10 µg), and streptomycin (10 µg) (Table 10 ). This may be due to the presence of aminoglycoside resistance genes such as the aph(3′) - IIIa gene, which is responsible for kanamycin resistance; the ant ( 6 )- Ia ( aadE ) gene, which is involved in streptomycin resistance; and the ant(4′) - Ia gene, which is responsible for neomycin resistance in lactobacilli strains ( 220 ). As mentioned in an earlier study, the ability of the Lactobacillus genus to resist these antibiotics may also be attributed to the low impermeability of its cell surface to aminoglycoside antibiotics such as kanamycin ( 70 ). Moreover, the lack of cytochrome-mediated electron transport, which promotes drug absorption, is another reason why these bacteria may be intrinsically resistant to this group of antibiotics ( 70 , 87 ). This finding is in accordance with some studies ( 18 , 41 , 181 , 211 , 221 ) reporting that Lactobacillus strains are resistant to aminoglycoside antibiotics. Cell culture conditions, the position of the resistance gene, and the participation of other mechanisms might affect the degree of antibiotic resistance ( 182 ). Lactobacillus strains may survive in the gut even after treatment with aminoglycoside antibiotics and restore the intestinal microflora ( 222 ), and when they are co-administered with an antibiotic, they might reduce the gut side effects linked with the antibiotics ( 69 ). In this present study, 6 Lactobacillu s isolates, Limosilactobacillus reuteri (Chx3a, chicken gut), Lactobacillus johnsonii (Chx3b, chicken gut), Latilactobacillus curvatus (Pb2, banana), Limosilactobacillus reuteri (Mcyc, chicken gut), Lactobacillus johnsonii (Mcxb and Mcxc, chicken gut), were resistant to tetracycline (Table 10 ). The high resistance rate to tetracycline among chicken gut samples (Chcx3a, Chcx3b, Mcyc, Mcxb, and Mcxc) in this study may be due to the common use of tetracycline antibiotics by poultry farmers in the Johannesburg metropolitan city in South Africa. Motey et al. ( 223 ) highlighted that Lactobacillus species, which are resistant to tetracycline, may harbour transferable tet genes such as tet ( M ) and tet ( W ), which have been reported in other studies of lactobacilli. Tetracycline resistance among lactobacilli may occur via ribosomal protection proteins, efflux pumps and direct enzymatic inactivation ( 70 ). Some studies have revealed that all Lactobacillus isolates examined in their study were resistant to tetracycline ( 14 , 18 , 86 ). Furthermore, tetracycline resistance has been observed in studies of Lactobacillus isolates sourced from fermented cabbage and cucumber ( 27 ), rectal swabs of 21-day-old chickens, their faecal and feed samples ( 45 ). Our findings imply that the tetracycline-resistant Lactobacillus isolates may be resistant to these antibiotics when used for probiotics, as they encourage the spread of antibiotic resistance genes in the gut microbiota and the environment ( 217 ). Most of the isolates (66.7%) in this study were resistant to vancomycin (Table 10 ). The resistance of Lactobacillus strains to vancomycin is intrinsic and non-transferable due to the absence of D-Ala-D-Lac target modification ( 161 ). The presence of the vancomycin-resistance gene van X ( 217 , 218 ), which encodes D-Ala-D-Ala dipeptidase, which is crucial for cell wall synthesis, is also a contributing factor ( 224 ). The absence of a specific drug target or cell wall impermeability also contributes to their intrinsic resistance ( 224 ). This finding was closely consistent with a study by Yuksekdag et al. ( 17 ), which reported that eleven out of eighteen Lactobacillus strains isolated from the chicken GIT were resistant to vancomycin. Nonetheless, some researchers have reported a relatively high incidence of vancomycin resistance among Lactobacillus strains ( 16 , 18 , 20 , 27 , 211 , 221 ). Vancomycin resistance among Lactobacillus isolates may assist in the prevention of antibiotic-associated diarrhoea, and in some cases, it may restore gut microflora ( 224 ). Blood haemolysis Lactobacillus strains intended for probiotic use must not cause lysis of red blood cells; thus, the absence of haemolytic activity is a critical safety requirement for probiotic strains ( 225 ). As observed in this study, none of the isolates showed haemolysis on blood agar. This may be attributed to the absence of haemolysin-encoding genes, which are responsible for red blood lysis in bacterial cells ( 179 ). Some Lactobacillus strains may harbour a gene encoding protein that may imitate haemolysins ( 179 ), and the environmental conditions (oxidative stress, low pH, and temperature), as well as the growth medium used, may result in discrepancies in some study findings ( 87 ). Our results corroborate with many studies that have confirmed that Lactobacillus is non-haemolytic ( 13 , 15 , 30 , 104 , 226 ). All the isolates tested met the safety criteria for probiotic use; therefore, they are safe for human consumption as probiotics ( 161 ). DNase activity DNases are extracellular endonucleases that cleave the phosphodiester bond in the backbone of DNA, releasing free nucleotides and phosphates, thus disrupting cell functionality ( 227 ). None of the examined Lactobacillus isolates demonstrated DNase activity, indicating a lack of potential for nucleic acid degradation and associated pathogenicity ( 228 ), as shown in Table 11 . These results agree with studies that reported that Lactobacillus strains showed no DNase activity ( 13 , 37 , 226 ). Hence, this finding proves the safety of these Lactobacillus strains for human use as probiotics in the food industry, considering that the degradation of host DNA is often associated with virulence in pathogenic bacteria ( 228 ). Caco-2 cells adhesion capacity Potential probiotic Lactobacillus species must adhere to and colonize the mucosal surface and epithelial cells to ensure prolonged persistence in the GIT and exert beneficial health effects ( 117 , 204 ). Accordingly, Caco-2 cells are widely used in vitro to assess probiotic adhesion capacity due to their close resemblance to mature intestinal epithelial cells ( 86 ). Lactobacillus johnsonii (Chx3b, chicken gut) and Lactiplantibacillus plantarum (To3d, tomato) exhibited higher Caco-2 cells adhesion capacities (15.29% and 12.87%, respectively) than Lacticaseibacillus rhamnosus GG (11.40%), whereas isolate Limosilactobacillus reuteri (Mcyc, chicken gut) presented a slightly lower adhesion capacity of 10.53% (Table 12 ). The reason for this can be attributed to the structural differences and cell wall constituents (polysaccharides, fimbriae or pili, adhesins, surface proteins, and other components) of Lactobacillus species, which increase their ability to attach strongly to the host epithelial cells ( 203 , 216 ). As mentioned previously, the surface EF-Tu moonlighting protein of Lactiplantibacillus plantarum 423 ( 183 ) and the mucus-binding protein of Lacticaseibacillus rhamnosus GG promotes high cell adhesion ability ( 225 ). The high auto-aggregation ability of lactobacilli has also been linked to their high capacity to adhere to epithelial cells and prevent the colonization of pathogens in the GIT ( 161 , 200 ). Moreover, Lactobacillus strains with high surface hydrophobicity values have a greater capacity to adhere to epithelial cells ( 106 ). In addition, EPS or lipoteichoic acids produced on the cell walls of lactobacilli contain adhesive molecules, which can enhance their adhesion capacity to the intestinal epithelium ( 90 ). In some instances, certain Lactobacillus can aggregate to form a biofilm that is favourable to intestinal cell adhesion and colonization ( 199 ). In line with our findings, Lactiplantibacillus plantarum isolates KA18, KA19, KII1, KII2 and KLAB5 derived from fermented radish displayed a high adherence rate to Caco-2 cells, ranging from 25% to 49%, in contrast to those of Lacticaseibacillus rhamnosus GG, whose adhesion capacity was reported to be 12% ( 186 ). The Caco-2 adhesion rate of Lactiplantibacillus plantarum S2-5 (12 %) ad S4-1 (8.6 %) iolated from Chinese sauerkraut was higher than that of Lacticaseibacillus rhamnosus GG (6.1 %) ( 185 ). In a different study, Lactobacillus johnsonii NCUA063008 and Lactobacillus plantarum sourced from the faeces of wild boars, native pigs and commercial pigs demonstrated a capacity of 20.16% and 17.19% respectively, to Caco-2 cells ( 86 ). However, a higher adhesion capacity to Caco-2 cells than that observed in our study was demonstrated by Lactobacillus reuteri SHA 113 and Lactobacillus johnsonii SHA105 (90%) ( 18 ), and Lactobacillus reuteri LR21 and LR 42 (> 70%) isolated from the poultry gut ( 55 ). Bacterial adhesion is a strain-specific characteristic that is strongly influenced by origin, variations in the expression levels of surface proteins, and environmental factors ( 207 ). Highly adherent strains may serve as good probiotic candidates for imparting beneficial health effects, including immunomodulation and the exclusion of pathogens via competition for binding sites and nutrients in the host intestinal mucosa ( 205 ). In contrast, the isolates Latilactobacillus curvatus (Pb2, banana) and Lactiplantibacillus plantarum (Cu3e, cucumber) demonstrated the lowest adhesion capacities, at 2.60% and 3.58%, respectively (Table 12 ). The reason for this may be attributed to the loss or absence of the S-layer protein from the bacterial surface through chemical means or the shielding of the layer by other molecules during prolonged cultivation ( 229 ). Certain proteins in Lactobacillus species can bind weakly (through noncovalent interactions) to surface components of intestinal cells ( 90 ). Moreover, the EPS layers produced by lactobacilli can block the adhesion of probiotic membrane proteins to the GIT cells ( 122 ). As stated in a previous study, bacterial species with a poor hydrophobic index exhibit decreased cell adhesion ( 203 ). Similarly, Latilactobacillus curvatus JBCC38 and JBCC99 expressed a lower Caco-2 cell adhesion percentage ( 25% recorded for Lacticaseibacillus rhamnosus GG ( 230 ). In contrast to this study, Latilactobacillus curvatus BSF206 demonstrated an adhesion percentage of 13.2%, which was higher than the percentage of Lacticaseibacillus rhamnosus GG (6.1%) ( 215 ). The ability of Latilactobacillus curvatus DN3 to adhere to Caco-2 cells was 16% greater than that of Lacticaseibacillus rhamnosus GG at 7% ( 231 ). Discrepancies in the adhesion capacity of Lactobacillus strain to the host may be due to differences in transpeptidase-dependent proteins, and the specific binding of bacterial surface proteins, pili, peptidoglycans, and lipopolysaccharides to surface receptors of epithelial cells ( 215 ). Probiotic strains with poor adhesion capacity may be unable to effectively adhere to and colonize the host mucosal surfaces to confer health benefits to the host ( 183 ). CPP of Lactobacillus isolates The CPP is an important criterion for the validation of the characteristics of probiotic microorganisms ( 232 ). In this study, the CPP of three isolates, Lactiplantibacillus plantarum (To3a and To3d, tomato) and Latilactobacillus curvatus (Cu2f, cucumber), was higher (87.50%) (Table 13 ) than commercially available probiotic preparations, whose probiotic score is in a range of 75–85% ( 36 , 233 ). Conversely, a high CPP of 100% was recorded for both Lactiplantibacillus plantarum LMEM1002 and LMEM1006 sourced from idli batter samples ( 234 ). Lactiplantibacillus plantarum NKUST 817, 828, and 851 isolated from fermented food ( 233 ) and Lactobacillus johnsonii PUMSKGRI, isolated from a homemade fermented food product, all had a CPP of 91.67% ( 36 ). Notably, these two isolates, Lactiplantibacillus plantarum (To3a and To3d, tomato), met the standard for a good probiotic considering their probiotic characteristics, especially acid and bile tolerance ability. Conclusions This research study presented eighteen Lactobacillus isolates sourced from different food sources. Lactobacillus plantarum To3a and To3d isolates demonstrated promising candidates for various probiotic applications, based on their tolerance to acid and bile salt, cell adhesion capacity, bile salt and cholesterol-lowering ability, and strong inhibition towards pathogenic bacteria. In addition, results on safety aspects revealed that they are safe for human consumption when used as probiotics. The cumulative probiotic potential of To3a and To3d isolates suggests that they may represent a good probiotic candidate essential for the development of novel local functional food products with health-promoting effects. Nevertheless, further comprehensive studies on the technological properties, as well as in vivo studies, should be considered to gain a better insight into their beneficial attributes. Declarations Data availability The datasets generated and/or analyzed during this study are available from the corresponding author upon reasonable request. Acknowledgements The authors would like to thank the National Research Foundation (NRF) of South Africa for their financial support (Grant number: 129095). We would also like to thank the Department of Life and Consumer Sciences, University of South Africa, for their support in providing laboratory supplies. Funding This work is based on research supported by the National Research Foundation (NRF) of South Africa (Grant number: 129095). Authors information Authors and affiliations Department of Life and Consumer Sciences,University of South Africa, Cnr Christiaan de Wet Road and Pioneer Avenue, Florida, Roodepoort 1710, South Africa Chioma Vivian Asiegbu Department of Life and Consumer Sciences,University of South Africa, Cnr Christiaan de Wet Road and Pioneer Avenue, Florida, Roodepoort 1710, South Africa Frederick Tawi Tabit Contributions ACV and FTT participated in the conceptualization and design of the study; ACV drafted the original manuscript; ACV collected the sample and data; ACV performed the experiments; ACV and FTT analyzed the data; FTT supervised; ACV and FTT wrote- reviewed & edited; FTT read and approved the final manuscript. Corresponding author Correspondence to Frederick Tawi Tabit Ethics approval and consent to participate This research protocol was approved by the College of Agriculture and Environmental Science (CAES) Health Research Ethics Committee at the University of South Africa. 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Tables Table 1 : Lactobacillus species identified from chicken gut, tomatoes, cucumber and banana Food source Frequency of Lactobacillus species identified Frequency Chicken gut Lactobacillus johnsonii Limosilactobacillus reuteri Latilactobacillus curvatus Latilactobacillus sakei (n=7; 38.9%) (n=2; 11.1%) (n=2; 11.1%) (n=1; 5.6%) Tomato Lactiplantibacillus plantarum (n=3; 16.6%) Cucumber Lactiplantibacillus plantarum Latilactobacillus curvatus (n=1; 5.6%) (n=1; 5.6%) Banana Latilactobacillus curvatus (n=1; 5.6%) Table 2 : Identification of Lactobacillus species isolated from chicken gut, tomato, cucumber, and banana using 16S rRNA Gene Sequencing Lactobacillus isolates Source Molecular identity Similarity index Mcya Chicken gut Lactobacillus johnsonii 100% Mcyb Chicken gut Lactobacillus johnsonii 100% Mcyc Chicken gut Limosilactobacillus reuteri 100% Mcxb Chicken gut Lactobacillus johnsonii 100% Mcxc Chicken gut Lactobacillus johnsonii 100% Chcg Chicken gut Latilactobacillus curvatus 100% Chcb Chicken gut Latilactobacillus curvatus 100% Chcx2 Chicken gut Lactobacillus johnsonii 99.74% Chcx3a Chicken gut Limosilactobacillus reuteri 99.47% Chcx3b Chicken gut Lactobacillus johnsonii 100% Chcx4a Chicken gut Lactobacillus johnsonii 100% Mcg Chicken gut Latilactobacillus sakei 100% To3a Tomato Lactiplantibacillus plantarum 99.54% To3b Tomato Lactiplantibacillus plantarum 100% To3d Tomato Lactiplantibacillus plantarum 100% Cu2f Cucumber Latilactobacillus curvatus 99.20% Cu3e Cucumber Lactiplantibacillus plantarum 99.73% Pb2 Banana Latilactobacillus curvatus 99.74% Table 3: The acid tolerance level of Lactobacillus isolates after exposure to pH 2.5 and pH 3.0 for 3h Isolates code Cell survival rate (%) at pH 2.5 Cell survival rate (%) at pH 3 ¥ To3a 78.39 0.69 ef 97.20 5.79 g To3b 67.82 6.03 cd 80.74 7.81 de To3d 73.85 0.84 de 84.00 1.57 ef Chx3a 83.83 6.06 fg 89.87 1.88 efg Chx3b 64.46 0.92 c 71.17 7.24 bc Cu2f 44.99 1.55 a 53.79 2.19 a ¥ Cu3e 68.59 3.56 cd 89.9 3.39 fg ¥ Pb2 72.39 6.25 de 89.57 7.47 fg Mcyc 89.94 6.68 g 98.1 1.32 g Mcxb 53.35 1.75 b 65.65 2.48 b Mcxc 63.84 6.12 c 75.71 0.75 cd ¥ GG 67.65 1.14 cd 93.03 5.37 g The data shows the mean ± standard deviation of triplicate values of independent analyzes. Columns with different superscript letters indicate significant differences. ¥ = Lactobacillus isolates for which the survival rate was significantly different and higher at pH 2.5 and 3.0 Table 4: The bile salt tolerance level of Lactobacillus isolates after exposure to 0.3% and 0.5% bile salt for 4 h Isolates code Cell survival rate (%) at 0.3% bile salt Cell survival rate (%) at 0.5% bile salt £ To3a 98.72 1.37 e 89.00 0.75 d To3b 98.74 0.78 e 97.42 1.63 e To3d 98.22 1.60 de 94.57 4.03 e Chx3a 89.66 5.93 b 73.64 1.97 b Chx3b 81.12 0.85 a 62.05 1.51 a £ Cu2f 99.08 1.79 e 81.88 1.52 c Cu3e 95.37 3.80 cde 83.20 6.30 cd Pb2 90.87 1.58 bc 86.70 1.31 cd Mcyc 98.37 0.86 de 85.25 0.67 cd Mcxb 99.31 0.93 e 95.43 2.29 e Mcxc 93.38 5.66 bcd 69.64 5.91 b £ GG 92.92 0.97 bc 88.77 3.08 d The data shows mean ± standard deviation of triplicate values of independent analyzes. Columns with different superscript letters indicate significant differences. £ = Lactobacillus isolates for which the survival rate was significantly different and higher at 0.3% and 0.5% bile salt Table 5: The BSH activity, EPS production ability, and cholesterol-lowering ability of Lactobacillus isolates Isolates code BSH activity a EPS producing ability c Cholesterol-lowering ability b To3a ++ + 49 1.27 h To3b + + 40 1.17 ef To3d + + 0.20 i Chx3a ++ + 1.37 j Chx3b + – 0.19 e Cu2f + + 1.58 b Cu3e ++ + 46 1.73 g Pb2 + – 0.98 d Mcyc + – c Mcxb + + d Mcxc – – a GG + + f a BSH activity is shown based on the diameter of precipitation zones: No precipitation (–), precipitation zone (+) up to 10 mm, precipitation zone 10–15 mm (++), and precipitation zone >15 mm (+++). b Cholesterol–lowering ability values are mean ± SD of two independent observations (n=2) of each sample. abcd Means bearing different superscripts in a column differ significantly (P<0.05). c EPS producing ability is displayed based on (+) means positive growth while (–) means negative growth Table 6: DPPH scavenging activity (%) of Lactobacillus isolates Antioxidant assay DPPH scavenging activity (%) To3a 19.73 ±2.28 cd To3b 17.92±1.09 cd € To3d 24.45±1.15 f Chcx3a 18.26±0.63 cd Chcx3b 17.08±1.65 c Cu2f 6.81±1.56 a Cu3e 18.05±2.93 cd Pb2 11.11±2.25 b # Mcyc 20.90±1.62 de Mcxb 7.85±1.36 a Mcxc 13.33±1.66 b GG 23.20±1.35 ef Ascorbic acid (10mg/ml) 35.63±2.20 g Values are mean ± SD of three independent observations (n=3) of each sample. abcd Means bearing different superscripts in a column differ significantly (P<0.05) € = Lactobacillus isolates for which the DPPH scavenging activity was significantly (p≤0.05) higher than that of the reference strain # = Lactobacillus isolates for which the DPPH scavenging activity was not significantly different (p≤0.05) from that of the reference strain Table 7: Antimicrobial activity of CFS of Lactobacillus against pathogenic organisms Isolates code Zone of inhibition a (mm) E . coli S. aureus B . cereus L . monocytogenes ∞ To3a ++ +++ ++ ++ ∞ To3b ++ +++ ++ ++ ∞ To3d ++ ++ ++ ++ Chx3a – – – ++ Chx3b – – ++ – Cu2f + ++ ++ ++ Cu3e – – – – Pb2 – – ++ + Mcyc – – – – Mcxb – – ++ ++ Mcxc – – – – GG ++ ++ ++ ++ a Results of independent experiments (n =3) of inhibition zones: (–) = not detected; (+) = inhibitory zone 15 mm. ∞ = Lactobacillus isolates with antimicrobial activity similar with that of the reference strain (GG), with an inhibition zone of 10 to 15 mm or more against E. coli , S. aureus , B. cereus and L. monocytogenes Table 8: The auto-aggregation ability of Lactobacillus isolates after 2 h and 4 h incubation Isolates code Auto-aggregation (%) 2h 4h µ To3a 24 4.00 bc 4.62 d To3b 22. b 29.33 bcd µ To3d 26.67 bc cd µ Chcx3a b cd µ Chcx3b d 48.67 e µ Cu2f d e Cu3e ab abc Pb2 a a µ Mcyc 33.33 cd e Mcxb ab ab Mcxc bc cd GG ab abcd Values are mean ± SD of three independent observations (n=3) of each sample. abcd Means bearing different superscripts in a column differ significantly (P<0.05). µ = Lactobacillus isolates with a significant (p ≤ 0.05) increase in auto-aggregation (%) from 2 to 4 h incubation and whose auto-aggregation (%) was significantly higher than that of the reference strain after 2 and 3 h incubation, respectively Table 9: Cell surface hydrophobicity ability of Lactobacillus isolates after 1 hr incubation Isolates code Cell surface hydrophobicity (%) To3a 16.00 4.00 b To3b 18.67 bcd To3d d Chcx3a a # Chcx3b e € Cu2f g € Cu3e 72.00 f Pb2 a € Mcyc g Mcxb bc Mcxc cd GG e Values are mean ± SD of three independent observations (n=3) each sample. abcd Means bearing different superscripts in a column differ significantly (P<0.05). € = Lactobacillus isolates for which the cell surface hydrophobicity values were significantly (p≤0.05) higher than those of the reference strain. # = Lactobacillus isolates for which the cell surface hydrophobicity values were not significantly different (p>0.05) from that of the reference strain Table 10: Antibiotic susceptibility profile of Lactobacillus isolates using the disk diffusion method Antibiotics Susceptibility profile of Lactobacillus i solates To3a To3b To3d Chx3a Chx3b Cu2f Cu3e Pb2 Mcyc Mcxb Mcxc GG Penicillin G (10 μg) S S S S S S S S S S S S Chloramphenicol (30 μg) S S S S S S S S S S S S Kanamycin (30 μg) R R R R R R R R R R R R Gentamycin (10 μg) R R R R R R R R R R R R Streptomycin (10 μg) R R R R R R R R R R R R Tetracycline (30 μg) I S S R R S S R R R R S Erythromycin (15 μg) S S S S S S S S S S S S Ampicillin (10 μg) S S S S S S S S S S S S Vancomycin (30 μg) R R R I S R R R R S S R Results are presented as S: Susceptible (≥21 mm), I: intermediate (16–20 mm), R: resistance (≤ 15 mm) Table 11: Blood haemolysis and DNase activity of Lactobacillus isolates Isolates code Haemolysis a DNase activity b To3a ϒ –ve To3b ϒ –ve To3d ϒ –ve Chx3a ϒ –ve Chx3b ϒ –ve Cu2f ϒ –ve Cu3e ϒ –ve Pb2 ϒ –ve Mcya ϒ –ve Mcxb ϒ –ve Mcxc ϒ –ve GG ϒ –ve a ϒ: means no haemolysis, β: means complete haemolysis, α: means partial haemolysis. b –ve indicates no DNase activity, +ve indicates DNase activity Table 12: Adhesion capacity of Lactobacillus isolates to Caco-2 epithelial cells Isolates code Adhesion (%) To3a 9.94 .06 de To3b 6.85 bc To3d g Chcx3a c Chcx3b h Cu2f b Cu3e 3.58 a Pb2 2.60 0.83 a Mcyc ef Mcxb b Mcxc d GG f Values are mean ± SD of three independent observations (n=3). abcd Means bearing different superscripts in a column differ significantly (P<0.05) Table 13: Cumulative probiotic potential (CPP) of Lactobacillus isolates Indicator score To3a To3b To3d Chcx3a Chcx3b Cu2f Cu3e Pb2 Mcyc Mcxb Mcxc Probiotic characters Acid tolerance % ≥ 50 = 1 % < 50 = 0 1 1 1 1 1 0 1 1 1 1 1 Bile salt tolerance % ≥ 50 = 1 % 30 = 1 1 0 1 1 1 1 0 0 1 0 1 Cell surface hydrophobicity % ≤ 30 = 0 % > 30 = 1 0 0 0 0 1 1 0 0 1 0 0 Antimicrobial activity < 15 mm = 0 ≥ 15 mm = 1 1 1 1 0 0 1 0 0 0 0 0 Safety parameters Tetracycline antibiotic susceptibility Sensitive/ intrinsic resistant=0 Resistant=1 1 1 1 0 0 1 1 0 0 0 0 Haemolytic activity Activity=0 No activity=1 1 1 1 1 1 1 1 1 1 1 1 DNase activity Activity=0 No activity=1 1 1 1 1 1 1 1 1 1 1 1 Total score 7/8 6/8 7/8 5/8 6/8 7/8 5/8 4/8 6/8 4/8 5/8 Cumulative probiotic potential 87.50% 75% 87.50% 62. 50 % 75 % 87.50% 62. 50% 50 % 75 % 50 % 62. 50 % Additional Declarations No competing interests reported. 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1","display":"","copyAsset":false,"role":"figure","size":275658,"visible":true,"origin":"","legend":"\u003cp\u003ePhylogenetic analysis of \u003cem\u003eLactobacillus\u003c/em\u003e strains isolated from chicken intestine, tomatoes, cucumber and banana.\u003c/p\u003e","description":"","filename":"Figure1.png","url":"https://assets-eu.researchsquare.com/files/rs-7594770/v1/4a7338d862be22aad6c52684.png"},{"id":93808112,"identity":"414bae33-4001-4e15-9424-b05515000e8f","added_by":"auto","created_at":"2025-10-17 18:52:12","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":215280,"visible":true,"origin":"","legend":"\u003cp\u003ePhylogenetic analysis of \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e and \u003cem\u003eLimosilactobacillus\u003c/em\u003e \u003cem\u003ereuteri\u003c/em\u003estrains from the chicken intestine.\u003c/p\u003e","description":"","filename":"Figure2.png","url":"https://assets-eu.researchsquare.com/files/rs-7594770/v1/051ae94f8d9bf6af963ecd1b.png"},{"id":95312072,"identity":"a3dc9a9b-45fa-4128-8965-437c49f6b69d","added_by":"auto","created_at":"2025-11-06 15:46:41","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3242617,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7594770/v1/94669473-8fc0-47be-a6de-2c8c0b48db8d.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Prospecting and characterization of potential probiotic Lactobacillus species from animal gut and food sources","fulltext":[{"header":"Background","content":"\u003cp\u003e\u003cem\u003eLactobacillus\u003c/em\u003e is a prominent member of lactic acid bacteria (LAB), which are Gram-positive, rod-shaped, microaerophilic \u003cem\u003ebacilli\u003c/em\u003e, with over 290 species having catalase-and oxidase-negative features (\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e). They include, among others, \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e, \u003cem\u003eLactobacillus acidophilus\u003c/em\u003e, \u003cem\u003eLacticaseibacillus casei\u003c/em\u003e, \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e, \u003cem\u003eLimosilactobacillus fermentum\u003c/em\u003e, \u003cem\u003eLevilactobacillus brevis\u003c/em\u003e, \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e, \u003cem\u003eLigilactobacillus salivarius\u003c/em\u003e, and \u003cem\u003eLacticaseibacillus paracasei\u003c/em\u003e (\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e). \u003cem\u003eLactobacillus\u003c/em\u003e species are typically found in diverse niches, including the gastrointestinal tract (GIT) of humans and animals, plant surfaces, fermented foods, water, wine, soil, and sewage, among others (\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e, \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e). These bacteria play a considerable role in food fermentation, flavour, texture, and aroma development in food products (\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e), and they are \"generally recognized as safe\"(GRAS) and have a qualified safety estimate (QPS) status (\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e). Hence, it may be safely exploited for extensive probiotic application.\u003c/p\u003e\u003cp\u003eSome \u003cem\u003eLactobacillus\u003c/em\u003e species are probiotics, live microorganisms, which, when consumed in sufficient amounts, benefit the host's health (\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e). The health benefits of probiotics, which include intestinal microbiota balance, intestinal tract relief, allergy alleviation, obesity, diabetes, bacterial vaginosis, and oral diseases, have been reported in many studies (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e, \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e). In addition to those mentioned above, other health advantages, such as the prevention and management of cancer, hypercholesterolemia, lactose intolerance, improved immune response, inflammatory bowel disease, and irritable bowel syndrome, have been reported (\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e). Consequently, these valuable microbes have gained extensive recognition and research interest over the years due to their broad spectrum of positive health effects (\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eMany probiotic strains have been successfully isolated from fruits and vegetables (\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e), fruits and fermented foods (\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e), leaves of food plants (\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e), chickens (\u003cspan additionalcitationids=\"CR18 CR19\" citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e), the GIT of snakehead fish (\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e), the GIT of wild boar (\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e), and the intestinal mucosa of healthy piglets (\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e). For example, \u003cem\u003eLactobacillus acidophilus\u003c/em\u003e CM1 and \u003cem\u003eLactobacillus delbrueckii\u003c/em\u003e OS1 strains isolated from different food sources (\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e) and \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e strains isolated from silages from different forage plants, artisanal salami, and fermented cocoa beans demonstrated promising probiotic properties (\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e). Notably, the isolation of these strains from various food sources has great potential because of their naturalness as well as the techno-functional and health benefits they may possess (\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eWhile probiotics offer well-documented health benefits, accessibility and affordability remain significant challenges in developing countries, especially in rural areas (\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e, \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e). Most commercially available probiotic products in Africa are based on \u003cem\u003eLactobacillus\u003c/em\u003e strains sourced from countries outside the continent, rather than utilizing indigenous species from the African microbiota (\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e), with bacterial strains such as \u003cem\u003eLacticaseibacillus paracasei\u003c/em\u003e Shirota, \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG, and \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e RC-14 recognized as the most commercially available and established probiotics (\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e, \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e, \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e). Unfortunately, some of these imported probiotic cultures are expensive for many African locals (\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e), thus, hampering the development of innovative food products from locally sourced probiotic organisms. In response to the limited representation of locally sourced probiotic strains in commercial formulations, this study aimed to prospect, isolate, and characterize \u003cem\u003eLactobacillus\u003c/em\u003e species from animal GIT, as well as from fruit and vegetable sources, within the Johannesburg metropolitan area of South Africa.\u003c/p\u003e"},{"header":"Materials and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003eSample collection\u003c/h2\u003e\u003cp\u003eFruit (bananas) and vegetables (tomatoes and cucumbers) were purchased from local street stalls and retail stores in the central business district (CBD) of Johannesburg, Gauteng Province, South Africa. In contrast, fresh chicken guts were purchased from live poultry stores in the CBD of Johannesburg, Gauteng Province, South Africa. The samples were aseptically collected in sterile plastic containers. They were immediately transported in a cooler ice box to the microbiology laboratory for further analysis within 24 h.\u003c/p\u003e\u003cp\u003e\u003cb\u003eEnrichment and isolation of\u003c/b\u003e \u003cb\u003eLactobacillus\u003c/b\u003e \u003cb\u003especies\u003c/b\u003e\u003c/p\u003e\u003cp\u003eFruit and vegetable samples were washed separately with sterile distilled water. Banana fruit (n\u0026thinsp;=\u0026thinsp;5) or vegetable samples (n\u0026thinsp;=\u0026thinsp;5) were aseptically cut on sterile aluminium foil into separate small pieces, after which, one gram (1 g) from each dissected fruit or vegetable sample was separately homogenized in 9 milliliter (ml) of 0.1% sterile peptone solution (Merck, SA) using a sterile Waring laboratory blender for 2 min.\u003c/p\u003e\u003cp\u003eBroiler chicken caecal (n\u0026thinsp;=\u0026thinsp;3) and small intestine (n\u0026thinsp;=\u0026thinsp;3) portions were aseptically dissected separately to obtain the contents inside. Then, 1 g of each caecal or small intestine content was separately homogenized in 9 ml of peptone solution.\u003c/p\u003e\u003cp\u003eFollowing homogenization, each sample was serially diluted up to 10⁷\u0026ndash;fold with buffered peptone water. Subsequently, 1 ml from each dilution was aseptically spread onto De Man, Rogosa, and Sharpe (MRS) agar plates (Merck, South Africa) and incubated anaerobically at 37\u0026deg;C for 24 to 48 h. Thereafter, a single colony of each \u003cem\u003eLactobacillus\u003c/em\u003e isolate, showing different morphologies from each other, was carefully picked and streaked onto a newly prepared MRS agar plate to obtain pure bacterial cultures. These cultures were then harvested and stored at \u0026ndash; 80\u0026deg;C in MRS broth containing 20% sterile glycerol (v/v) for further analyzes.\u003c/p\u003e\u003c/div\u003e\n\u003ch3\u003eIdentification of lactobacilli by 16S rRNA gene sequencing\u003c/h3\u003e\n\u003cp\u003e\u003cem\u003eLactobacillus\u003c/em\u003e species isolates were subjected to Gram staining, motility, and catalase tests as described by Hidayat et al. (\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e). Motile, catalase-negative, Gram-positive, rod-shaped isolates were subjected to molecular characterization.\u003c/p\u003e\u003cp\u003eGenomic DNA was extracted using the ZR Fungal/Bacterial DNA Miniprep\u0026trade; Kit (Zymo Research, Irvine, USA). The DNA was quantified using a NanoDrop\u0026trade; One UV-Vis Spectrophotometer (Thermo Fisher Scientific, USA). The 16S rRNA genes of the isolates were amplified using a set of universal primers 27F (5\u0026rsquo;-AGAGTTTGATCCTGGCTCTCAG-3\u0026rsquo;) and 1429R (5\u0026rsquo;-GGTTACCTTGTTACGACTT-3\u0026rsquo;). The polymerase chain reaction (PCR) was performed in a 25 \u0026micro;l volume, which consisted of 12.5 \u0026micro;l of 2x PCR master mix (Anatech, SA), 1 \u0026micro;l each of the forward and reverse primers, 1 \u0026micro;l of chromosomal DNA, and nuclease-free water to make up the final volume. PCR amplification was performed using Bio-Rad T100 \u003csup\u003eTM\u003c/sup\u003e Thermocycler under the following program: pre-denaturation for 3 min at 95\u0026deg;C, followed by 35 cycles of denaturation (95\u0026deg;C, 30 s), annealing (55\u0026deg;C, 55 s), extension (72\u0026deg;C, 60 s) and final extension at 72\u0026deg;C for 10 min. The quality of the PCR product was examined on a 1.5% (w/v) agarose gel using the Mini-Sub\u0026reg; Cell GT (Bio-Rad Laboratories Inc., California, USA) and visualized using the Gel DOC\u0026trade; XR\u0026thinsp;+\u0026thinsp;Imaging system after staining with ethidium bromide. PCR purification and sequencing of the amplified PCR products were conducted by Inqaba Biotechnology Company, Pretoria, SA. The forward and reverse 16S rRNA gene sequences of the isolates were edited using BioEdit Sequence Alignment Editor version 7.2 software. The identity of the bacterial strains was determined using the Basic Logical Alignment Search Tool (BLAST) program (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://blast.ncbi.nlm.nih.gov/Blast.cgi\u003c/span\u003e\u003cspan address=\"https://blast.ncbi.nlm.nih.gov/Blast.cgi\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e).\u003c/p\u003e\n\u003ch3\u003eScreening for Probiotic properties of bacterial isolates\u003c/h3\u003e\n\u003cp\u003e\u003cb\u003ePreparation of\u003c/b\u003e \u003cb\u003eLactobacillus\u003c/b\u003e \u003cb\u003ecell suspensions\u003c/b\u003e\u003c/p\u003e\u003cp\u003ePure cultures of each \u003cem\u003eLactobacillus\u003c/em\u003e species from MRS agar plates were inoculated into 10 ml MRS broth and incubated at 37\u0026deg;C for 18 h to obtain a pure culture. One ml of each culture was centrifuged at 10000 rpm for 5 min at 4\u0026deg;C. The cell pellets were washed twice with phosphate-buffered saline (PBS) at pH 7.2, and the washed pellets were used to prepare cell cultures for subsequent analysis. For each experimental assay, the optical density (OD) at 600 nm of the bacterial suspensions was adjusted to either 1.0 (for 2,2-diphenyl-1-picrylhydrazyl scavenging and cell adhesion activity) or 0.25 (for antimicrobial activity, cell auto-aggregation, cell surface hydrophobicity, antibiotic susceptibility, and DNase activity) using a Genesys 10S VIS spectrophotometer (Thermo Scientific, USA).\u003c/p\u003e\n\u003ch3\u003eAcid tolerance test\u003c/h3\u003e\n\u003cp\u003eEach pure culture of \u003cem\u003eLactobacillus\u003c/em\u003e was adjusted to 10\u003csup\u003e9\u003c/sup\u003e CFU/ml and used for analyzes. The acid tolerance of the selected \u003cem\u003eLactobacillus\u003c/em\u003e isolates was examined by the method described by Kumar et al. (\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e). The acidic medium was prepared by adjusting the MRS broth with 1 N hydrochloric acid (HCl) (Sigma, SA) to a pH of 2.5 or 3.0. Approximately 0.1 ml of each \u003cem\u003eLactobacillus\u003c/em\u003e cell suspension at 10^9 CFU/ml was inoculated into 10 ml of acidic MRS broth at pH 2.5 and pH 3 and incubated anaerobically at 37\u0026deg;C for 0 and 3 h. Next, 0.1 ml of each of the serial dilutions (10\u003csup\u003e1\u003c/sup\u003e \u0026ndash; 10\u003csup\u003e6\u003c/sup\u003e) of the acid medium culture was spread on an MRS agar plate, followed by anaerobic incubation for 48 h at 37\u0026deg;C. The survival rate of \u003cem\u003eLactobacillus\u003c/em\u003e isolates under low acidic conditions was calculated using the formula described by Dehghani et al. (\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e). \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG (Anatech, SA) was used as a positive control.\u003c/p\u003e\u003cp\u003eSurvival rate % = \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\frac{\\text{l}\\text{o}\\text{g}\\text{C}\\text{F}\\text{U}\\:\\text{o}\\text{f}\\:\\text{v}\\text{i}\\text{a}\\text{b}\\text{l}\\text{e}\\:\\text{c}\\text{e}\\text{l}\\text{l}\\text{s}\\:\\text{s}\\text{u}\\text{r}\\text{v}\\text{i}\\text{v}\\text{e}\\text{d}\\:}{\\text{l}\\text{o}\\text{g}\\text{C}\\text{F}\\text{U}\\:\\text{o}\\text{f}\\:\\text{i}\\text{n}\\text{i}\\text{t}\\text{i}\\text{a}\\text{l}\\:\\text{c}\\text{e}\\text{l}\\text{l}\\text{s}\\:\\text{i}\\text{n}\\text{n}\\text{o}\\text{c}\\text{u}\\text{l}\\text{a}\\text{t}\\text{e}\\text{d}}\\)\u003c/span\u003e\u003c/span\u003e \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\times\\:\\)\u003c/span\u003e\u003c/span\u003e 100\u003c/p\u003e\n\u003ch3\u003eBile tolerance test\u003c/h3\u003e\n\u003cp\u003eWith minor modifications, the method described by Kumar et al. (\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e) was used to test the bile salt tolerance of the \u003cem\u003eLactobacillus\u003c/em\u003e isolates. Approximately 0.1 ml of each \u003cem\u003eLactobacillus\u003c/em\u003e cell suspension at 10\u003csup\u003e9\u003c/sup\u003e CFU/ml was inoculated into 10 ml of MRS broth containing 0.3% or 0.5% (w/v) bile salt (Sigma-Aldrich) and incubated at 37\u0026deg;C for 0 and 4 h under anaerobic conditions. After incubation, 0.1 ml of each serial dilution (10\u003csup\u003e1\u003c/sup\u003e \u0026ndash; 10\u003csup\u003e6\u003c/sup\u003e) was spread on MRS agar plates and incubated anaerobically at 37\u0026deg;C for 48 h. The survival rate was calculated using the formula applied for the acid tolerance. \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG was used as a positive control.\u003c/p\u003e\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\u003ch2\u003eBile salt hydrolase (BSH) assay\u003c/h2\u003e\u003cp\u003eThe BSH activity of the isolates was tested qualitatively using the direct plate BSH method described by Habib et al. (\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e). MRS agar plates supplemented with calcium chloride (CaCl\u003csub\u003e2\u003c/sub\u003e) (0.37 g/L) (Merck, SA) and sodium taurocholate (TDCA) (0.5%, w/v) (Merck, SA) were freshly prepared, and wells were created on the agar plates with sterile blue P1000 pipette tips (SBPT). Thereafter, 0.1 ml of each \u003cem\u003eLactobacillus\u003c/em\u003e culture was pipetted into the wells of the supplemented media plates and incubated anaerobically at 37\u0026deg;C for 48 h. Agar plates without CaCl\u003csub\u003e2\u003c/sub\u003e and TDCA were used as controls. The precipitation of bile acids around the wells indicated BSH activity. BSH activity was expressed based on the diameter of the precipitation zones around the wells. It was evaluated as follows: no precipitation (\u0026minus;), precipitation zone (+) up to 10 mm, precipitation zone 10\u0026ndash;15 mm (++), and precipitation zone\u0026thinsp;\u0026gt;\u0026thinsp;15 mm (+++).\u003c/p\u003e\u003cp\u003e\u003cb\u003eCholesterol-lowering ability of\u003c/b\u003e \u003cb\u003eLactobacillus\u003c/b\u003e \u003cb\u003eisolates\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe in vitro \u003cem\u003echolesterol\u003c/em\u003e-lowering ability assay was conducted using the \u003cem\u003eo\u003c/em\u003e-phthalaldehyde method detailed by Srinivash et al. (\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e). A syringe filter (0. 45 \u0026micro;m) was used to filter water-soluble cholesterol (10 mg/100 ml; Merck, SA). Then,100 \u0026micro;l of this water-soluble cholesterol was mixed with 9.9 ml of MRS broth supplemented with 0.3% bile salt. The mixture was inoculated with 1 ml of a suspension (10^8 CFU/ml) of each \u003cem\u003eLactobacillus\u003c/em\u003e isolate and incubated at 37\u0026deg;C for 24 h. The MRS broth without 0.3% bile salt and water-soluble cholesterol served as the control medium. After incubation, the cells were pelleted by centrifugation at 8000 \u0026times; g for 10 min. One ml of the supernatant was added to a sterile test tube, along with 1 ml of potassium hydroxide (33% w/v; Merck, SA) and 2 ml of 95% absolute ethanol. The mixture was vortexed for 1 min, incubated at 35\u0026deg;C for 15 min, and then placed in a water bath at 37\u0026deg;C for an additional 15 min. After cooling to room temperature, 2 ml of distilled water and 3 ml of hexane (Sigma, SA) were added, and the mixture was vortexed for 1 min. The mixture was then allowed to stand for 15 min to facilitate phase separation. One ml of the hexane layer was transferred into a sterile glass vial and heated in a water bath at 65\u0026deg;C to evaporate the hexane solvent. After hexane evaporation, 2 ml of o-phthalaldehyde reagent (Merck, SA) was added to the residue. After thorough mixing, 0.5 ml of concentrated sulfuric acid (Sigma, SA) was carefully added, and the mixture was vortexed for 1 min. The resulting mixture was allowed to stand for 10 min at room temperature before the absorbance at 550 nm was measured. Cholesterol reduction (%) was calculated as: 1 \u0026ndash; (absorbance of culture supernatant 550/absorbance of control 550) \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\times\\:\\)\u003c/span\u003e\u003c/span\u003e 100\u003c/p\u003e\u003cp\u003e\u003cb\u003eExopolysaccharide (EPS) producing ability of\u003c/b\u003e \u003cb\u003eLactobacillus\u003c/b\u003e \u003cb\u003eisolates\u003c/b\u003e\u003c/p\u003e\u003cp\u003ePure cultures of the samples were evaluated according to the method of Abouloifa et al. (\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e), for the ability of \u003cem\u003eLactobacillus\u003c/em\u003e species to produce EPS. Freshly cultivated isolates were streaked on ruthenium red milk agar plates containing 10% (w/v) skim milk (Merck, SA), 1% (w/v) sucrose (Sigma, SA), 0.08% (w/v) ruthenium red (Sigma, SA), and 1.5% (w/v) agar (Merck, SA). After anaerobic incubation for 48 h at 37\u0026deg;C, white colonies indicated a positive result for EPS production, whereas pink colonies indicated a negative outcome.\u003c/p\u003e\u003cp\u003e\u003cb\u003e2,2-Diphenyl-1-picrylhydrazyl (DPPH) scavenging activity\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe method proposed by Kim et al. (\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e) was used with slight modifications to determine the DPPH radical scavenging ability of the isolates. A DPPH radical solution was prepared by dissolving 4 mg of DPPH in 100 ml of methanol in a glass tube wrapped in aluminum foil. Two milliliter of DPPH solution was added to 1 ml of each bacterial suspension (OD600 nm\u0026thinsp;=\u0026thinsp;1.0) and mixed thoroughly. The resulting mixture was kept in the dark at room temperature for 30 min. A mixture of 2 ml of DPPH and 2 ml of methanol was used as a negative control, while ascorbic acid (10 mg/ml) (Merck, SA) served as the positive control. The assay was performed in triplicate, and the absorbance (OD) of the resulting solution was measured at 517 nm.\u003c/p\u003e\u003cp\u003eThe scavenging effect (%) = (Ac\u0026ndash;As)/Ac \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\times\\:\\)\u003c/span\u003e\u003c/span\u003e 100, where As is the absorbance of the test sample, and Ac is the absorbance of the control sample.\u003c/p\u003e\u003c/div\u003e\n\u003ch3\u003eAntimicrobial activity against pathogenic bacteria\u003c/h3\u003e\n\u003cp\u003eThe agar well diffusion method described by Sakandar et al.(\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e) was used to examine the antimicrobial activity of \u003cem\u003eLactobacillus\u003c/em\u003e isolates against pathogenic bacteria. The test pathogens used in this study were \u003cem\u003eStaphylococcus aureus\u003c/em\u003e (\u003cem\u003eS. aureus\u003c/em\u003e) ATCC 25923, \u003cem\u003eEscherichia coli\u003c/em\u003e (\u003cem\u003eE. coli\u003c/em\u003e) ATCC 25922, \u003cem\u003eListeria monocytogenes\u003c/em\u003e (\u003cem\u003eL. monocytogenes\u003c/em\u003e) ATCC 15313, and \u003cem\u003eBacillus cereus\u003c/em\u003e (\u003cem\u003eB. cereus\u003c/em\u003e) ATCC 11778 (Anatech, SA). \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG served as a probiotic reference strain. The cell-free supernatant (CFS) of each isolate was prepared by incubating 10 ml of MRS broth at pH 7.2 overnight at 37\u0026deg;C, followed by centrifugation at 10,000 rpm for 5 min at 4\u0026deg;C. The CFS was collected using a sterile syringe with a needle. The CFS was then sterilized using a 0.22 \u0026micro;m filter (Anatech, SA). Each pathogenic strain was grown overnight at 37\u0026deg;C in nutrient broth (Sigma, SA) and adjusted to an OD of 0.25 at 600 nm. Approximately 0.1 ml of each pathogen cell suspension was spread on Muller-Hinton agar (MHA) (Merck, SA) using a sterile cotton swab. The agar plates were allowed to dry for 1\u0026ndash;2 min before wells were created on the MHA plate using the SBPT. The CFS of the isolates were pipetted into each agar well, and the plate was incubated at 37\u0026deg;C for 16\u0026ndash;24 h. Clear zones around each agar well were measured in millimeter (mm).\u003c/p\u003e\u003cp\u003e\u003cb\u003eCell auto\u003c/b\u003e-\u003cb\u003eaggregation\u003c/b\u003e\u003c/p\u003e\u003cp\u003eFour ml of each bacterial suspension (OD600 nm\u0026thinsp;=\u0026thinsp;0.25) was vortexed for 10 s, after which the suspension was incubated at 37\u0026deg;C for 2 and 4 h. An aliquot of 1 ml of the upper suspension was taken using a sterile Pasteur pipette, and its absorbance was determined at an OD of 600 nm. Auto-aggregation (%) = [(1\u0026ndash; (A\u003csub\u003et\u003c/sub\u003e/A\u003csub\u003e0\u003c/sub\u003e)] \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\times\\:\\)\u003c/span\u003e\u003c/span\u003e 100, where \u003cb\u003eA\u003c/b\u003e\u003csub\u003e\u003cb\u003et\u003c/b\u003e\u003c/sub\u003e is the absorbance at time t\u0026thinsp;=\u0026thinsp;2 or 4 h, while \u003cb\u003eA\u003c/b\u003e\u003csub\u003e\u003cb\u003eo\u003c/b\u003e\u003c/sub\u003e is the absorbance at time t\u0026thinsp;=\u0026thinsp;0 h (\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e).\u003c/p\u003e\n\u003ch3\u003eCell surface hydrophobicity\u003c/h3\u003e\n\u003cp\u003eThe cell surface hydrophobicity was determined using the method described by Barache et al. (\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e). The initial absorbance of the bacterial cell suspension (A\u003csub\u003e0\u003c/sub\u003e) at 600 nm was recorded. One ml of xylene was added to three ml of each cell suspension (OD600 nm\u0026thinsp;=\u0026thinsp;0.25), and mixed by vortexing for 2 min. The bacterial suspension was incubated at 37\u0026deg;C for 1 h to allow phase separation. The lower aqueous phase was carefully removed, and its absorbance (A\u003csub\u003et\u003c/sub\u003e) at OD 600nm was measured. Cell surface hydrophobicity (%) = [(1\u0026ndash; (A\u003csub\u003et\u003c/sub\u003e/A\u003csub\u003e0\u003c/sub\u003e)] \u0026times; 100, where A\u003csub\u003et\u003c/sub\u003e is the absorbance of the aqueous phase and A\u003csub\u003e0\u003c/sub\u003e is the absorbance of the initial bacterial suspension.\u003c/p\u003e\u003cp\u003e\u003cb\u003eSafety evaluation of\u003c/b\u003e \u003cb\u003eLactobacillus\u003c/b\u003e \u003cb\u003eisolates\u003c/b\u003e\u003c/p\u003e\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\u003ch2\u003eAntibiotic susceptibility\u003c/h2\u003e\u003cp\u003eThe disk diffusion method described by Mohammad et al. (\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e) was used in this study to assess the antibiotic susceptibility of the isolates. Approximately 0.1 ml of each bacterial cell suspension (OD600 nm\u0026thinsp;=\u0026thinsp;0.25) was spread evenly on the surface of the MRS agar plate and allowed to dry for approximately 8\u0026ndash;15 min. Subsequently, different antibiotic discs (Separations, SA) containing ampicillin (10 \u0026micro;g), erythromycin (15 \u0026micro;g), chloramphenicol (30 \u0026micro;g), tetracycline (30 \u0026micro;g), gentamycin (10 \u0026micro;g), streptomycin (10 \u0026micro;g), vancomycin (30 \u0026micro;g) penicillin G (10 \u0026micro;g) and kanamycin (30 \u0026micro;g) were placed on the surface of the inoculated agar plates aseptically with sterile forceps. After incubation at 37\u0026deg;C for 24 h, the diameters of the inhibition zones were measured using a ruler in mm. The inhibition zones were categorized as sensitive (\u0026ge;\u0026thinsp;21 mm), intermediate (16\u0026ndash;20 mm), or resistant (\u0026le;\u0026thinsp;15 mm), as detailed by Reuben et al. (\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\u003ch2\u003eHaemolytic activity\u003c/h2\u003e\u003cp\u003eEach isolate was streaked onto Columbia agar (Anatech, SA) plates supplemented with 5% (w/v) sterile sheep blood. After incubation at 37\u0026deg;C for 24\u0026ndash;48 h, the plates were examined for signs of haemolysis: β-haemolysis (clear zones around colonies), α-haemolysis (green-hued zones around colonies), or ϒ-haemolysis (no visible zones around colonies) (\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec13\" class=\"Section2\"\u003e\u003ch2\u003eDeoxyribonuclease (DNase) activity\u003c/h2\u003e\u003cp\u003eIsolates were tested for DNase enzyme production by streaking each culture (OD600 nm\u0026thinsp;=\u0026thinsp;0.25) onto DNase agar (Separations, SA) plates. After incubation at 37\u0026deg;C for 24\u0026ndash;48 h, the plates were flooded with 1 M HCl. A clear zone around the colonies was considered positive for DNase production (\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e). \u003cem\u003eS. aureus ATCC 25923\u003c/em\u003e was used as a positive control.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e\u003ch2\u003eCell adhesion assay\u003c/h2\u003e\u003cp\u003eThe human colon adenocarcinoma cell line (Caco-2 cells) (CELLONEX, SA) was kindly provided by Dr. Wale Oladipo (Department of Life Sciences, University of South Africa). Caco-2 cells were grown in a humidified incubator of 5% CO\u003csub\u003e2\u003c/sub\u003e at 37\u0026deg;C in Dulbecco\u0026rsquo;s modified Eagle medium (DMEM) (Sigma-Aldrich, SA) supplemented with 10% inactivated fetal bovine serum (Sigma-Aldrich, SA), 100 units/ml penicillin (Thermo Fisher Scientific, SA), and 0.1 mg/ml streptomycin (Thermo Fisher Scientific, SA).\u003c/p\u003e\u003cp\u003eTo achieve 97% confluency, the cells were harvested and counted using a hemocytometer. The Caco-2 cells were adjusted to a concentration of 1.5 \u0026times; 10\u003csup\u003e5\u003c/sup\u003e cells/ml by counting the cells that detached from the flask. The Caco-2 cells were subsequently seeded at 500 \u0026micro;l per well in a 36-well culture plate and incubated in a humidified 5% CO\u003csub\u003e2\u003c/sub\u003e incubator for 24 h at 37\u0026deg;C. The culture medium was changed every 2 days for 15 days to obtain a monolayer of differentiated Caco-2 cells.\u003c/p\u003e\u003cp\u003eThe adhesive activity of the isolates was examined using the method described by Fonseca et al. (\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e). \u003cem\u003eLactobacillus\u003c/em\u003e isolates were cultured in MRS broth at 37\u0026deg;C for 18 h. The grown bacterial cultures were then centrifuged at 8000 \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\times\\:\\)\u003c/span\u003e\u003c/span\u003e g for 10 min, and the cell pellets were collected, washed twice with sterile PBS, and then suspended in DMEM without antibiotics before being standardized to an OD600 nm of 1.0. Next, 1 ml of each bacterial suspension was added to the monolayer of differentiated Caco-2 cells in the wells of separate 36-well culture plates and incubated for 90 min at 37\u0026deg;C in a 5% CO\u003csub\u003e2\u003c/sub\u003e humidified incubator.\u003c/p\u003e\u003cp\u003eAfter incubation, the cells in the wells were washed three times with 1 ml of prewarmed PBS to remove any unbound \u003cem\u003eLactobacillus\u003c/em\u003e cells. Thereafter, the bounded Caco-2 cells and bacterial cells were detached from each well by adding 1 ml of Triton-X solution (0.1% v/v in PBS) and incubating for 10 min at 37\u0026deg;C. After incubation, the Caco-2 cells and \u003cem\u003eLactobacillus\u003c/em\u003e suspensions were harvested and centrifuged at 9000 \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\times\\:\\)\u003c/span\u003e\u003c/span\u003e g for 10 min at 4\u0026deg;C. The resulting pellet was washed twice with prewarmed PBS before being resuspended in PBS. Finally, 100 \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\mu\\:\\)\u003c/span\u003e\u003c/span\u003el of appropriate serial dilutions of the Caco-2 cell/bacterial cell suspension were spread on MRS agar plates and incubated at 37\u0026deg;C for 48 h before counting.\u003c/p\u003e\u003cp\u003eAdhesion ability (%) \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\frac{\\text{I}\\text{n}\\text{i}\\text{t}\\text{i}\\text{a}\\text{l}\\:\\text{c}\\text{o}\\text{u}\\text{n}\\text{t}\\text{s}\\:\\text{o}\\text{f}\\:\\text{b}\\text{a}\\text{c}\\text{t}\\text{e}\\text{r}\\text{i}\\text{a}\\text{l}\\:\\text{s}\\text{e}\\text{e}\\text{d}\\text{e}\\text{d}\\:}{\\text{T}\\text{h}\\text{e}\\:\\text{c}\\text{o}\\text{u}\\text{n}\\text{t}\\text{s}\\:\\text{a}\\text{f}\\text{t}\\text{e}\\text{r}\\:\\text{t}\\text{h}\\text{e}\\:\\text{w}\\text{a}\\text{s}\\text{h}\\text{i}\\text{n}\\text{g}\\:\\text{s}\\text{t}\\text{e}\\text{p}\\text{s}}\\)\u003c/span\u003e\u003c/span\u003e \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\times\\:\\)\u003c/span\u003e\u003c/span\u003e 100\u003c/p\u003e\u003cp\u003e\u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG was used as a positive control.\u003c/p\u003e\u003cp\u003e\u003cb\u003eCumulative probiotic potential (CPP) of\u003c/b\u003e \u003cb\u003eLactobacillus\u003c/b\u003e \u003cb\u003eisolates\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe probiotic potential of the 11 isolates from this study was assessed based on their cumulative probiotic score. The sum of acid tolerance, bile salt tolerance, auto-aggregation ability, cell surface hydrophobicity, antimicrobial activity, DNase activity, haemolytic activity, and antibiotic susceptibility is the CPP (\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eProbiotic Potential (%) = \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\frac{\\text{O}\\text{b}\\text{s}\\text{e}\\text{r}\\text{v}\\text{e}\\text{d}\\:\\text{s}\\text{c}\\text{o}\\text{r}\\text{e}}{\\text{M}\\text{a}\\text{x}\\text{i}\\text{m}\\text{u}\\text{m}\\:\\text{s}\\text{c}\\text{o}\\text{r}\\text{e}}\\)\u003c/span\u003e\u003c/span\u003e \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\times\\:\\)\u003c/span\u003e\u003c/span\u003e 100 (\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e)\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec15\" class=\"Section2\"\u003e\u003ch2\u003eStatistical analysis\u003c/h2\u003e\u003cp\u003eAll experiments were performed in triplicate, and descriptive statistics are presented as the mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation (SD). One-way analysis of variance (ANOVA) was employed, and Duncan\u0026rsquo;s multiple range test was used to compare significant differences between the mean values of acid tolerance, bile salt tolerance, auto-aggregation ability, cell surface hydrophobicity, cholesterol-lowering ability, antibiotic susceptibility, antioxidant activity, and cell adhesion. The Statistical Package for the Social Sciences (SPSS) program, version 29, was used for all analyses, and significance was considered at P\u0026thinsp;\u0026le;\u0026thinsp;0.05.\u003c/p\u003e\u003c/div\u003e"},{"header":"Results","content":"\u003cp\u003e\u003cstrong\u003eIdentification and phylogenetic analysis of \u003cem\u003eLactobacillus\u0026nbsp;\u003c/em\u003eisolates\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eLactobacillus\u003c/em\u003e species were identified in all the sources; a total of 18 presumptive \u003cem\u003eLactobacillus\u003c/em\u003e isolates were obtained from the samples and characterized as catalase-negative, non-motile, Gram-positive, and rod-shaped. These \u003cem\u003eLactobacillus\u003c/em\u003e isolates were identified as: \u003cem\u003eLactobacillus johnsonii\u0026nbsp;\u003c/em\u003e(n=7; 38.9%), \u003cem\u003eLatilactobacillus curvatus\u0026nbsp;\u003c/em\u003e(n=4; 22.2%), \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e (n=4; 22.2%), \u003cem\u003eLimosilactobacillus\u003c/em\u003e\u003cem\u003e\u0026nbsp;reuteri\u0026nbsp;\u003c/em\u003e(n=2; 11.1%), and \u003cem\u003eLatilactobacillus sakei\u003c/em\u003e (n=1; 5.6%) (\u003cstrong\u003eTable 1 \u0026amp; Table 2\u003c/strong\u003e).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eAs presented in \u003cstrong\u003eTable 1\u003c/strong\u003e,\u003cem\u003eLactobacillus\u003c/em\u003e \u003cem\u003ejohnsonii\u0026nbsp;\u003c/em\u003ewas the dominant\u003cem\u003e\u0026nbsp;Lactobacillus\u003c/em\u003e species isolated from the chicken gut, while \u003cem\u003eLactiplantibacillus\u003c/em\u003e\u003cem\u003e\u0026nbsp;plantarum\u0026nbsp;\u003c/em\u003ewas the prevalent species identified in vegetable samples. \u0026nbsp;On the other hand, \u003cem\u003eLatilactobacillus\u003c/em\u003e\u003cem\u003e\u0026nbsp;curvatus\u003c/em\u003e was the only\u003cem\u003e\u0026nbsp;Lactobacillus\u003c/em\u003e strain isolated from the banana samples.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 1\u003c/strong\u003e: \u003cem\u003eLactobacillus\u003c/em\u003e species identified from chicken gut, tomatoes, cucumber and banana\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 2\u003c/strong\u003e: Identification of \u003cem\u003eLactobacillus\u003c/em\u003e species isolated from chicken gut, tomato, cucumber, and banana using 16S rRNA Gene Sequencing\u003c/p\u003e\n\u003cp\u003eThe results depicted in \u003cstrong\u003eFigure 1\u0026nbsp;\u003c/strong\u003erevealed that the \u003cem\u003eLactobacillus\u0026nbsp;\u003c/em\u003estrainsare present in three distinct clades. One clade consisted of the \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e strains Chcg (chicken gut) and Pb2 (banana), establishing their close genetic relationship with \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e strains P1-S-OK326292 and 1224 MT573677 and \u003cem\u003eLatilactobacillus sakei\u0026nbsp;\u003c/em\u003estrain 6699 MT463883. These findings demonstrated the possible evolution of these isolates from a similar ancestor.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe second clade consisted of \u003cem\u003eLactiplantibacillus plantarum\u0026nbsp;\u003c/em\u003estrains To3a (tomato), To3d (tomato), and \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e strain Chcb (chicken gut), having a close relationship with \u003cem\u003eLatilactobacillus sakei\u0026nbsp;\u003c/em\u003estrain HBUAS5613 and \u003cem\u003eLatilactobacillus curvatus\u0026nbsp;\u003c/em\u003estrain TM332D OM265415, clearly indicating that they share a common ancestor.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eLactiplantibacillus plantarum\u0026nbsp;\u003c/em\u003estrain (To3d, tomato) clustered closely with \u003cem\u003eLactiplantibacillus plantarum\u0026nbsp;\u003c/em\u003estrain LP 212 OMO 38099, indicating a shared ancestry. In contrast, \u003cem\u003eLactiplantibacillus plantarum\u0026nbsp;\u003c/em\u003estrain (To3a, tomato) was differently clustered along \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e strain LP 212 OMO 38099, \u003cem\u003eLatilactobacillus sakei\u0026nbsp;\u003c/em\u003estrain HBUA S56133 MT211361 and \u003cem\u003eLatilactobacillus curvatus\u0026nbsp;\u003c/em\u003estrain TMPC 3332D OMO 265415, implying that they may not share a similar ancestor.\u003c/p\u003e\n\u003cp\u003eOn the other hand, \u003cem\u003eLactiplantibacillus plantarum\u0026nbsp;\u003c/em\u003estrains (To3b, tomatoes and Cu3e, cucumber), \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e strain (Cu2f, cucumber) and \u003cem\u003eLatilactobacillus sakei\u003c/em\u003e strain (Mcg, chicken gut) were clustered within the third clade. These strains formed a distinct sub-clade, indicating that they share a different common ancestor from the other isolates. \u003cem\u003eLatilactobacillus sakei\u003c/em\u003e strain (Mcg, chicken gut) shares a common ancestor with \u003cem\u003eLatilactobacillus sakei\u003c/em\u003e strains 2751 MT611760, 2178 MT604678 and SDCM 1005 MW548739.\u003c/p\u003e\n\u003cp\u003eAs shown in \u003cstrong\u003eFigure 2\u003c/strong\u003e, the phylogenetic tree indicates that \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e strain (Chcx4a, chicken gut) is closely related to \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e strain 20678 MW866875 in one clade but also connects to multiple branches of subclades consisting of \u003cem\u003eLactobacillus johnsonii\u0026nbsp;\u003c/em\u003estrains (Chcx2, Chcx3a, and Mcxb, chicken gut).\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eLactobacillus johnsonii\u0026nbsp;\u003c/em\u003estrains Mcyc, Mcxc and Mcya (chicken gut) were closely related to \u003cem\u003eLactobacillus johnsonii\u0026nbsp;\u003c/em\u003estrains MH33 FJ5422 and 33 PP728187, indicating a shared ancestry. In contrast, \u003cem\u003eLimosilolactobacillus reuteri\u0026nbsp;\u003c/em\u003estrains Chcx3a and Mcyc (chicken gut) exhibited a distant phylogenetic relationship with the other strains, suggesting that they do not share a common ancestor with the other strains.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcid tolerance\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe isolates \u003cem\u003eLactiplantibacillus plantarum\u0026nbsp;\u003c/em\u003e(To3b and To3d, tomato), \u003cem\u003eLimosilactobacillus\u003c/em\u003e\u003cem\u003e\u0026nbsp;reuteri\u0026nbsp;\u003c/em\u003e(Chx3a and Mcyc, chicken gut), \u003cem\u003eLactobacillus johnsonii\u0026nbsp;\u003c/em\u003e(Chx3b, Mcxb, and Mcxc, chicken gut), and \u003cem\u003eLatilactobacillus curvatus\u0026nbsp;\u003c/em\u003e(Cu2f, cucumber), displayed acid tolerance rates that were not significantly (p \u0026gt;0.05) different between pH 3.0 and pH 2.5. Whereas a lower survival rate significantly (p\u0026gt;0.05) different from the rest of the isolates was demonstrated by isolates \u003cem\u003eLatilactobacillus curvatus\u0026nbsp;\u003c/em\u003e(Cu2f, cucumber) and \u003cem\u003eLactobacillus johnsonii\u0026nbsp;\u003c/em\u003e(Mcxb and Mcxc, chicken gut) at pH 2.5 (\u003cstrong\u003eTable 3\u003c/strong\u003e).\u003c/p\u003e\n\u003cp\u003eThe isolates \u003cem\u003eLactiplantibacillus plantarum\u0026nbsp;\u003c/em\u003e(To3b and To3d, tomato), \u003cem\u003eLactiplantibacillus plantarum\u0026nbsp;\u003c/em\u003e(Cu3e, cucumber), \u003cem\u003eLatilactobacillus curvatus\u0026nbsp;\u003c/em\u003e(Pb2, banana), \u003cem\u003eLimosilactobacillus reuteri\u0026nbsp;\u003c/em\u003e(Mcyc, chicken gut), \u003cem\u003eLimosilactobacillus reuteri\u0026nbsp;\u003c/em\u003e(Chx3a, chicken gut), and \u003cem\u003eLacticaseibacillus rhamnosus\u0026nbsp;\u003c/em\u003eGGexhibited significantly (p \u0026le; 0.05) higher survival rates at pH 3, with no significant differences (p\u0026gt;0.05) observed among isolates \u003cem\u003eLactiplantibacillus plantarum\u0026nbsp;\u003c/em\u003e(To3a, tomato), \u003cem\u003eLimosilactobacillus reuteri\u0026nbsp;\u003c/em\u003e(Mcyc, chicken gut), and \u003cem\u003eLacticaseibacillus rhamnosus\u0026nbsp;\u003c/em\u003eGG. Meanwhile, isolates \u003cem\u003eLactilactobacillus curvatus\u0026nbsp;\u003c/em\u003e(Cu2f, cucumber) and\u003cem\u003e\u0026nbsp;Lactobacillus johnsonii\u0026nbsp;\u003c/em\u003e(Mcxc, chicken gut) exhibited relatively low survival rates of 53.79% and 65.65%, respectively, which were significantly different (p \u0026gt; 0.05) from the rates of the other isolates at pH 3(\u003cstrong\u003eTable 3\u003c/strong\u003e).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eOut of the 18 isolates analysed above, seven, \u003cem\u003eLatilactobacillus sakei\u0026nbsp;\u003c/em\u003e(Mcg, chicken gut), \u003cem\u003eLactobacillus johnsonii\u0026nbsp;\u003c/em\u003e(Chcx4a, Chcx2, Mcya, and Mcyb, chicken gut), \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e (Chcb and Chcg, chicken gut), exhibited a survival rate between 7% to 20% (data not shown) at pH 3.0 and hence were excluded from the probiotic due to their poor acidic tolerance, resulting in the total of 11\u003cem\u003e\u0026nbsp;Lactobacillus\u003c/em\u003e isolates used for further analysis.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 3:\u003c/strong\u003e The acid tolerance level of\u003cem\u003e\u0026nbsp;Lactobacillus\u003c/em\u003e isolates after exposure to pH 2.5 and pH 3.0 for 3h\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eBile salt tolerance\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe isolates \u003cem\u003eLactiplantibacillus plantarum\u0026nbsp;\u003c/em\u003e(To3b and To3d, tomato), \u003cem\u003eLimosilactobacillus\u003c/em\u003e\u003cem\u003e\u0026nbsp;reuteri\u0026nbsp;\u003c/em\u003e(Chx3a and Mcyc, chicken gut), \u003cem\u003eLactobacillus johnsonii\u0026nbsp;\u003c/em\u003e(Chx3b, Mcxb, and Mcxc, chicken gut), \u003cem\u003eLactiplantibacillus plantarum\u0026nbsp;\u003c/em\u003e(Cu3e, cucumber) and \u003cem\u003eLatilactobacillus curvatus\u0026nbsp;\u003c/em\u003e(Pb2, banana) exhibited bile salt tolerance that was not significantly (p \u0026gt; 0.05) different between 0.3% bile salt and 0.5% bile salt. On the other hand, the other isolates, including the reference strain, had a significantly (p \u0026le; 0.05) lower cell survival rate at 0.5% bile salt (\u003cstrong\u003eTable 4\u003c/strong\u003e).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 4:\u003c/strong\u003e The bile salt tolerance level of \u003cem\u003eLactobacillus\u003c/em\u003e isolates after exposure to 0.3% and 0.5% bile salt for 4 h\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eBSH activity\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThree isolates, \u003cem\u003eLactiplantibacillus plantarum\u0026nbsp;\u003c/em\u003e(To3a, tomato), \u003cem\u003eLimosilactobacillus\u003c/em\u003e\u003cem\u003e\u0026nbsp;reuteri\u0026nbsp;\u003c/em\u003e(Chcx3a, chicken gut) and \u003cem\u003eLactiplantibacillus plantarum\u0026nbsp;\u003c/em\u003e(Cu3e, cucumber) revealed the largest precipitation zones 10\u0026ndash;15 mm (++) compared with the reference strain and other isolates. However, \u003cem\u003eLactobacillus johnsonii\u0026nbsp;\u003c/em\u003e(Mcxc, chicken gut) showed no precipitation zone in this study (\u003cstrong\u003eTable 5\u003c/strong\u003e).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 5:\u0026nbsp;\u003c/strong\u003eThe BSH activity, EPS production ability, and cholesterol-lowering ability of \u003cem\u003eLactobacillus\u0026nbsp;\u003c/em\u003eisolates\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eThe EPS producing ability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eMost isolates, \u003cem\u003eLactiplantibacillus plantarum\u0026nbsp;\u003c/em\u003e(To3a, To3b, To3d, tomato), \u003cem\u003eLactiplantibacillus plantarum\u0026nbsp;\u003c/em\u003e(Cu3e, cucumber), \u003cem\u003eLimosilactobacillus\u003c/em\u003e\u003cem\u003e\u0026nbsp;reuteri\u0026nbsp;\u003c/em\u003e(Chx3a, chicken gut), \u003cem\u003eLatilactobacillus curvatus\u0026nbsp;\u003c/em\u003e(Cu2f, cucumber), and \u003cem\u003eLactobacillus johnsonii\u0026nbsp;\u003c/em\u003e(Mcxb, chicken gut), showed the ability to produce EPS, similar to the reference strain. Conversely, \u003cem\u003eLactobacillus johnsonii\u0026nbsp;\u003c/em\u003e(Chx3b and Mcxc, chicken gut), \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e (Pb2, banana), and \u003cem\u003eLimosilactobacillus\u003c/em\u003e\u003cem\u003e\u0026nbsp;reuteri\u0026nbsp;\u003c/em\u003e(Mcyc, chicken gut) did not exhibit the ability to produce EPS (\u003cstrong\u003eTable 5\u003c/strong\u003e).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCholesterol-lowering ability of \u003cem\u003eLactobacillus\u003c/em\u003e isolates\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe tested isolates exhibited varying degrees of cholesterol-reducing abilities, ranging from 13.06% to 57.64% (\u003cstrong\u003eTable 5\u003c/strong\u003e). The isolates \u003cem\u003eLactiplantibacillus plantarum\u0026nbsp;\u003c/em\u003e(To3a and To3d, tomato), \u003cem\u003eLimosilactobacillus\u003c/em\u003e\u003cem\u003e\u0026nbsp;reuteri\u0026nbsp;\u003c/em\u003e(Chx3a, chicken gut), and \u003cem\u003eLactiplantibacillus plantarum\u0026nbsp;\u003c/em\u003e(Cu3e, cucumber) exhibited significantly (p \u0026le;0.05) higher cholesterol-lowering abilities than the reference strain (41.81%). Isolates To3d and Chx3a exhibited a cholesterol-lowering ability of more than 50%. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDPPH scavenging activity\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe tested isolates exhibited varying degrees of DPPH scavenging activity, ranging from 6.81% to 24.45% compared with 35.63% from ascorbic acid. The DPPH scavenging activity of \u003cem\u003eLactiplantibacillus plantarum\u0026nbsp;\u003c/em\u003e(To3d, tomato) (24.45%) was significantly (p \u0026le; 0.05) higher than that of the reference strain (23.20%). In contrast, that of \u003cem\u003eLimosilactobacillus\u003c/em\u003e\u003cem\u003e\u0026nbsp;reuteri\u0026nbsp;\u003c/em\u003e(Mcyc,\u0026nbsp;chicken\u0026nbsp;gut)\u0026nbsp;(20.90%) was not significantly different (P\u0026gt;0.05) from that of the reference strain (\u003cstrong\u003eTable 6\u003c/strong\u003e).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 6:\u003c/strong\u003e DPPH scavenging activity (%) of \u003cem\u003eLactobacillus\u003c/em\u003e isolates\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eA\u003c/strong\u003e\u003cstrong\u003entimicrobial activity against pathogenic bacteria\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe tested \u003cem\u003eLactobacillus\u003c/em\u003e isolates exhibited varying degrees of antimicrobial activity against selected pathogens. The isolates \u003cem\u003eLactiplantibacillus plantarum\u0026nbsp;\u003c/em\u003e(To3a, To3b, and To3d, tomato) demonstrated antimicrobial activity similar to that of the reference strain, with an inhibition zone of 10 to 15 mm (++) or more against \u003cem\u003eE. coli\u003c/em\u003e, \u003cem\u003eS. aureus\u003c/em\u003e, \u003cem\u003eB. cereus\u003c/em\u003e, and \u003cem\u003eL. monocytogenes\u0026nbsp;\u003c/em\u003e(\u003cstrong\u003eTable 7\u003c/strong\u003e).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 7:\u003c/strong\u003e Antimicrobial activity of CFS of \u003cem\u003eLactobacillus\u003c/em\u003e against pathogenic organisms\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCell auto-aggregation\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe auto-aggregation (%) of selected \u003cem\u003eLactobacillus\u003c/em\u003e isolates generally increased after incubation from 2 to 4 h, as shown in \u003cstrong\u003eTable 8\u003c/strong\u003e. Compared with the reference strain, the isolates \u003cem\u003eLactiplantibacillus plantarum\u0026nbsp;\u003c/em\u003e(To3a, To3b and To3d, tomato), \u003cem\u003eLimosilactobacillus\u003c/em\u003e\u003cem\u003e\u0026nbsp;reuteri\u0026nbsp;\u003c/em\u003e(Chx3a and Mcyc, chicken gut), \u003cem\u003eLactobacillus johnsonii\u0026nbsp;\u003c/em\u003e(Chx3b, chicken gut), and \u003cem\u003eLatilactobacillus curvatus\u0026nbsp;\u003c/em\u003e(Cu2f, cucumber),\u0026nbsp;displayed a significant\u0026nbsp;(p \u0026le; 0.05)\u0026nbsp;increase in auto-aggregation (%) after incubation from 2 to 4 h. Similarly, the auto-aggregation (%) of these isolates was significantly\u0026nbsp;(p \u0026le; 0.05)\u0026nbsp;higher than that of the reference strain after incubation for 2 and 4 h, respectively. The other strains displayed auto-aggregation (%) between 18.00% and 30.67% after 4 h incubation, which was significantly different (p\u0026le; 0.05) from that\u0026nbsp;demonstrated by the reference strain at 24.67%.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 8:\u0026nbsp;\u003c/strong\u003eTheauto-aggregation ability of \u003cem\u003eLactobacillus\u003c/em\u003e isolates after 2 h and 4 h incubation\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCell surface hydrophobicity\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe \u003cem\u003eLactobacillus\u003c/em\u003e isolates exhibited varying degrees of cell surface hydrophobicity (\u003cstrong\u003eTable 9\u003c/strong\u003e). The \u003cem\u003eLatilactobacillus curvatus\u0026nbsp;\u003c/em\u003e(Cu2f, cucumber), \u003cem\u003eLactiplantibacillus plantarum\u0026nbsp;\u003c/em\u003e(Cu3e, cucumber), and \u003cem\u003eLimosilactobacillus\u003c/em\u003e\u003cem\u003e\u0026nbsp;reuteri\u0026nbsp;\u003c/em\u003e(Mcyc, chicken gut) showed cell surface hydrophobicity values significantly (p \u0026le; 0.05) higher than those of \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG,whereas \u003cem\u003eLactobacillus johnsonii\u0026nbsp;\u003c/em\u003e(Chx3b, chicken gut)\u0026nbsp;exhibited cell surface hydrophobicity (%) not significantly different (p\u0026gt;0.05) from those of the reference strain. The remaining isolates\u0026nbsp;presented a\u0026nbsp;cell surface hydrophobicity\u0026nbsp;(%) between 0.00% and 46.67% after 1 h of incubation, which was significantly different (p \u0026le; 0.05) from that of the reference strain (53.33%).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 9:\u0026nbsp;\u003c/strong\u003eCell surface hydrophobicity ability of \u003cem\u003eLactobacillus\u003c/em\u003e isolates after 1 hr incubation\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAntibiotic susceptibility\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe \u003cem\u003eLactobacillus\u003c/em\u003e isolates showed varying antibiotic susceptibility profiles. All the isolates were susceptible to penicillin G (10 \u0026mu;g), chloramphenicol (30 \u0026mu;g), erythromycin (15 \u0026mu;g) and ampicillin (10 \u0026mu;g). On the contrary, all the isolates were resistant to kanamycin (30 \u0026mu;g), gentamycin (10 \u0026mu;g), and streptomycin (10 \u0026mu;g) (\u003cstrong\u003eTable 10\u003c/strong\u003e).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 10:\u003c/strong\u003e Antibiotic susceptibility profile of \u003cem\u003eLactobacillus\u003c/em\u003e isolates using the disk diffusion method\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eHaemolysis and DNase activity\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNone of the \u003cem\u003eLactobacillus\u003c/em\u003e isolates in this study exhibited haemolysis or DNase activity (\u003cstrong\u003eTable 11\u003c/strong\u003e).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 11:\u0026nbsp;\u003c/strong\u003eBloodhaemolysis and DNase activity of \u003cem\u003eLactobacillus\u003c/em\u003e isolates\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCell adhesion assay\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eLactobacillus johnsonii\u0026nbsp;\u003c/em\u003e(Chx3b, chicken gut) and \u003cem\u003eLactiplantibacillus plantarum\u0026nbsp;\u003c/em\u003e(To3d, tomato) showed Caco-2 cell adhesion capacities (15.29% and 12.87% respectively) higher than that of the reference strain (11.40%), whereas the isolate \u003cem\u003eLimosilactobacillus reuteri\u0026nbsp;\u003c/em\u003e(Mcyc, chicken gut) presented a slightly lower adhesion capacity of 10.53%. In contrast, isolates \u003cem\u003eLatilactobacillus curvatus\u0026nbsp;\u003c/em\u003e(Pb2, banana) and \u003cem\u003eLactiplantibacillus plantarum\u0026nbsp;\u003c/em\u003e(Cu3e, cucumber)\u0026nbsp;demonstrated the lowest adhesion capacities at 2.60% and 3.58%, respectively (\u003cstrong\u003eTable 12\u003c/strong\u003e).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 12:\u0026nbsp;\u003c/strong\u003eAdhesion capacity of \u003cem\u003eLactobacillus\u003c/em\u003e isolates to Caco-2 epithelial cells\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCPP of \u003cem\u003eLactobacillus\u003c/em\u003e isolates\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe CPP of the eleven isolates, as presented in \u003cstrong\u003eTable 13\u003c/strong\u003e, are 50% for isolate \u003cem\u003eLatilactobacillus curvatus\u0026nbsp;\u003c/em\u003e(Pb2, banana), and \u003cem\u003eLactobacillus johnsonii\u0026nbsp;\u003c/em\u003e(Mcxb, chicken gut); 62.50% for isolate \u003cem\u003eLimosilactobacillus reuteri\u0026nbsp;\u003c/em\u003e(Chx3a, chicken gut), \u003cem\u003eLactiplantibacillus plantarum\u0026nbsp;\u003c/em\u003e(Cu3e, cucumber), and \u003cem\u003eLactobacillus johnsonii\u0026nbsp;\u003c/em\u003e(Mcxc, chicken gut); 75% for isolate \u003cem\u003eLactiplantibacillus plantarum\u0026nbsp;\u003c/em\u003e(To3b, tomato), \u003cem\u003eLactobacillus johnsonii\u0026nbsp;\u003c/em\u003e(Chx3b, chicken gut), and \u003cem\u003eLimosilactobacillus reuteri\u0026nbsp;\u003c/em\u003e(Mcyc, chicken gut), and 87.50% for the remaining three isolates: \u003cem\u003eLactiplantibacillus plantarum\u0026nbsp;\u003c/em\u003e(To3a and To3d, tomato) \u0026nbsp;and \u003cem\u003eLatilactobacillus curvatus\u0026nbsp;\u003c/em\u003e(Cu2f, cucumber).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 13:\u0026nbsp;\u003c/strong\u003eCumulative probiotic potential of \u003cem\u003eLactobacillus\u003c/em\u003e isolates\u003c/p\u003e"},{"header":"Discussions","content":"\u003cp\u003e\u003cb\u003eIdentification of\u003c/b\u003e \u003cb\u003eLactobacillus\u003c/b\u003e \u003cb\u003estrains\u003c/b\u003e\u003c/p\u003e\u003cp\u003e\u003cem\u003eLactobacillus\u003c/em\u003e species were identified in all the sample types, possibly because they are widely distributed in various ecological niches and are adapted to survive naturally in nutrient-rich and moist environments, which aids their growth and colonization (\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eAmong several species of \u003cem\u003eLactobacillus\u003c/em\u003e strains, \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e and \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e have been observed to be among the most detected microbes in poultry GIT (\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e). Additionally, Junnarkar et al. (\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e) reported in their study that the genus \u003cem\u003eLactobacillus\u003c/em\u003e was dominant in fresh vegetable samples (cabbage, cauliflower, cluster bean, French beans, gherkins, bitter gourd, rigid gourd, fenugreek, and tomatoes). In particular, \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e was one of the most prevalent bacterial species found in tomatoes, fermented cabbage, and olives (\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eBacterial strains derived from plant-based matrices, such as fruits and vegetables, and the animal gut, possess notable genetic and functional diversity that the food industry may leverage to foster innovative probiotic applications (\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cem\u003eLactobacillus johnsonii\u003c/em\u003e is a resident microflora of the poultry GIT that particularly adheres to the epithelium of the duodenum and jejunum (\u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e, \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cem\u003eLactobacillus johnsonii\u003c/em\u003e was the most prevalent \u003cem\u003eLactobacillus\u003c/em\u003e species in the chicken gut samples, possibly because of external factors, such as chicken feed, antibiotic treatment, and invading organisms (\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e). Furthermore, the level of stress, environmental conditions, and animal health may also play a role in the differences in the microbial composition of the chicken gut (\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eSimilar to our findings, a relatively high occurrence of \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e (strains AER105 and AER25) isolated from the ilea of five individual chickens (\u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e), and \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e; 21% in chicken mucosa of the ileum (M-IM), 15% in the cecal lumen (M-CL) and 11% in the mucosa of ceca (M-CM) (\u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e53\u003c/span\u003e) was established in their study.\u003c/p\u003e\u003cp\u003eHowever, previous studies have reported that \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e isolated from the GIT of chickens is prevalent in 23.3% (\u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e); 10% (\u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e55\u003c/span\u003e), 7.69% (\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e), and 5% (\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e(\u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e56\u003c/span\u003e) mentioned that the type of chicken breed can influence their gut microbiota, for example, laying hens have different microbial communities than the broiler breeds used in this study. This is owing to their physiological and genetic variations (\u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e57\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eIn addition, feed formulations, including the use of probiotics, protein, and fibre in the chicken diet, can favourably impact the overall microbial population (\u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e56\u003c/span\u003e, \u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e58\u003c/span\u003e). According to Bindari \u0026amp; Gerber (\u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e59\u003c/span\u003e), higher levels of \u003cem\u003eLactobacillus\u003c/em\u003e species were noted throughout the GIT of chickens fed whole wheat than in chickens fed grounded wheat pellets.\u003c/p\u003e\u003cp\u003eHousing systems, such as the use of cages, barns and free-range systems (\u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e57\u003c/span\u003e), and the geographical location of the chickens (high altitude or low altitude regions) may also reshape the microbiota landscape and colonization (\u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e56\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe prevalence of \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e represents an important tool that could be utilized as probiotics in poultry feed for improving the gut microbial structure, weight gain, enhancing feed conversion ratio, and overall performance, especially in boilers (\u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e60\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe next most dominant microbe was \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e, which was isolated from the chicken gut, banana and cucumber.\u003c/p\u003e\u003cp\u003e\u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e is commonly associated with lactic acid fermentation and can be isolated from several environments, including fermented meat, fermented vegetables, dairy products, human GIT, and other niches (\u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e61\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e is mostly associated with fermented meat and poultry products (\u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e62\u003c/span\u003e), although it may be isolated from foods of vegetable origin as well as the GIT of animals that feed on plants or cereals, including chickens (\u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e63\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e can inhibit the growth of foodborne pathogens in fermented food products via the production of bacteriocin (\u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e61\u003c/span\u003e). Besides, they play an essential role in the preservation and flavour development of fermented dairy foods via lactic acid production and the development of desirable flavours in meat products (\u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e64\u003c/span\u003e). Additionally, the incorporation of \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e strains such as SMFM2016-NK in fermented milk may lessen periodontitis and regulate oral and gut microbial composition (\u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e65\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e sourced from tomatoes and cucumbers was the dominant strain in this study. This microbe is predominantly found in fermented food, including cucumbers and tomatoes (\u003cspan citationid=\"CR66\" class=\"CitationRef\"\u003e66\u003c/span\u003e, \u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e67\u003c/span\u003e), and plant surfaces (\u003cspan citationid=\"CR68\" class=\"CitationRef\"\u003e68\u003c/span\u003e), and it is one of the most versatile and metabolically diverse organisms commonly involved in vegetable fermentation of products such as kefir and sauerkraut (\u003cspan citationid=\"CR69\" class=\"CitationRef\"\u003e69\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eA previous study revealed that they can enhance their genome for growth in an environment rich in plant carbohydrates, owing to the incidence of genomic islands sheltering mosaic modules or cassettes of carbohydrate utilization genes, which may have been acquired through horizontal gene transfer (\u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e70\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eBorjihan et al. (\u003cspan citationid=\"CR71\" class=\"CitationRef\"\u003e71\u003c/span\u003e) mentioned that \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e has a distinct adaptability to the vegetable environment, which may be attributed to its stress response pathways and large metabolic capacity. Additionally, the ability of this bacterial species to produce antimicrobial compounds and biofilm capacity are attributes that enable it to survive and dominate in such an environment (\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e was reported to be prevalent in 56.82% of raw tomatoes (\u003cspan citationid=\"CR72\" class=\"CitationRef\"\u003e72\u003c/span\u003e), 34.6% of vegetable residues and silage fermentation (\u003cspan citationid=\"CR73\" class=\"CitationRef\"\u003e73\u003c/span\u003e), 55.55% (\u003cspan citationid=\"CR74\" class=\"CitationRef\"\u003e74\u003c/span\u003e) and 57.69% in Chinese fermented vegetables (\u003cspan citationid=\"CR75\" class=\"CitationRef\"\u003e75\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eOn the contrary, 19.2% \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e was dominant in plant-based samples (\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e), 11.11% in fresh vegetables (\u003cspan citationid=\"CR76\" class=\"CitationRef\"\u003e76\u003c/span\u003e), and 3.89% in cabbage and tomatoes (\u003cspan citationid=\"CR77\" class=\"CitationRef\"\u003e77\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe probiotic \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e strain plays a pivotal role in the gut microbial regulation and reduction in total body fat and body weight gain, as demonstrated in a high-fat-diet obese mice trial (\u003cspan citationid=\"CR78\" class=\"CitationRef\"\u003e78\u003c/span\u003e). In addition, this bacterium may be a promising probiotic candidate for improving fermentation-based food products (\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eOn the other hand, \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e was the only \u003cem\u003eLactobacillus\u003c/em\u003e species isolated from the banana fruit samples. This fermentation bacterial strain is usually found in the fruit matrix and has remarkable fermentation properties, which could result in improved preservation and nutritional qualities (\u003cspan citationid=\"CR79\" class=\"CitationRef\"\u003e79\u003c/span\u003e). The reason for this may be that \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e bacteria possess numerous genes that encode metabolic pathways for carbohydrate utilization, such as starch, which acts as an ideal substrate that encourages the colonization and growth of this microbe (\u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e62\u003c/span\u003e) in bananas.\u003c/p\u003e\u003cp\u003eConversely, 35% \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e isolates were derived from Italian red and yellow pepper fruits (\u003cspan citationid=\"CR80\" class=\"CitationRef\"\u003e80\u003c/span\u003e), 38.19% from Saudi chicken ceca samples (\u003cspan citationid=\"CR81\" class=\"CitationRef\"\u003e81\u003c/span\u003e), and 20% from fermented Chinese sausage (\u003cspan citationid=\"CR82\" class=\"CitationRef\"\u003e82\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eSoil health and quality, for example, the planting of banana with high-fertility soil, may contribute to a robust bacterial community composition (\u003cspan citationid=\"CR83\" class=\"CitationRef\"\u003e83\u003c/span\u003e). In addition, Kaushal et al. (\u003cspan citationid=\"CR84\" class=\"CitationRef\"\u003e84\u003c/span\u003e) mentioned that differences in climatic conditions, such as temperature, rainfall, and geographical characteristics of banana-producing areas, may significantly contribute to variations in the microbial diversity of bananas.\u003c/p\u003e\u003cp\u003e\u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e isolate derived from bananas may hold a significant promise in the health industry due to its potential probiotic impact in modulating immune response, reducing colitis and improving gut barrier functions (\u003cspan citationid=\"CR85\" class=\"CitationRef\"\u003e85\u003c/span\u003e). In the food industry, they may serve as a bio-preservative agent by producing lactic acid, which is useful for the preservation and flavour development of fermented dairy food (\u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e64\u003c/span\u003e).\u003c/p\u003e\u003cdiv id=\"Sec29\" class=\"Section2\"\u003e\u003ch2\u003eAcid tolerance\u003c/h2\u003e\u003cp\u003eThe acid tolerance of selected \u003cem\u003eLactobacillus\u003c/em\u003e isolates generally increased at pH 3.0, as shown in Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e. The acidic pH of the stomach (2.5\u0026ndash;3.5) results in the formation of a natural barrier against external bacteria (\u003cspan citationid=\"CR86\" class=\"CitationRef\"\u003e86\u003c/span\u003e), requiring \u003cem\u003eLactobacillus\u003c/em\u003e species to tolerate such low pH conditions to survive and remain active during GIT transit, particularly over the estimated 3 h period it spends in the stomach during food passage (\u003cspan citationid=\"CR87\" class=\"CitationRef\"\u003e87\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe isolates, \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e (To3a, To3d, tomatoes), \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e (Cu3e, cucumber), \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e (Pb2, banana), \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e (Mcyc, Chx3a, chicken gut), and \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG, exhibited significantly (p\u0026thinsp;\u0026le;\u0026thinsp;0.05) higher survival rates at pH 3. Our results highlighted that these \u003cem\u003eLactobacillus\u003c/em\u003e species are highly acid-tolerant strains, and that their ability to survive in acidic medium differs among different species and sources.\u003c/p\u003e\u003cp\u003eMost \u003cem\u003eLactobacillus\u003c/em\u003e species can tolerate acidic environments (pH 2\u0026ndash;3) because of their remarkable ability to metabolize sugars to generate lactic acid (\u003cspan citationid=\"CR88\" class=\"CitationRef\"\u003e88\u003c/span\u003e). Furthermore, Hojjati et al. (\u003cspan citationid=\"CR89\" class=\"CitationRef\"\u003e89\u003c/span\u003e) and Barzegar et al. (\u003cspan citationid=\"CR90\" class=\"CitationRef\"\u003e90\u003c/span\u003e) reported that \u003cem\u003eLactobacillus\u003c/em\u003e can develop distinct mechanisms for survival in low acidic conditions, the most important of which is F\u003csub\u003e0\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e-ATPase. This mechanism shields the bacterial cell from acidic exposure by transferring the protons out of the cell membrane and thus maintaining the intracellular pH (\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e). Other mechanisms by which \u003cem\u003eLactobacillus\u003c/em\u003e species tolerate high acidity include H\u003csup\u003e+\u003c/sup\u003e pumping to an acidic environment, acid end-product efflux, the synthesis of alkali products to neutralize acid produced during extracellular metabolism, the repair of macromolecules, biofilm formation, and pre-adaptation and cross-protection (\u003cspan citationid=\"CR91\" class=\"CitationRef\"\u003e91\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eAdditionally, the EPS produced by \u003cem\u003eLactobacillus\u003c/em\u003e forms a layer in a low pH environment that limits the access of exogenous acids to the bacterial cells due to anions bound to EPS as a phosphate group (\u003cspan citationid=\"CR92\" class=\"CitationRef\"\u003e92\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eIn a separate study, \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e strains O19, O20, O21, O23, and K1 and \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e strain K4, which were isolated from traditional fermented cabbage and cucumber, also showed a high tolerance rate\u0026thinsp;\u0026gt;\u0026thinsp;90% to pH 2.5 and pH 3.5 (\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e). Ahire et al. (\u003cspan citationid=\"CR93\" class=\"CitationRef\"\u003e93\u003c/span\u003e) found that \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e UBLP40 isolated from indigenous fermented food had a 93% survival rate at pH 3.0 after 3 h of incubation.\u003c/p\u003e\u003cp\u003e\u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e strains sourced from Italian food products showed high tolerance to pH 3.0 and 2.5, presenting a survival rate ranging from 60.69% to 100.08% (\u003cspan citationid=\"CR94\" class=\"CitationRef\"\u003e94\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eIn another study, Rajoka et al. (\u003cspan citationid=\"CR95\" class=\"CitationRef\"\u003e95\u003c/span\u003e) reported that six \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e strains and one \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e strain sourced from the chicken GIT demonstrated a high survival rate greater than 80% at pH 3.0 after 3h of incubation, whereas five out of seven \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e strains showed a survival rate\u0026thinsp;\u0026gt;\u0026thinsp;70% at pH 3.0 after 5 h incubation (\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eHigh resistance to pH 3.0 conditions was also observed for 25 \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e strains, with bacterial growth rates ranging from 60.27% to 100.48% after 24 h of exposure (\u003cspan citationid=\"CR96\" class=\"CitationRef\"\u003e96\u003c/span\u003e). Meanwhile, the survival ability of \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e strains (AF(PIF), P8014, M8, LFS7 and a7) isolated from different sources at pH 2.5 was reported to be 98.85% to 100% after 4 h of exposure (\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e), while Tokatl et al.(\u003cspan citationid=\"CR95\" class=\"CitationRef\"\u003e95\u003c/span\u003e) reported that at pH 2.5, twenty-one \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e strains sourced from traditional pickles showed survival ability of 35% \u0026ndash; 85% after 4 h of exposure.\u003c/p\u003e\u003cp\u003eThese results highlight the potential role of these strains in the development of probiotic supplements without a coating (\u003cspan citationid=\"CR97\" class=\"CitationRef\"\u003e97\u003c/span\u003e), and in maintaining cell viability, especially in acidic food systems (\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e) like fermented fruits and vegetables.\u003c/p\u003e\u003cp\u003eThe survival rates of the other isolates did not differ significantly at pH 2.5 and pH 3. This is possibly due to the strain-specific nature of these strains and their remarkable resilience in an acidic environment (\u003cspan citationid=\"CR93\" class=\"CitationRef\"\u003e93\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe differences observed in our results regarding the acidic tolerance rates of lactobacilli isolates may be attributed to the bacterial source and the duration of acidic exposure (\u003cspan citationid=\"CR93\" class=\"CitationRef\"\u003e93\u003c/span\u003e). Furthermore, acid tolerance variation in LAB has been connected to the dissimilarity in H\u003csup\u003e+\u003c/sup\u003e-ATPase activity induction, leading to the removal of protons (H\u003csup\u003e+\u003c/sup\u003e), alkalinization of the outside environment, and alterations in the cell envelope composition (\u003cspan citationid=\"CR98\" class=\"CitationRef\"\u003e98\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eOur findings corroborate the report by Zielińska et al. (\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e), who found that at pH 2.5 and pH 3.5, there were no significant changes in the survival rates of \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e (O19, O20, O21, O23, and K1) and \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e (K4) isolates in their study, as all exhibited a survival rate greater than 90%. Manzoor \u0026amp; Tayyeb(\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e) revealed that all \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e isolates maintained a constant viable cell count at pH 3 after 4 h of exposure. In addition, \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e SW03 and SW07 were not significantly different, and their survival rate was above 90% when exposed to pH 2.5 and 3.0 after 3hr incubation (\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e). Likewise, no significant differences were recorded between \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e TF-7 and \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG after exposure to pH 2.0, 3.0, and 4.0 (\u003cspan citationid=\"CR99\" class=\"CitationRef\"\u003e99\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eIn contrast, an Italian study conducted by Turchi et al. (\u003cspan citationid=\"CR94\" class=\"CitationRef\"\u003e94\u003c/span\u003e) reported high growth rates, ranging from 96.98% to 102.87% at pH 3.0, were observed by all 37 \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e strains, while at pH 2.5, survival rates ranging from 60.69% to 100.08% were recorded.\u003c/p\u003e\u003cp\u003eFurthermore, Mulaw et al. (\u003cspan citationid=\"CR100\" class=\"CitationRef\"\u003e100\u003c/span\u003e) reported that \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e EO52 and TO35 were significantly different at pH 2.5 and 3.0, after exposure for 6 h.\u003c/p\u003e\u003cp\u003eThe isolates \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e (To3a, tomato) and \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e (Mcyc, chicken gut) exhibited survival rates of 97% and 98.1%, respectively, at pH 3, which were higher than those of \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG, with a survival rate of 93%.\u003c/p\u003e\u003cp\u003e\u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e and various homofermentative lactobacilli can balance their intracellular redox state using an extracellular electron transfer mechanism (\u003cspan citationid=\"CR101\" class=\"CitationRef\"\u003e101\u003c/span\u003e, \u003cspan citationid=\"CR102\" class=\"CitationRef\"\u003e102\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cem\u003eLactobacillus\u003c/em\u003e species of vegetable origin, such as tomato, are generally more pH-tolerant (\u003cspan citationid=\"CR75\" class=\"CitationRef\"\u003e75\u003c/span\u003e). Zhang et al. (\u003cspan citationid=\"CR103\" class=\"CitationRef\"\u003e103\u003c/span\u003e) pointed out that the resilience of \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e to low acidic concentration of pH 3.0 was positively linked with the production of EPS, as the low pH tolerance of these bacteria can be improved by adjusting their plasma membrane fatty acid composition to decrease fluidity, increasing the proton pump activity to maintain the intracellular pH balance. According to Nguyen et al. (\u003cspan citationid=\"CR92\" class=\"CitationRef\"\u003e92\u003c/span\u003e), an increase in biofilm formation in EPS-producing \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e strains was noted, compared with a decrease in the formation of biofilm in EPS-producing \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG under low pH conditions.\u003c/p\u003e\u003cp\u003eAdditionally, species like \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e naturally inhabit the human or animal small intestine; hence, they are better genetically adapted to the low pH conditions of the gut than those of food origin (\u003cspan citationid=\"CR104\" class=\"CitationRef\"\u003e104\u003c/span\u003e, \u003cspan citationid=\"CR105\" class=\"CitationRef\"\u003e105\u003c/span\u003e). Owing to their enhanced host adaptation, greater cell viability, and safety, probiotic strains isolated from their natural hosts, such as the animal gut, are chosen over those derived from other sources (\u003cspan citationid=\"CR106\" class=\"CitationRef\"\u003e106\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eOur results agree with those obtained from an earlier study, where \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e DUR5 and DUR8 demonstrated a higher survival rate of 87.61% and 90.24%, respectively, whereas \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG exhibited 80.61% resistance to pH 3.0 (\u003cspan citationid=\"CR107\" class=\"CitationRef\"\u003e107\u003c/span\u003e). A study by Jomehzadeh et al. (\u003cspan citationid=\"CR108\" class=\"CitationRef\"\u003e108\u003c/span\u003e) noted that \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e N12 to N20 showed survival rates ranging from 60% to 96% compared with 58% demonstrated by \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG at pH 2.5. In addition, \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e LpJ1 and LpJ5 demonstrated significantly higher viable cell counts of 7.93 CFU/ml and 7.93 CFU/ml, respectively, than did \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG (6.00 CFU/ml) at pH 2.5 after 180 min incubation (\u003cspan citationid=\"CR109\" class=\"CitationRef\"\u003e109\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eIn contrast, a higher survival rate of 86.48% was demonstrated by \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG exposed to gastric juice at pH 2.5 for 3 h, compared with \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e LN-3-1, which had a 38.06% survival rate (\u003cspan citationid=\"CR110\" class=\"CitationRef\"\u003e110\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e strain C06 exhibited tolerance levels greater than 90% when exposed to acidic conditions at pH 3 (\u003cspan citationid=\"CR111\" class=\"CitationRef\"\u003e111\u003c/span\u003e), but this value is less than the 110% survival rate achieved by the \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e H11 strain after 3h exposure to pH 3 (\u003cspan citationid=\"CR106\" class=\"CitationRef\"\u003e106\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eHowever, \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e L-3 displayed a moderate survival rate above 60% at pH 3.0, as observed by Wang et al. (\u003cspan citationid=\"CR112\" class=\"CitationRef\"\u003e112\u003c/span\u003e), and \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e strains reported viability of 58% \u0026ndash; 79% after 3 h exposure to pH 3 (\u003cspan citationid=\"CR113\" class=\"CitationRef\"\u003e113\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe disparity in \u003cem\u003eLactobacillus\u003c/em\u003e isolate resistance to low pH may be due to the diverse low pH tolerance traits of several lactobacilli species sourced from different habitats and geographical regions (\u003cspan citationid=\"CR114\" class=\"CitationRef\"\u003e114\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eHighly acid-tolerant strains, such as \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e and \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e, are important for modulating the oral and gut microbiota, promoting immune responses and reducing inflammation (\u003cspan citationid=\"CR103\" class=\"CitationRef\"\u003e103\u003c/span\u003e, \u003cspan citationid=\"CR105\" class=\"CitationRef\"\u003e105\u003c/span\u003e). Additionally, the \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e Mcyc strain isolated from this study may be crucial for developing a probiotic supplement for broiler chicken growth and development, while the \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e To3a strain could be a promising candidate for creating a probiotic plant-based product, such as tomato juice, since they are host-adapted (\u003cspan citationid=\"CR115\" class=\"CitationRef\"\u003e115\u003c/span\u003e) .\u003c/p\u003e\u003cp\u003eAmong the 18 isolates obtained from the different samples, seven isolates, namely, \u003cem\u003eLatilactobacillus sakei\u003c/em\u003e (Mcg, chicken gut), \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e (Chxc4 and Chcx2, chicken gut), \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e (Mcya and Mcyb, chicken gut), \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e (Chcg, and Chcb, chicken gut), exhibited a survival rate ranging from 7% \u0026ndash; 20% at pH 3.0, were excluded from the probiotic assay due to their poor tolerance to low pH, as HCl severely reduces the viable bacteria cell population in an acidic media (\u003cspan citationid=\"CR116\" class=\"CitationRef\"\u003e116\u003c/span\u003e). Khushboo et al.(\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e) highlighted that for an organism to be considered probiotic, it must be able to resist the low acidic conditions (pH 1.0\u0026ndash;3.0) necessary for survival in the host GIT. For this reason, isolates must display 50% tolerance to pH 3.0 for them to be considered resistant to low pH (\u003cspan citationid=\"CR87\" class=\"CitationRef\"\u003e87\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eSome bacterial cells lack a resistance mechanism when exposed to a stressful environment, such as a pH of 3.0, as proton influx at low pH disrupts the pH homeostasis mechanisms and reduces their intracellular pH (\u003cspan citationid=\"CR117\" class=\"CitationRef\"\u003e117\u003c/span\u003e). Cizeikiene \u0026amp; Jagelaviciute (\u003cspan citationid=\"CR118\" class=\"CitationRef\"\u003e118\u003c/span\u003e) mentioned that H\u003csup\u003e+\u003c/sup\u003e-ATPase activity is reduced for non-acid-tolerant strains when exposed to acidic conditions, consequently affecting the maintenance of an intracellular pH (\u003cspan citationid=\"CR118\" class=\"CitationRef\"\u003e118\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eAt pH 3.5, a 0% survival rate was detected for \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e UBLJ01 (vagina of healthy women) after 24 h of incubation via the traditional plate count method (\u003cspan citationid=\"CR119\" class=\"CitationRef\"\u003e119\u003c/span\u003e). In another study, the \u003cem\u003eLactobacillus\u003c/em\u003e isolates EKU 1 and EMB5, which were sourced from Ethiopian cottage cheese, showed 5% and 41.48% survival rates, respectively, at pH 3 after a 3 h incubation period (\u003cspan citationid=\"CR120\" class=\"CitationRef\"\u003e120\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThese poor acid-resistant isolates may be unable to grow and colonize in the stomach when utilized as probiotics; hence, they have low probiotic potential (\u003cspan citationid=\"CR121\" class=\"CitationRef\"\u003e121\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e\n\u003ch3\u003eBile tolerance\u003c/h3\u003e\n\u003cp\u003eThe human bile concentrations range from 0.3% to 0.5% (\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e, \u003cspan citationid=\"CR109\" class=\"CitationRef\"\u003e109\u003c/span\u003e), hence, any candidate probiotic bacteria must be able to survive the above-mentioned bile concentration (\u003cspan citationid=\"CR69\" class=\"CitationRef\"\u003e69\u003c/span\u003e), considering that bile salts are synthesized in the liver and disrupt the cell membrane, thus resulting in DNA damage and oxidative stress (\u003cspan citationid=\"CR122\" class=\"CitationRef\"\u003e122\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe isolates \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e (To3b and To3d, tomato), \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e (Mcyc, chicken gut), \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e (Chx3b, Mcxb, and Mcxc, chicken gut), \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e (Cu3e, cucumber) and \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e (Pb2, banana) exhibited bile salt tolerances that were not significantly (p\u0026thinsp;\u0026gt;\u0026thinsp;0.05) different between 0.3% bile salt and 0.5% bile salt. This finding indicates that the isolates possess good resistance to 0.3% and 0.5% bile salt, as they retain their cell viability (\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe ability of these lactobacilli isolates to resist the effects of bile salt toxicity may be due to bile salt hydrolases (BSHs), bile salt efflux, and modifications in the bacterial membrane composition (\u003cspan citationid=\"CR88\" class=\"CitationRef\"\u003e88\u003c/span\u003e). BSH sticks to bile salts and decreases their toxicity (\u003cspan citationid=\"CR122\" class=\"CitationRef\"\u003e122\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eWith regards to \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e, bile salt tolerance can be regulated by inducing BSH, modification of cell membrane composition and fluidity, preventing oxidative damage, and sustaining the proton motive force (\u003cspan citationid=\"CR123\" class=\"CitationRef\"\u003e123\u003c/span\u003e). Oh \u0026amp; Jung(\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e) mentioned that several bile tolerance levels of three \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e strains were linked to six proteins (GshR1, GshR4, Cfa2, Bsh1, OpuA, and AtpH) that may be crucial to the bile salt response and adaptation in this \u003cem\u003eLactobacillus\u003c/em\u003e species. Additionally, the bsh gene expression is upregulated in \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e following bile exposure (\u003cspan citationid=\"CR124\" class=\"CitationRef\"\u003e124\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe presence of polysaccharides on their outer cell membrane (\u003cspan citationid=\"CR87\" class=\"CitationRef\"\u003e87\u003c/span\u003e), S-layer protein protection and extracellular peptidoglycan, lipoteichoic acids, teichoic acid and proteins in \u003cem\u003eLactobacillus\u003c/em\u003e support the maintenance of the organism's cellular integrity (\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eIn accordance with our study, no significant differences in the tolerance rates of \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e CCFM737 and RS4 were recorded when the bile salt concentration increased from 0% to 10% within a 12 h incubation time (\u003cspan citationid=\"CR88\" class=\"CitationRef\"\u003e88\u003c/span\u003e). Similarly, there were no significant differences observed in the growth rates of \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e strains SWO3 and SW06, after 24 h of incubation with 0.3% and 1% bile salt (\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e). Zhang et al. (\u003cspan citationid=\"CR103\" class=\"CitationRef\"\u003e103\u003c/span\u003e) reported that no significant change in the bacterial viability rate was noted for \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e incubated in 3% bile salts. \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e strains Lp 21,30,31,32,33,35,41,52, and 55 displayed no significant changes when exposed to 0.3% and 0.5% bile salt after 24 h (\u003cspan citationid=\"CR94\" class=\"CitationRef\"\u003e94\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eHowever, none of the \u003cem\u003eLactobacillus plantarum\u003c/em\u003e strains sourced from tropical-grown fruits and vegetables were resistant to 0.2% and 0.3% bile salt after 24 h of incubation (\u003cspan citationid=\"CR77\" class=\"CitationRef\"\u003e77\u003c/span\u003e). In addition, \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e and \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e strains sourced from the native chicken gut could not survive after 2 h in the presence of 0.3% (w/v) bile salt (\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe resistance of our study isolates to bile acid shows that they have good potential to survive bile salt stress conditions in the small intestine and remain viable to exert their probiotic functions (\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e).\u003c/p\u003e\u003cdiv id=\"Sec31\" class=\"Section2\"\u003e\u003ch2\u003eBSH activity\u003c/h2\u003e\u003cp\u003eBSH is an intracellular enzyme that catalyzes the hydrolysis of amide bonds between glycine or taurine and the steroid nucleus of bile salts (\u003cspan citationid=\"CR125\" class=\"CitationRef\"\u003e125\u003c/span\u003e). The ability of LAB to deconjugate bile salt is considered an indication of host-microbe interactions in the gut, which enable functional regulation of cholesterol metabolism (\u003cspan citationid=\"CR121\" class=\"CitationRef\"\u003e121\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThree isolates, \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e (To3a, tomato), \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e (Chcx3a, chicken gut), and \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e (Cu3e, cucumber), exhibited the highest BSH activity with precipitation zones of 10\u0026ndash;15 mm (++), and higher than the 1\u0026ndash;10mm (+) of \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG and other isolates.\u003c/p\u003e\u003cp\u003eBSH is commonly distributed within the human and animal GIT and mainly originates from gram-positive intestinal bacteria such as \u003cem\u003eLactobacillus\u003c/em\u003e species (\u003cspan citationid=\"CR126\" class=\"CitationRef\"\u003e126\u003c/span\u003e). Lactobacilli are capable of performing various BSH enzymatic functions in the presence of bile salts, in particular, two BSH genes (\u003cem\u003ebsh\u003c/em\u003e1 and \u003cem\u003ebsh\u003c/em\u003e3) out of 4 (\u003cem\u003ebsh\u003c/em\u003e2 and \u003cem\u003ebsh\u003c/em\u003e4) in \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e are linked with BSH activity (\u003cspan citationid=\"CR127\" class=\"CitationRef\"\u003e127\u003c/span\u003e). BSH enzymes play a vital role in reducing these toxic effects of glycoconjugated bile salts by allowing bile to enter the intestinal duodenum or microenvironments, when the pH decreases as a result of LAB (\u003cspan citationid=\"CR128\" class=\"CitationRef\"\u003e128\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eFurthermore, lactobacilli strains derived from the GIT, where bile is richly present, are more likely to be BSH positive (\u003cspan citationid=\"CR129\" class=\"CitationRef\"\u003e129\u003c/span\u003e). The overexpression and deletion of one or more \u003cem\u003ebsh\u003c/em\u003e genes may be responsible for BSH activity in lactobacilli (\u003cspan citationid=\"CR130\" class=\"CitationRef\"\u003e130\u003c/span\u003e). The higher expression of the \u003cem\u003ebsh\u003c/em\u003e3 gene in \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e is strongly associated with bile salt resistance (\u003cspan citationid=\"CR127\" class=\"CitationRef\"\u003e127\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eOur study corroborates a study that reported a large precipitation of up to 20 mm for \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e F2 and 2F8 sourced from fresh figs and approximately 15 mm for \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e FB13 derived from pricky pears against MRS agar supplemented with porcine bile (\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e). Another study observed that \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e RC (raw cheese) and \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e GV (guava) both presented precipitation zones\u0026thinsp;\u0026gt;\u0026thinsp;1.5 cm greater than those of \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG (1.0\u0026ndash;1.5 cm) (\u003cspan citationid=\"CR131\" class=\"CitationRef\"\u003e131\u003c/span\u003e). Additionally, \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e TF-7, derived from pickled olives, exhibited a strong precipitation zone\u0026thinsp;\u0026ge;\u0026thinsp;13.1 mm against the sodium salt of taurodeoxycholic (\u003cspan citationid=\"CR99\" class=\"CitationRef\"\u003e99\u003c/span\u003e). Meanwhile, intestinal \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e LR20 strain displayed an intense level of BSH activity, as shown by dense precipitation of sodium taurocholate and sodium tauroglycocholate (\u003cspan citationid=\"CR96\" class=\"CitationRef\"\u003e96\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eUnlike our findings, \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e V3F and \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG both showed precipitation zones of up to 1.0\u0026ndash;1.5 cm in a study by Boricha et al. (2019). \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e 299v and DGIA1 showed low precipitation zones of 0 and 2 mm, respectively (\u003cspan citationid=\"CR132\" class=\"CitationRef\"\u003e132\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eIn a different study, none of the \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e strains isolated from fermented cabbage or cucumber (\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e), or rotten fruits or vegetables (\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e) showed any BSH activity.\u003c/p\u003e\u003cp\u003eSome lactobacilli strains have different BSH-encoding genes; for example, \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e possesses four genes (\u003cem\u003ebsh\u003c/em\u003e 1, \u003cem\u003ebsh\u003c/em\u003e 2, \u003cem\u003ebsh\u003c/em\u003e 3, and \u003cem\u003ebsh\u003c/em\u003e 4) compared with others (\u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e, \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e and \u003cem\u003eLimosilactobacillus fermentum\u003c/em\u003e), which contain only one or two \u003cem\u003ebsh\u003c/em\u003e genes (\u003cspan citationid=\"CR125\" class=\"CitationRef\"\u003e125\u003c/span\u003e, \u003cspan citationid=\"CR130\" class=\"CitationRef\"\u003e130\u003c/span\u003e). The origin of the \u003cem\u003eLactobacillus\u003c/em\u003e strains plays an important role in BSH activity, as GIT or faecal-sourced microbes may have high BSH activity compared with bacteria sourced from plants or fermented foods (\u003cspan citationid=\"CR129\" class=\"CitationRef\"\u003e129\u003c/span\u003e, \u003cspan citationid=\"CR133\" class=\"CitationRef\"\u003e133\u003c/span\u003e). According to Kumar et al. (\u003cspan citationid=\"CR133\" class=\"CitationRef\"\u003e133\u003c/span\u003e), BSH enzymes in \u003cem\u003eLactobacillus\u003c/em\u003e isolates have different substrate specificities, considering that the majority of \u003cem\u003eBsh\u003c/em\u003e-active strains exhibit a preference for hydrolyzing glycocholate bile salt compared with taurocholate and taurodeoxycholate bile salts.\u003c/p\u003e\u003cp\u003eIsolates with positive BSH activity have good potential for use as a dietary supplement (\u003cspan citationid=\"CR134\" class=\"CitationRef\"\u003e134\u003c/span\u003e). They also efficiently colonize the GIT and could be useful in reducing cholesterol levels in the body when administered as probiotics (\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eHowever, the \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e isolate (Mcxc, chicken gut) showed no precipitation zone in this study (Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e\u003cb\u003e)\u003c/b\u003e. The reason may be due to the absence of the \u003cem\u003ebsh\u003c/em\u003e gene or its poor expression (\u003cspan citationid=\"CR130\" class=\"CitationRef\"\u003e130\u003c/span\u003e). Moreover, not all lactobacilli strains sourced from the GIT have BSH activity (\u003cspan citationid=\"CR129\" class=\"CitationRef\"\u003e129\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eConversely, BSH activity was observed for \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e 334 (\u003cspan citationid=\"CR135\" class=\"CitationRef\"\u003e135\u003c/span\u003e),\u003c/p\u003e\u003cp\u003e\u003cem\u003eLactobacillus johnsonii\u003c/em\u003e NCD01693 (\u003cspan citationid=\"CR133\" class=\"CitationRef\"\u003e133\u003c/span\u003e), \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e BFE 6128 and BFE 6154 (\u003cspan citationid=\"CR136\" class=\"CitationRef\"\u003e136\u003c/span\u003e) in a previous study.\u003c/p\u003e\u003cp\u003e\u003cem\u003eLactobacillus\u003c/em\u003e isolates with no BSH activity are unable to survive and colonize the mucosal surfaces of the small intestines and hence cannot carry out cholesterol-lowering functions (\u003cspan citationid=\"CR137\" class=\"CitationRef\"\u003e137\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cb\u003eCholesterol-lowering ability of\u003c/b\u003e \u003cb\u003eLactobacillus\u003c/b\u003e \u003cb\u003eisolates\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe cholesterol-lowering ability of \u003cem\u003eLactobacillus\u003c/em\u003e isolates is essential because it can help reduce harmful LDL cholesterol levels in the serum, thereby reducing the risk of cardiovascular disease (\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e, \u003cspan citationid=\"CR138\" class=\"CitationRef\"\u003e138\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe isolates \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e (To3a and To3d, tomato), \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e (Chx3a, Chicken gut), and \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e (Cu3e, cucumber) exhibited significantly higher cholesterol-lowering abilities (p\u0026thinsp;\u0026le;\u0026thinsp;0.05) compared with the \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG, which showed a 41.81% reduction. Among these, \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e (To3d, tomato) and \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e (Chx3a, chicken gut) demonstrated cholesterol-lowering capacities exceeding 50%, highlighting their strong potential as probiotic candidates for managing hypercholesterolemia (\u003cspan citationid=\"CR128\" class=\"CitationRef\"\u003e128\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eAccording to our findings, there was no relationship between cholesterol-lowering ability and the source of the strains investigated.\u003c/p\u003e\u003cp\u003eProbiotic \u003cem\u003eLactobacillus\u003c/em\u003e strains can generate BSH enzymes that deconjugate bile salts, resulting in reduced cholesterol reabsorption and lower solubility (\u003cspan citationid=\"CR128\" class=\"CitationRef\"\u003e128\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eErt\u0026uuml;rkmen et al. (2023) reported that these strains directly assimilate cholesterol into their cells and incorporate cholesterol into their cell membrane. This is later excreted from the body through faeces, hence lowering total cholesterol levels (\u003cspan citationid=\"CR139\" class=\"CitationRef\"\u003e139\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe assimilation of cholesterol by lactobacilli isolates lowers the quantity of intestinal cholesterol that may be absorbed by enterocytes, hence reducing cholesterol absorption as well as its overall level (\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eCholesterol binds to the lactobacilli cell surface via adsorption (\u003cspan citationid=\"CR140\" class=\"CitationRef\"\u003e140\u003c/span\u003e), leading to a smaller amount of readily absorbed cholesterol in the intestine (\u003cspan citationid=\"CR141\" class=\"CitationRef\"\u003e141\u003c/span\u003e). Choi \u0026amp; Chang (\u003cspan citationid=\"CR142\" class=\"CitationRef\"\u003e142\u003c/span\u003e) attributed the unique chemical and structural properties of \u003cem\u003eLactobacillus plantarum\u003c/em\u003e cell wall compared with those of other LAB cell types as a possible reason for its strong cholesterol attachment.\u003c/p\u003e\u003cp\u003eShort-chain fatty acids, which are mainly generated by gut microbes as a result of the fermentation of dietary fibre and resistant starch, can promote the conversion of cholesterol to bile acid, which, in turn, leads to a delayed atherosclerosis process (\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e). Moreover, EPS produced by certain LAB may bind bile acids and facilitate their excretion, which promotes the synthesis of bile acids from cholesterol, hence reducing the total circulating cholesterol (\u003cspan citationid=\"CR139\" class=\"CitationRef\"\u003e139\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eIn line with our findings, \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e strain CGMCC1.557 derived from vegetables exhibited a higher cholesterol assimilation ability (58%) than the 39% expressed by \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG (\u003cspan citationid=\"CR143\" class=\"CitationRef\"\u003e143\u003c/span\u003e). Furthermore, \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e strain NCIMB 701089 assimilated more than 67% of cholesterol in their investigation, compared with the control (no probiotic), which accounted for 0%, and \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG, which accounted for less than 50% (\u003cspan citationid=\"CR144\" class=\"CitationRef\"\u003e144\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eA higher cholesterol removal ability was observed in a previous study, where \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e NCA4 (prickly pear) and 2F8 (fresh figs) exhibited cholesterol removal abilities of 66.04% and 81.24%, respectively, compared with the control after 24 h of incubation (\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e). Compared with those of the control, cholesterol assimilation by \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e strains MYSDV5 and MYSDV2 sourced from traditional fermented food was reported to be 75.7% and 65.2% respectively (\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e isolate DLBSK207 (goat colostrum) displayed a cholesterol removal ability of 84.67%, followed by \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e isolate DLBSH122 (mango fruit), with 65.34% cholesterol removal ability (\u003cspan citationid=\"CR141\" class=\"CitationRef\"\u003e141\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eIn contrast, \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e strains DUR 2, DUR5, and DUR8 sourced from Tempoyak presented a lower cholesterol removal ability (25.99%, 60.89% and 30.37%, respectively) than did \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG (61.41%) (\u003cspan citationid=\"CR107\" class=\"CitationRef\"\u003e107\u003c/span\u003e). Moreover, \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e NR74, which was isolated from kimchi, and \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG showed similar cholesterol reduction rates of 47.8% and 48%, respectively, in a previous study (\u003cspan citationid=\"CR145\" class=\"CitationRef\"\u003e145\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe importance of \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e To3d and \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e Chx3a isolates, which exhibited a cholesterol-lowering ability of more than 50% suggests that they could be promising probiotic candidates for managing hypercholesterolemia because of their potent cholesterol-lowering effects (\u003cspan citationid=\"CR146\" class=\"CitationRef\"\u003e146\u003c/span\u003e), and their ability to minimize the risk of cardiovascular diseases (\u003cspan citationid=\"CR138\" class=\"CitationRef\"\u003e138\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec32\" class=\"Section2\"\u003e\u003ch2\u003eThe EPS production ability\u003c/h2\u003e\u003cp\u003eThe EPS production capability of some probiotic microorganisms is an attractive functional property, as it helps bacterial cells resist unfavourable environmental stress and allows effective gut colonization of mucosal surfaces (\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e), by increasing their auto-aggregation capacity (\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eMost isolates, \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e (To3a, To3b, and To3d, tomato), \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e (Chx3a, chicken gut), \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e (Cu2f, cucumber), \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e (Cu3e, cucumber) and \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e (Mcxb, chicken gut), exhibited the ability to produce EPS.\u003c/p\u003e\u003cp\u003eEPS are polymers made up of long chains of sugar molecules that have high molecular weights and differ in their properties and structures (147). They are secondary metabolites secreted into the extracellular environment during the growth and metabolism of microbes (\u003cspan citationid=\"CR148\" class=\"CitationRef\"\u003e148\u003c/span\u003e, \u003cspan citationid=\"CR149\" class=\"CitationRef\"\u003e149\u003c/span\u003e), and EPS molecules attach to the surface of the bacterial cell wall in the form of slime or a capsule (\u003cspan citationid=\"CR150\" class=\"CitationRef\"\u003e150\u003c/span\u003e). \u003cem\u003eLactobacillus\u003c/em\u003e species, including \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e, \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e, \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e and \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e, are among the best EPS producers (\u003cspan citationid=\"CR151\" class=\"CitationRef\"\u003e151\u003c/span\u003e, \u003cspan citationid=\"CR152\" class=\"CitationRef\"\u003e152\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eIn particular, species including \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e and \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e produce heteropolysaccharides (Heps) (\u003cspan citationid=\"CR153\" class=\"CitationRef\"\u003e153\u003c/span\u003e), which are synthesized within the cell and then carried out of the cell, whereas species such as \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e, \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e and \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e produce homopolysaccharides that are synthesized outside the cell, where a particular enzyme collects and assembles the sugar residues (\u003cspan citationid=\"CR154\" class=\"CitationRef\"\u003e154\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eSome proteins and enzymes participate in EPS production, which can be controlled by the regulation of gene expression of these proteins and enzymes (\u003cspan citationid=\"CR155\" class=\"CitationRef\"\u003e155\u003c/span\u003e). Pourjafar et al. (\u003cspan citationid=\"CR68\" class=\"CitationRef\"\u003e68\u003c/span\u003e) highlighted that the presence of the glycosyltransferase enzymes plays a significant role in the biosynthesis of EPS via available sugar units. Furthermore, certain gene cluster, such as \u003cem\u003eeps\u003c/em\u003e, in \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e, play key roles in EPS biosynthesis (\u003cspan citationid=\"CR148\" class=\"CitationRef\"\u003e148\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eOur results agree with the EPS production results observed for \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e isolates (SHA101, SHA103, SHA104, SHA107, SHA111 and SHA113) and \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e (SHA105) (\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e), \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e SJTUF 62116 (\u003cspan citationid=\"CR150\" class=\"CitationRef\"\u003e150\u003c/span\u003e), \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e SHA2-3B (\u003cspan citationid=\"CR156\" class=\"CitationRef\"\u003e156\u003c/span\u003e), and \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e PUMSKGRI (\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eSimilarly, \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e 53 (\u003cspan citationid=\"CR104\" class=\"CitationRef\"\u003e104\u003c/span\u003e), \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e KMUDR7 (\u003cspan citationid=\"CR121\" class=\"CitationRef\"\u003e121\u003c/span\u003e) and \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e 47FE (\u003cspan citationid=\"CR157\" class=\"CitationRef\"\u003e157\u003c/span\u003e) have been reported to show EPS production capacity.\u003c/p\u003e\u003cp\u003eThe wide range of sources of EPSs results in variation in their structural composition and physicochemical properties, and these dissimilarities confer different biological functions(\u003cspan citationid=\"CR149\" class=\"CitationRef\"\u003e149\u003c/span\u003e) Moreover, the differences in EPS may vary owing to monosaccharide conformation, the connections between builder units, charge, the existence of frequent side-chains, and replacement (\u003cspan citationid=\"CR68\" class=\"CitationRef\"\u003e68\u003c/span\u003e). In addition to the factors mentioned above, the conditions of the environment, such as pH, temperature, nutrient composition and the carbon source utilized by the bacterium (\u003cspan citationid=\"CR92\" class=\"CitationRef\"\u003e92\u003c/span\u003e, \u003cspan citationid=\"CR158\" class=\"CitationRef\"\u003e158\u003c/span\u003e), the transcriptional gene levels necessary for the production of EPS(\u003cspan citationid=\"CR159\" class=\"CitationRef\"\u003e159\u003c/span\u003e) and the over-expression of the \u003cem\u003eesp\u003c/em\u003e gene clusters in the bacterium may significantly influence EPS production capacity.\u003c/p\u003e\u003cp\u003eEPS produced by these strains may be leveraged by the fermented food industry as bio-thickeners, due to their water-binding ability, thereby improving the rheological features, and further playing an important role in food processes such as viscosity, emulsification, and flocculation (\u003cspan citationid=\"CR160\" class=\"CitationRef\"\u003e160\u003c/span\u003e). In the dairy industry, \u003cem\u003eLactobacillus\u003c/em\u003e EPS may play a primary role when used as thickeners and texturizers to enhance food product viscosity, as they may also act as stabilizers, reduce syneresis, and interact with different milk constituents (\u003cspan citationid=\"CR155\" class=\"CitationRef\"\u003e155\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eAdditionally, EPS producing \u003cem\u003eLactobacillus\u003c/em\u003e presents potential as a replacement for chemical additives (\u003cspan citationid=\"CR161\" class=\"CitationRef\"\u003e161\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eConversely, the \u003cem\u003eLactobacillus\u003c/em\u003e isolates \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e (Chx3b, chicken gut), \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e (Mcyc, chicken gut), \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e (Pb2, banana), and \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e (Mcxc, chicken gut) were unable to produce EPS, suggesting potential variability in EPS production among the strains studied.\u003c/p\u003e\u003cp\u003eThe absence or deletion of the crucial cluster gene (\u003cem\u003eesp\u003c/em\u003e), essential for EPS biosynthesis, may result in low EPS yields (\u003cspan citationid=\"CR162\" class=\"CitationRef\"\u003e162\u003c/span\u003e, \u003cspan citationid=\"CR163\" class=\"CitationRef\"\u003e163\u003c/span\u003e). A mutant strain or alterations in the \u003cem\u003eeps\u003c/em\u003e cluster genes could significantly impair the production level of EPS in the \u003cem\u003eLactobacillus\u003c/em\u003e strain (\u003cspan citationid=\"CR162\" class=\"CitationRef\"\u003e162\u003c/span\u003e, \u003cspan citationid=\"CR164\" class=\"CitationRef\"\u003e164\u003c/span\u003e). Moreover, the absence of the glycosyltransferase enzyme necessary for EPS biosynthesis may result in a lack of EPS production (\u003cspan citationid=\"CR68\" class=\"CitationRef\"\u003e68\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eOther factors, such as nutrient composition, especially starvation or an oversupply of essential nutrients such as nitrogen, sugars, carbon dioxide, etc., could alter EPS biosynthesis (\u003cspan citationid=\"CR92\" class=\"CitationRef\"\u003e92\u003c/span\u003e). For example, Fuso et al. (\u003cspan citationid=\"CR154\" class=\"CitationRef\"\u003e154\u003c/span\u003e) mentioned that a low EPS production yield in \u003cem\u003eLactobacillus\u003c/em\u003e depends on the kind of sugar (glucose, maltose, lactose, galactose, mannose, ribose, glucosamine and fructose) used as a carbon source. According to Mıdık et al. (\u003cspan citationid=\"CR165\" class=\"CitationRef\"\u003e165\u003c/span\u003e), LAB EPS production is negatively affected by an increase in temperature from 30\u0026ndash;37\u0026deg;C, extended incubation time above 48 h, pH 5.0 and 7.0 and increased NaCl concentration of 3% and 6%.\u003c/p\u003e\u003cp\u003eIn contrast to our findings, \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e PUMSKGRI (\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e) and \u003cem\u003eLactobacillus reuteri\u003c/em\u003e DUR12 (\u003cspan citationid=\"CR107\" class=\"CitationRef\"\u003e107\u003c/span\u003e) demonstrated EPS production ability.\u003c/p\u003e\u003cp\u003e\u003cem\u003eLactobacillus\u003c/em\u003e isolates with a negative EPS production capacity may play an insignificant role when used in the food industry as viscous, stabilizing and emulsifying agents (\u003cspan citationid=\"CR92\" class=\"CitationRef\"\u003e92\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cb\u003eDPPH scavenging activity of\u003c/b\u003e \u003cb\u003eLactobacillus\u003c/b\u003e \u003cb\u003estrains\u003c/b\u003e\u003c/p\u003e\u003cp\u003eAn imbalance between high levels of reactive oxygen species (ROS) and low antioxidant capacity is known as oxidative stress (\u003cspan citationid=\"CR166\" class=\"CitationRef\"\u003e166\u003c/span\u003e, \u003cspan citationid=\"CR167\" class=\"CitationRef\"\u003e167\u003c/span\u003e). ROS are generated during normal metabolic processes and play essential roles in various biological processes (\u003cspan citationid=\"CR168\" class=\"CitationRef\"\u003e168\u003c/span\u003e). However, excessive oxidative stress has been linked to several chronic conditions such as cancer, diabetes, hypertension, ulcers, aging, and atherosclerosis (\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e). Interestingly, some probiotic microbes produce bioactive compounds that can counteract oxidative stress via the prevention of ROS formation through specific molecular mechanisms (\u003cspan citationid=\"CR169\" class=\"CitationRef\"\u003e169\u003c/span\u003e). Therefore, identifying these organisms capable of exerting antioxidant activity in the human body is desirable.\u003c/p\u003e\u003cp\u003eThe DPPH scavenging activity of \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e (To3d, tomato) (24.45%) was significantly (p\u0026thinsp;\u0026le;\u0026thinsp;0.05) higher than that of \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG (23.20%), whereas the activity of the isolate \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e (Mcyc, chicken gut) was not significantly different (P\u0026thinsp;\u0026gt;\u0026thinsp;0.05).\u003c/p\u003e\u003cp\u003eThe \u003cem\u003eLactobacillus\u003c/em\u003e genus can generate antioxidant enzymes (catalase, superoxide dismutase (SOD), and nicotinamide adenine dinucleotide phosphate (NADH) oxidase and NADH peroxidase), or non-enzymatic antioxidants (Mn\u003csup\u003e2+\u003c/sup\u003e, vitamins C and E, tocoferols, glutathione), which depend on small molecule activity together with enzymes to neutralize excess ROS (\u003cspan citationid=\"CR101\" class=\"CitationRef\"\u003e101\u003c/span\u003e, \u003cspan citationid=\"CR170\" class=\"CitationRef\"\u003e170\u003c/span\u003e). For example, catalase plays a valuable role in alleviating oxidative stress by degrading hydrogen peroxide (H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e) (\u003cspan citationid=\"CR171\" class=\"CitationRef\"\u003e171\u003c/span\u003e). Lactobacilli can produce EPS and organic acids (lactic acid, acetic acid, etc.), which contribute to alleviating the effects of ROS and enhancing microbial antioxidant activity (\u003cspan citationid=\"CR172\" class=\"CitationRef\"\u003e172\u003c/span\u003e). This is due to its ability to donate electrons or hydrogen atoms, eliminate free radicals, and chelate metal ions (\u003cspan citationid=\"CR161\" class=\"CitationRef\"\u003e161\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eIn addition, the antioxidant molecules (C30 carotenoid exopolysaccharides, ferulic acid and lactate) produced by these \u003cem\u003eLactobacillus\u003c/em\u003e may also mitigate free radical formation through a neutralization reaction (\u003cspan citationid=\"CR173\" class=\"CitationRef\"\u003e173\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eCertain \u003cem\u003eLactobacillus\u003c/em\u003e species possess oxidative stress resistance genes and proteins essential for redox mechanisms, such as thioredoxin antioxidant proteins expressed by certain \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e and \u003cem\u003eLacticaseibacillus casei\u003c/em\u003e strains or the catalase gene (\u003cem\u003ekat\u003c/em\u003eA) expressed by \u003cem\u003eLatilactobacillus sakei\u003c/em\u003e (\u003cspan citationid=\"CR170\" class=\"CitationRef\"\u003e170\u003c/span\u003e). Additionally, they can efficiently prevent high levels of ROS formation and reduce ROS stress by chelating with metal ions (Fe2+) (\u003cspan citationid=\"CR172\" class=\"CitationRef\"\u003e172\u003c/span\u003e, \u003cspan citationid=\"CR174\" class=\"CitationRef\"\u003e174\u003c/span\u003e). Feng \u0026amp; Wang (\u003cspan citationid=\"CR174\" class=\"CitationRef\"\u003e174\u003c/span\u003e) highlighted that these microbes possess a ROS-binding (glutathione and thioredoxin) system that maintains intracellular dithiol/disulfide homeostasis in bacterial cells, playing a vital role in protection against oxidative stress and thereby protecting bacteria from oxidative damage.\u003c/p\u003e\u003cp\u003eThe antioxidant capacity of \u003cem\u003eLactobacillus\u003c/em\u003e isolates corresponds with findings from earlier research, where \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e strain DUR8 exhibited DPPH scavenging activity (73.36%), which was higher than that of the reference strain \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG (59.15%) (\u003cspan citationid=\"CR107\" class=\"CitationRef\"\u003e107\u003c/span\u003e). Another study revealed that \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e 27156, 27195, 27197 and 27319 displayed a maximum DPPH scavenging rates of 34.85%, 34.85%, 30.3% and 39.3%, respectively, compared with the 28.24% reported for \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG (\u003cspan citationid=\"CR175\" class=\"CitationRef\"\u003e175\u003c/span\u003e). DPPH scavenging activity was 30.51% for \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e 200655, whereas it was 27.93 % for \u003cem\u003eLacicaseibacillus rhamnosus\u003c/em\u003e GG (\u003cspan citationid=\"CR166\" class=\"CitationRef\"\u003e166\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eConversely, Kim et al. (2020) reported that the DPPH scavenging rate of \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e MG505 was significantly higher (33.5%) than that of \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e MG316 (22.2%).\u003c/p\u003e\u003cp\u003eProbiotic \u003cem\u003eLactobacillus\u003c/em\u003e organisms that have high antioxidant activity may serve as antioxidant supplements that can reduce ROS levels in the host\u0026rsquo;s body tissues and considerably reduce the risk of oxidative damage in humans and animals when ingested (\u003cspan citationid=\"CR101\" class=\"CitationRef\"\u003e101\u003c/span\u003e). Additionally, treatment with probiotic \u003cem\u003eLactobacillus\u003c/em\u003e antioxidant supplements may prevent or manage ailments, such as inflammatory bowel disease, by downregulating the expression of inflammatory factors while promoting an increase in related antioxidant enzymes, possibly through the activation of various host signalling pathways (\u003cspan citationid=\"CR170\" class=\"CitationRef\"\u003e170\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe DPPH scavenging activities of the \u003cem\u003eLactobacillus\u003c/em\u003e isolates, \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e (To3b and To3b, tomato), \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e (Chx3a, chicken gut), \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e (Chx3b, chicken gut), \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e (Cu2f, cucumber), \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e (Cu3e, cucumber), and \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e (Pb2, banana), were significantly (p\u0026thinsp;\u0026le;\u0026thinsp;0.05) lower than those of the reference strain \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG (23.20%).\u003c/p\u003e\u003cp\u003eThe antioxidant activity of \u003cem\u003eLactobacillus\u003c/em\u003e species differs as a result of dissimilarities in metabolic pathways, such as the recombination DNA repair pathway, microbial metabolism, pyruvate metabolism, etc. (\u003cspan citationid=\"CR171\" class=\"CitationRef\"\u003e171\u003c/span\u003e). According to Zhao et al. (\u003cspan citationid=\"CR170\" class=\"CitationRef\"\u003e170\u003c/span\u003e), only a few species possess oxidative stress resistance genes and proteins important for redox mechanisms. In some cases, some \u003cem\u003eLactobacillus\u003c/em\u003e strains may not possess SOD genes, suggesting that different species encode different redox\u0026ndash;related genes and have different redox systems (\u003cspan citationid=\"CR174\" class=\"CitationRef\"\u003e174\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eFurthermore, certain lactobacilli demonstrate low expression or loss of the activities of catalase and the CAT antioxidant enzyme, especially under stress conditions like pH and elevated temperatures (\u003cspan citationid=\"CR173\" class=\"CitationRef\"\u003e173\u003c/span\u003e). Prete et al. (\u003cspan citationid=\"CR176\" class=\"CitationRef\"\u003e176\u003c/span\u003e) highlighted that some microbial strains that produce low EPS may possess reduced antioxidant activity compared with high EPS producers.\u003c/p\u003e\u003cp\u003eOur results are consistent with the findings of Kim et al. (\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e), who reported that \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e MG505 had a DPPH radical scavenging rate of 33.5%, whereas \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e MG316, had a DPPH rate of 22.2%.\u003c/p\u003e\u003cp\u003eCompared with the control, ascorbic acid (37.15%), \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e MG5020 showed a lower DPPH activity of 5.32% (\u003cspan citationid=\"CR167\" class=\"CitationRef\"\u003e167\u003c/span\u003e). Vougiouklaki et al. (\u003cspan citationid=\"CR177\" class=\"CitationRef\"\u003e177\u003c/span\u003e) observed that the DPPH scavenging rate of \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e ATCC 14917 (77.53%) was lower than that of \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG (83.41%) after 210 min. Similarly, the \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e isolates LpJ2, LpJ5, LpJ6, LpJ7, LpJ8, LpJ18, and LpJ20 presented low DPPH activity that was \u0026lt;\u0026thinsp;50% after 30 min, as compared with \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG (51.52%) (\u003cspan citationid=\"CR109\" class=\"CitationRef\"\u003e109\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eDifferences in the antioxidant potential of different \u003cem\u003eLactobacillus\u003c/em\u003e strains suggest that the radical-scavenging activity of the microbes examined may be due to the strain-specific nature of \u003cem\u003eLactobacillus\u003c/em\u003e (\u003cspan citationid=\"CR178\" class=\"CitationRef\"\u003e178\u003c/span\u003e). Debnath et al.(\u003cspan citationid=\"CR109\" class=\"CitationRef\"\u003e109\u003c/span\u003e) emphasized that cell surface components, such as extracellular polysaccharides, contribute to variations in the DPPH scavenging activities of microbes. Furthermore, the difference in the scavenging rates of microbial cultures may be due to the metabolic activity of different bacterial species (\u003cspan citationid=\"CR168\" class=\"CitationRef\"\u003e168\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cem\u003eLactobacillus\u003c/em\u003e isolates with weak antioxidant activity may be less efficient at neutralizing reactive free radicals when used as probiotic antioxidant supplements (\u003cspan citationid=\"CR168\" class=\"CitationRef\"\u003e168\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cb\u003eAntimicrobial activity of\u003c/b\u003e \u003cb\u003eLactobacillus\u003c/b\u003e \u003cb\u003eisolates\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe probiotic \u003cem\u003eLactobacillus\u003c/em\u003e, which has antimicrobial activities, supports gut health by suppressing pathogens that cause diseases in the host and offers natural alternatives to antibiotics (\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe isolates, \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e (To3a, To3b, and To3d, tomato), exhibited antimicrobial activity similar to that of the reference strain \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG, with an inhibition zone of 10 to 15 mm (++) or more, against \u003cem\u003eE. coli\u003c/em\u003e, \u003cem\u003eS. aureus\u003c/em\u003e, \u003cem\u003eB. cereus\u003c/em\u003e and \u003cem\u003eL. monocytogenes\u003c/em\u003e.\u003c/p\u003e\u003cp\u003eThe high antimicrobial activity of these \u003cem\u003eLactobacillus\u003c/em\u003e strains may be attributed to the production of metabolites such as bacteriocins, H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e, and organic acids (lactic and acetic acid) that can inhibit these pathogens (\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e, \u003cspan citationid=\"CR90\" class=\"CitationRef\"\u003e90\u003c/span\u003e). For example, bacteriocins prevent pathogenic cells by disrupting their cell membrane and/or forming pores that lead to the death of the cells via a fast-acting mode of action (\u003cspan citationid=\"CR179\" class=\"CitationRef\"\u003e179\u003c/span\u003e, \u003cspan citationid=\"CR180\" class=\"CitationRef\"\u003e180\u003c/span\u003e). Notably, \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e produces organic acids that decrease the pH of the environment, consequently inhibiting the growth of pathogenic bacteria (\u003cspan citationid=\"CR116\" class=\"CitationRef\"\u003e116\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eAdditionally, they can prevent the growth of pathogenic organisms via competitive effects, enhancing the permeability of the thin outer membrane, changing the intracellular osmotic pressure, and preventing the synthesis of macromolecules (\u003cspan citationid=\"CR181\" class=\"CitationRef\"\u003e181\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eHigh EPS-producing lactobacilli strains may promote their antimicrobial activity as a result of metal chelation, inhibition of nutrient uptake via barrier formation, cell wall or cytoplasmic membrane disturbance, suppression of the synthesis of mRNA and proteins, impairing cell division or decomposing DNA (\u003cspan citationid=\"CR114\" class=\"CitationRef\"\u003e114\u003c/span\u003e). Furthermore, lactobacilli with high auto-aggregative potential can inhibit pathogenic organisms from colonizing the GIT via the formation of a barrier through auto-aggregation (\u003cspan citationid=\"CR182\" class=\"CitationRef\"\u003e182\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eBesides that, S-layer protein A (SlpA) in \u003cem\u003eLactobacillus acidophilus\u003c/em\u003e, for instance, hinders bacterial infection by blocking the activity of the cellular receptor DC-SIGN and murein hydrolase (\u003cspan citationid=\"CR183\" class=\"CitationRef\"\u003e183\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eOur findings align with those of a previous study, where the diameters of the clear zones of \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e KU200656 were reported to be 31.44 mm and 32.71 mm, against \u003cem\u003eE. coli\u003c/em\u003e and \u003cem\u003eL. monocytogenes\u003c/em\u003e, respectively, which are slightly similar to those of \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG, with inhibition zones of 29.00 mm and 23.13 mm, respectively (\u003cspan citationid=\"CR184\" class=\"CitationRef\"\u003e184\u003c/span\u003e). A previous study observed that \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e S2-5 and \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG demonstrated similar diameters of inhibition zones against \u003cem\u003eE. coli\u003c/em\u003e (2\u0026ndash;5 mm) and \u003cem\u003eS. aureus\u003c/em\u003e (\u0026lt;\u0026thinsp;2 mm) (\u003cspan citationid=\"CR185\" class=\"CitationRef\"\u003e185\u003c/span\u003e). Additionally, \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e KA18 and KLAB5 displayed the same inhibition zone as \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG at 15\u0026ndash;20 mm against \u003cem\u003eL. monocytogenes\u003c/em\u003e KACC 10764 and \u003cem\u003eS. aureus\u003c/em\u003e KCCM 40510 and 11\u0026ndash;14 mm against \u003cem\u003eE. coli\u003c/em\u003e K99 KCTC 2617 (\u003cspan citationid=\"CR186\" class=\"CitationRef\"\u003e186\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eIn another study, overnight cultures of \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e CGMCC 1.557 and \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG exhibited similar inhibition zone diameters (\u0026gt;\u0026thinsp;6 mm) against \u003cem\u003eE. coli\u003c/em\u003e, \u003cem\u003eB. cereus\u003c/em\u003e and \u003cem\u003eS. aureus\u003c/em\u003e (\u003cspan citationid=\"CR143\" class=\"CitationRef\"\u003e143\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eIn contrast, \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG exhibited robust inhibition zones of 28 mm, 27 mm, 34 mm and 29 mm against \u003cem\u003eL. monocytogenes\u003c/em\u003e, \u003cem\u003eS. aureus\u003c/em\u003e, \u003cem\u003eB. cereus\u003c/em\u003e and \u003cem\u003eE. coli\u003c/em\u003e, respectively, whereas \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e V3F presented inhibition zones of 17 mm, 25 mm, 17 mm and 40 mm against \u003cem\u003eL. monocytogenes\u003c/em\u003e, \u003cem\u003eS. aureus\u003c/em\u003e, \u003cem\u003eB. cereus\u003c/em\u003e, and \u003cem\u003eE.\u003c/em\u003e coli, respectively (\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe differences in the antimicrobial effects observed in this study could be attributed to acidic products such as lactic and acetic acids, or bacteriocins, among others (\u003cspan citationid=\"CR187\" class=\"CitationRef\"\u003e187\u003c/span\u003e). Furthermore, the disparity in results may be due to the varying sensitivities of target strains to antimicrobial compounds produced by \u003cem\u003eLactobacillus\u003c/em\u003e species and the detection methods used (\u003cspan citationid=\"CR188\" class=\"CitationRef\"\u003e188\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eOral administration of probiotic \u003cem\u003eLactobacillus\u003c/em\u003e strains with strong antagonistic effects may be used to inhibit the growth and colonization of harmful pathogenic microbes in the host GIT and fermented products (\u003cspan citationid=\"CR90\" class=\"CitationRef\"\u003e90\u003c/span\u003e). Moreover, these lactobacilli or their antimicrobial compounds are great tools that may be applied in the control of food spoilage and/or pathogenic organisms, to improve food safety and extend bio-preservation (\u003cspan citationid=\"CR189\" class=\"CitationRef\"\u003e189\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe other \u003cem\u003eLactobacillus\u003c/em\u003e isolates- \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e (Chx3a, chicken gut),\u003c/p\u003e\u003cp\u003e\u003cem\u003eLactobacillus johnsonii\u003c/em\u003e (Chx3b, chicken gut), \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e (Cu2f, cucumber), \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e (Cu3e, cucumber), \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e (Pb2, banana), \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e (Mcyc, chicken gut), and \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e (Mcxb, and Mcxc, chicken gut) exhibited lower antimicrobial activity, with inhibition zones below the 10\u0026ndash;15 mm (++) range against one or more of the tested pathogens.\u003c/p\u003e\u003cp\u003eSome lactobacilli produce few antibacterial compounds, such as H\u003csub\u003e2\u003c/sub\u003e0\u003csub\u003e2\u003c/sub\u003e and bacteriocin, thus resulting in weak inhibitory activity against pathogens (\u003cspan citationid=\"CR190\" class=\"CitationRef\"\u003e190\u003c/span\u003e). The presence or lack of specific bacteriocin genes among \u003cem\u003eLactobacillus\u003c/em\u003e strains influences their ability to inhibit a wide range of pathogens (\u003cspan citationid=\"CR188\" class=\"CitationRef\"\u003e188\u003c/span\u003e). Moreover, the antibacterial effects of any \u003cem\u003eLactobacillus\u003c/em\u003e strains are notably reduced when the culture medium or the environmental conditions are at neutral or high pH levels (\u003cspan citationid=\"CR188\" class=\"CitationRef\"\u003e188\u003c/span\u003e, \u003cspan citationid=\"CR191\" class=\"CitationRef\"\u003e191\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eSimilarly, \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e KG 12\u0026thinsp;\u0026minus;\u0026thinsp;1 exhibited no inhibition zones (0 mm) against \u003cem\u003eE. coli\u003c/em\u003e, \u003cem\u003eB. cereus\u003c/em\u003e or \u003cem\u003eS. aureus\u003c/em\u003e in their research study (\u003cspan citationid=\"CR192\" class=\"CitationRef\"\u003e192\u003c/span\u003e). \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e and \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e expressed 9.0 mm and 9.1 mm inhibition zones, respectively, against \u003cem\u003eE. coli\u003c/em\u003e D7 (\u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e). Moreover, the diameter of the inhibition zones for \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e S2-5 and S4-1 was noted to be between 2\u0026ndash;5 mm and 6\u0026ndash;8 mm, respectively, against \u003cem\u003eE. coli\u003c/em\u003e O157, while that of \u003cem\u003eS. aureus\u003c/em\u003e AC1 was observed to be 2 mm and 0 mm (\u003cspan citationid=\"CR185\" class=\"CitationRef\"\u003e185\u003c/span\u003e). Similarly, \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e (SHA 105) exhibited a 9.3 mm inhibition zone, while \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e (SHA 101, 103, 104, 107, 111 and 113) had antimicrobial activity ranging from 11.1 mm to 14.2 mm against \u003cem\u003eE. coli\u003c/em\u003e ATCC 25922 (\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eConversely, some researchers revealed that \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e P99 exhibited higher antimicrobial activity with an inhibition zone of 23 mm against \u003cem\u003eE. coli\u003c/em\u003e ATCC 8739, 16.5 mm against \u003cem\u003eB. cereus\u003c/em\u003e ATCC 11778 and \u003cem\u003eL. monocytogenes\u003c/em\u003e ATCC 7644 and 15 mm against \u003cem\u003eS. aureus\u003c/em\u003e ATCC 25923 (\u003cspan citationid=\"CR193\" class=\"CitationRef\"\u003e193\u003c/span\u003e). \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e H11 exhibited a larger inhibition zone against \u003cem\u003eE. coli\u003c/em\u003e ATCC 25922 (21.33 mm), \u003cem\u003eS. aureus\u003c/em\u003e ATCC 6538 (19.33 mm) and \u003cem\u003eL. monocytogenes\u003c/em\u003e ATCC 10403 (\u0026gt;\u0026thinsp;15 mm) (\u003cspan citationid=\"CR106\" class=\"CitationRef\"\u003e106\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe degree of pathogenic inhibition by \u003cem\u003eLactobacillus\u003c/em\u003e isolates varies due to the strain-specific nature of \u003cem\u003eLactobacillus\u003c/em\u003e (\u003cspan citationid=\"CR194\" class=\"CitationRef\"\u003e194\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cem\u003eLactobacillus\u003c/em\u003e isolates with low antibacterial activity may possess poor bacteriostatic potential and are unattractive for probiotic application in terms of preventing or reducing the growth and colonization of opportunistic intestinal microbes (\u003cspan citationid=\"CR195\" class=\"CitationRef\"\u003e195\u003c/span\u003e).\u003c/p\u003e\u003cdiv id=\"Sec33\" class=\"Section3\"\u003e\u003ch2\u003eCell auto-aggregation\u003c/h2\u003e\u003cp\u003eThe ability of the same bacterial strains to bind to themselves has been linked to enhanced probiotic survival, colonization, and pathogen exclusion, making it a desirable trait for selecting potential probiotic strains (\u003cspan citationid=\"CR87\" class=\"CitationRef\"\u003e87\u003c/span\u003e). Moreover, auto-aggregation is a probiotic feature that involves bacterial entrapment in an aggregated form, which allows for stability in the GIT, resulting in less exposure to inhospitable intestinal conditions (\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe isolates \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e (To3a, tomato), \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e (Chx3a, chicken gut), \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e (Chx3b, chicken gut), \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e (Cu2f, cucumber), and \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e (Mcyc, chicken gut) exhibited a significant (p\u0026thinsp;\u0026le;\u0026thinsp;0.05) increase in auto-aggregation (%) after 2 and 4 h incubation compared with the \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG. Additionally, the auto-aggregation (%) of these isolates was significantly (p\u0026thinsp;\u0026le;\u0026thinsp;0.05) greater than that of the reference strain after 2 and 3 h of incubation.\u003c/p\u003e\u003cp\u003eThe presence and/or high expression of cell surface proteins in \u003cem\u003eLactobacillus\u003c/em\u003e, such as the sortase-dependent protein from \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e CMPG5300(\u003cspan citationid=\"CR196\" class=\"CitationRef\"\u003e196\u003c/span\u003e) or the serine/threonine-rich protein from \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e NCIMB 8826, are contributory factors that promote high cell auto-aggregation (\u003cspan citationid=\"CR197\" class=\"CitationRef\"\u003e197\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eAuto-aggregation is associated with cell surface hydrophobicity; hence, high surface hydrophobicity in lactobacilli may facilitate their ability to self-clump or aggregate readily (\u003cspan citationid=\"CR198\" class=\"CitationRef\"\u003e198\u003c/span\u003e). Additionally, \u003cem\u003eLactobacillus\u003c/em\u003e auto-aggregation activity is due to the specific strain and not the species; as a result, \u003cem\u003eLactobacillus\u003c/em\u003e strains exhibit a wide range of auto-aggregation percentages, as seen in \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e from 29.32% to 80% (\u003cspan citationid=\"CR199\" class=\"CitationRef\"\u003e199\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eAn earlier report mentioned that a reduction in EPS production in \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e resulted in a high increase in auto-aggregation and HT29 cell adhesion (\u003cspan citationid=\"CR200\" class=\"CitationRef\"\u003e200\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cem\u003eLactobacillus\u003c/em\u003e aggregation ability may be influenced by many factors, such as the culture conditions of the bacteria, environmental conditions (time, temperature, pH value) and bacterial pre-treatment before aggregation (\u003cspan citationid=\"CR199\" class=\"CitationRef\"\u003e199\u003c/span\u003e). Auto-aggregation is dependent on environmental conditions and incubation time, as the degree of aggregation increases with increasing incubation time (\u003cspan citationid=\"CR198\" class=\"CitationRef\"\u003e198\u003c/span\u003e). According to Zawistowska-Rojek et al. (\u003cspan citationid=\"CR201\" class=\"CitationRef\"\u003e201\u003c/span\u003e), a higher aggregation rate is observed in longer bacterial cells due to their larger surface area than those with shorter cells or spherical shapes.\u003c/p\u003e\u003cp\u003eOur observations agree with the results of a previous study in which three strains of \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e LpF, LpG and LpH showed high auto-aggregation abilities of 15.8%, 20.5% and 15.4%, respectively, compared with the 13.1% reported for \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG after a 2 h incubation period (\u003cspan citationid=\"CR201\" class=\"CitationRef\"\u003e201\u003c/span\u003e). Greater cell auto-aggregation was reported for \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e DUR12 at 47.76%, \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e DUR2, DUR5 and DUR8 at 52.24%, 48.10%, and 49.88%, respectively, than for \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG (40.55%) after 5 h (\u003cspan citationid=\"CR107\" class=\"CitationRef\"\u003e107\u003c/span\u003e). Furthermore, \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e JBCC38, JBCC93, and JBCC99 exhibited auto-aggregation abilities of 37.8%, 36%, and 31.2%, respectively, which was greater than the 20.1% reported for \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG after 2 h (\u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e64\u003c/span\u003e). In their study, \u003cem\u003eLactobacillus rhamnosus\u003c/em\u003e GG auto-aggregation values were recorded as 86%, whereas \u003cem\u003eLactobacillus plantarum\u003c/em\u003e strain OR had 92% auto-aggregation percentage after 24 h (\u003cspan citationid=\"CR131\" class=\"CitationRef\"\u003e131\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eOn the contrary, \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e PF01 presented a lower auto-aggregation ability (34%) after 24 h of incubation than did \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG (36%) (\u003cspan citationid=\"CR202\" class=\"CitationRef\"\u003e202\u003c/span\u003e). A lower degree of auto-aggregation was reported for \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e strains, ranging from 6.3% to 24.0% after 3 h and 14.4% to 29.0% after 6 h (\u003cspan citationid=\"CR203\" class=\"CitationRef\"\u003e203\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eIn a different study, a similar auto-aggregation value was observed for \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e CGMCC 1.557 (34%) and \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG (34%) after a 2 h (\u003cspan citationid=\"CR143\" class=\"CitationRef\"\u003e143\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eBased on this study result, \u003cem\u003eLactobacillus\u003c/em\u003e isolates with high auto-aggregation ability may have increased potential to adhere to and colonize the host's GIT when consumed as probiotics (\u003cspan citationid=\"CR204\" class=\"CitationRef\"\u003e204\u003c/span\u003e). Additionally, the high auto-aggregation value among \u003cem\u003eLactobacillus\u003c/em\u003e species highlights the high potential of these bacteria to inhibit pathogens from colonizing the host GIT (\u003cspan citationid=\"CR161\" class=\"CitationRef\"\u003e161\u003c/span\u003e, \u003cspan citationid=\"CR205\" class=\"CitationRef\"\u003e205\u003c/span\u003e). Considering that auto-aggregation is pivotal for biofilm development, these bacteria could be exceptional probiotic candidates when incorporated into functional foods (\u003cspan citationid=\"CR194\" class=\"CitationRef\"\u003e194\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec34\" class=\"Section3\"\u003e\u003ch2\u003eCell surface hydrophobicity\u003c/h2\u003e\u003cp\u003eCell surface hydrophobicity determines the ability of probiotic \u003cem\u003eLactobacillus\u003c/em\u003e cells to interact with and adhere to surfaces such as epithelial cells, a key feature of probiotics (\u003cspan citationid=\"CR194\" class=\"CitationRef\"\u003e194\u003c/span\u003e, \u003cspan citationid=\"CR206\" class=\"CitationRef\"\u003e206\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe isolates \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e (Cu2f, cucumber), \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e (Cu3e, cucumber), and \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e (Mcyc, chicken gut) exhibited significantly higher cell surface hydrophobicity values (p\u0026thinsp;\u0026le;\u0026thinsp;0.05) than did \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG (53.33%). In contrast, the isolate \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e (Chx3b, chicken gut) displayed cell surface hydrophobicity (%) not significantly different (p\u0026thinsp;\u0026gt;\u0026thinsp;0.05) from that of \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG.\u003c/p\u003e\u003cp\u003eThe higher degree of cell surface hydrophobicity observed in the \u003cem\u003eLactobacillus\u003c/em\u003e isolates \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e (Cu2f, cucumber), \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e (Cu3e, cucumber), and \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e (Mcyc, chicken gut) can be attributed to the presence of cell surface proteins in lactobacilli, including S-layer proteins, which facilitate hydrophobic interactions with the host cell surface (\u003cspan citationid=\"CR207\" class=\"CitationRef\"\u003e207\u003c/span\u003e). For example, the cell surface mucus-\u003c/p\u003e\u003cp\u003ebinding protein (\u003cem\u003eMapA\u003c/em\u003e) found in \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e (\u003cspan citationid=\"CR208\" class=\"CitationRef\"\u003e208\u003c/span\u003e), and the surface protein found in \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e HC-2 (\u003cspan citationid=\"CR209\" class=\"CitationRef\"\u003e209\u003c/span\u003e). In addition, an increase in the rate of cell surface hydrophobicity has been linked to certain lactobacilli producing highly hydrophobic EPS, which results in increased biofilm formation (\u003cspan citationid=\"CR210\" class=\"CitationRef\"\u003e210\u003c/span\u003e).\u003c/p\u003e\u003cp\u003ePrevious researchers have noted a similar trend of broad variations in hydrophobicity values among the \u003cem\u003eLactobacillus\u003c/em\u003e strains studied (\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e, \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e, \u003cspan citationid=\"CR95\" class=\"CitationRef\"\u003e95\u003c/span\u003e, \u003cspan citationid=\"CR104\" class=\"CitationRef\"\u003e104\u003c/span\u003e, \u003cspan citationid=\"CR211\" class=\"CitationRef\"\u003e211\u003c/span\u003e, \u003cspan citationid=\"CR212\" class=\"CitationRef\"\u003e212\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eIn agreement with our findings, 15 \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e isolates (LpJ1-7, LpJ10-13, and LpJ16-20) displayed higher xylene surface hydrophobicity (19.58% to 62.59%) than did \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG (15.96%) after 30 min of incubation (\u003cspan citationid=\"CR109\" class=\"CitationRef\"\u003e109\u003c/span\u003e). The n-hexadecane hydrophobicity values observed for 9 \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e isolates (Fb, RC, GV, OR, SG, CK, GP, HB, and OP) were reported to range from 2% to 27%, whereas \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG exhibited 13% hydrophobicity in a previous study (\u003cspan citationid=\"CR131\" class=\"CitationRef\"\u003e131\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eHydrophobicity values of 40% were recorded for \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e I2 from the poultry GIT (\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e), 61.15% for \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e L-3 sourced from the chicken gut (\u003cspan citationid=\"CR112\" class=\"CitationRef\"\u003e112\u003c/span\u003e), and 76% for \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e K18 (colostrum) (\u003cspan citationid=\"CR203\" class=\"CitationRef\"\u003e203\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe wide differences in the cell surface hydrophobicity of the \u003cem\u003eLactobacillus\u003c/em\u003e isolates in this study may be due to variations in the level of expression of cell surface proteins among lactobacilli strains and environmental conditions, which could impact the expression of surface proteins (\u003cspan citationid=\"CR106\" class=\"CitationRef\"\u003e106\u003c/span\u003e, \u003cspan citationid=\"CR109\" class=\"CitationRef\"\u003e109\u003c/span\u003e). In addition, the bacterial cell growth phase may contribute to affecting the surface hydrophobicity of these bacteria (\u003cspan citationid=\"CR90\" class=\"CitationRef\"\u003e90\u003c/span\u003e). Other compounds (core oligosaccharides, lipoteichoic acid, lipids, surface fibrils and several fimbriae) contribute to differences in the cell surface hydrophobicity (\u003cspan citationid=\"CR198\" class=\"CitationRef\"\u003e198\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe high cell surface hydrophobicity of some bacterial isolates in this study could play a pivotal role in promoting the adhesion and colonization of lactobacilli to the epithelium and mucosal surfaces of the host (\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e, \u003cspan citationid=\"CR106\" class=\"CitationRef\"\u003e106\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe low cell surface hydrophobicity (%) of the \u003cem\u003eLactobacillus\u003c/em\u003e isolates, \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e (To3a, To3b and To3d, tomato), \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e (Chx3a, chicken gut), \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e (Pb2, banana), and \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e (Mcxb and Mcxc, chicken gut) may be attributed to the absence of relevant structural and compositional features of their cell surfaces (S-proteins, adhesins, and polysaccharides), which reduce their ability to interact with hydrophobic substrates (\u003cspan citationid=\"CR213\" class=\"CitationRef\"\u003e213\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe surface proteins of several lactobacilli, including \u003cem\u003eLactobacillus crispatus\u003c/em\u003e and \u003cem\u003eLactobacillus acidophilus\u003c/em\u003e, whose ability to bind to host epithelial cells decreases after removal or disruption of the S-layer proteins (\u003cspan citationid=\"CR214\" class=\"CitationRef\"\u003e214\u003c/span\u003e). The abundance and presence of the EPS layer at the bacterial cell surface, which completely covers the cell and shields surface-bound proteins, could lower the hydrophobicity of lactobacilli (\u003cspan citationid=\"CR200\" class=\"CitationRef\"\u003e200\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe low percentage of cell surface hydrophobicity exhibited by lactobacilli strains in this study aligns with the observed xylene hydrophobicity value for 10 \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e strains (PL103, PL110, PL111, PL112, PL113, PL114, PL115, PL116, PL118, and PL120) and one \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e strain (PL105) derived from the native chicken GIT, which was recorded to be \u0026lt;\u0026thinsp;20% (\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e). An earlier study revealed that the xylene cell surface hydrophobicity exhibited by \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e BSF206 were less than 30% (\u003cspan citationid=\"CR215\" class=\"CitationRef\"\u003e215\u003c/span\u003e). A hydrophobic index of less than 40% was observed for \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e B20, whereas more than 50% hydrophobic indices were recorded for \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG (\u003cspan citationid=\"CR90\" class=\"CitationRef\"\u003e90\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cem\u003eLactobacillus\u003c/em\u003e isolate with a low percentage of cell surface hydrophobicity may not effectively colonize and persist in the host GIT when they are used as probiotics (\u003cspan citationid=\"CR216\" class=\"CitationRef\"\u003e216\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\n\u003ch3\u003eAntibiotic susceptibility\u003c/h3\u003e\n\u003cp\u003eOwing to the possibility of the transfer of antibiotic resistance genes to intestinal pathogenic bacteria, serious safety concerns regarding probiotics sheltering antibiotic resistance genes still exist (\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e, \u003cspan citationid=\"CR87\" class=\"CitationRef\"\u003e87\u003c/span\u003e), because of the potential for drug resistance in pathogenic organisms (\u003cspan citationid=\"CR181\" class=\"CitationRef\"\u003e181\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eAll the \u003cem\u003eLactobacillus\u003c/em\u003e isolates were susceptible to antibiotics: penicillin G (10 \u0026micro;g), chloramphenicol (30 \u0026micro;g), erythromycin (15 \u0026micro;g), and ampicillin (10 \u0026micro;g). The probable reason may be that most \u003cem\u003eLactobacillus\u003c/em\u003e species lack β-lactamase enzymes, which are sensitive to some of these antibiotics (\u003cspan citationid=\"CR217\" class=\"CitationRef\"\u003e217\u003c/span\u003e). The thick peptidoglycan layer in these bacterial cell walls is vulnerable to antibiotics, such as penicillin and ampicillin (\u003cspan citationid=\"CR218\" class=\"CitationRef\"\u003e218\u003c/span\u003e). Additionally, the absence of a resistance gene in some lactobacilli strains (such as the gene \u003cem\u003emec\u003c/em\u003eA, which is responsible for penicillin resistance; the chloramphenicol resistance gene \u003cem\u003ecat\u003c/em\u003e; or the erythromycin resistance gene \u003cem\u003eerm\u003c/em\u003eB) contributes to vulnerability to these antibiotics (\u003cspan citationid=\"CR217\" class=\"CitationRef\"\u003e217\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe tolerance of \u003cem\u003eLactobacillus\u003c/em\u003e species to these antibiotics is consistent with the findings of these previous studies (\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e, \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e, \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e, \u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e, \u003cspan citationid=\"CR219\" class=\"CitationRef\"\u003e219\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe high susceptibility of our \u003cem\u003eLactobacillus\u003c/em\u003e isolates to these antibiotics suggests they are safe for use as probiotics, since they may not contribute to the spread of antibiotic resistance genes in the host GIT microbiota (\u003cspan citationid=\"CR161\" class=\"CitationRef\"\u003e161\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eConversely, all the \u003cem\u003eLactobacillus\u003c/em\u003e isolates were resistant to aminoglycoside antibiotics: kanamycin (30 \u0026micro;g), gentamicin (10 \u0026micro;g), and streptomycin (10 \u0026micro;g) (Table\u0026nbsp;\u003cspan refid=\"Tab10\" class=\"InternalRef\"\u003e10\u003c/span\u003e). This may be due to the presence of aminoglycoside resistance genes such as the \u003cem\u003eaph(3\u0026prime;)\u003c/em\u003e-\u003cem\u003eIIIa\u003c/em\u003e gene, which is responsible for kanamycin resistance; the \u003cem\u003eant\u003c/em\u003e(\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e)-\u003cem\u003eIa\u003c/em\u003e (\u003cem\u003eaadE\u003c/em\u003e) gene, which is involved in streptomycin resistance; and the \u003cem\u003eant(4\u0026prime;)\u003c/em\u003e-\u003cem\u003eIa\u003c/em\u003e gene, which is responsible for neomycin resistance in lactobacilli strains (\u003cspan citationid=\"CR220\" class=\"CitationRef\"\u003e220\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eAs mentioned in an earlier study, the ability of the \u003cem\u003eLactobacillus\u003c/em\u003e genus to resist these antibiotics may also be attributed to the low impermeability of its cell surface to aminoglycoside antibiotics such as kanamycin (\u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e70\u003c/span\u003e). Moreover, the lack of cytochrome-mediated electron transport, which promotes drug absorption, is another reason why these bacteria may be intrinsically resistant to this group of antibiotics (\u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e70\u003c/span\u003e, \u003cspan citationid=\"CR87\" class=\"CitationRef\"\u003e87\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThis finding is in accordance with some studies (\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e, \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e, \u003cspan citationid=\"CR181\" class=\"CitationRef\"\u003e181\u003c/span\u003e, \u003cspan citationid=\"CR211\" class=\"CitationRef\"\u003e211\u003c/span\u003e, \u003cspan citationid=\"CR221\" class=\"CitationRef\"\u003e221\u003c/span\u003e) reporting that \u003cem\u003eLactobacillus\u003c/em\u003e strains are resistant to aminoglycoside antibiotics.\u003c/p\u003e\u003cp\u003eCell culture conditions, the position of the resistance gene, and the participation of other mechanisms might affect the degree of antibiotic resistance (\u003cspan citationid=\"CR182\" class=\"CitationRef\"\u003e182\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cem\u003eLactobacillus\u003c/em\u003e strains may survive in the gut even after treatment with aminoglycoside antibiotics and restore the intestinal microflora (\u003cspan citationid=\"CR222\" class=\"CitationRef\"\u003e222\u003c/span\u003e), and when they are co-administered with an antibiotic, they might reduce the gut side effects linked with the antibiotics (\u003cspan citationid=\"CR69\" class=\"CitationRef\"\u003e69\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eIn this present study, 6 \u003cem\u003eLactobacillu\u003c/em\u003es isolates, \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e (Chx3a, chicken gut), \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e (Chx3b, chicken gut), \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e (Pb2, banana), \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e (Mcyc, chicken gut), \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e (Mcxb and Mcxc, chicken gut), were resistant to tetracycline (Table\u0026nbsp;\u003cspan refid=\"Tab10\" class=\"InternalRef\"\u003e10\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe high resistance rate to tetracycline among chicken gut samples (Chcx3a, Chcx3b, Mcyc, Mcxb, and Mcxc) in this study may be due to the common use of tetracycline antibiotics by poultry farmers in the Johannesburg metropolitan city in South Africa. Motey et al. (\u003cspan citationid=\"CR223\" class=\"CitationRef\"\u003e223\u003c/span\u003e) highlighted that \u003cem\u003eLactobacillus\u003c/em\u003e species, which are resistant to tetracycline, may harbour transferable \u003cem\u003etet\u003c/em\u003e genes such as \u003cem\u003etet\u003c/em\u003e (\u003cem\u003eM\u003c/em\u003e) and \u003cem\u003etet\u003c/em\u003e (\u003cem\u003eW\u003c/em\u003e), which have been reported in other studies of lactobacilli.\u003c/p\u003e\u003cp\u003eTetracycline resistance among lactobacilli may occur via ribosomal protection proteins, efflux pumps and direct enzymatic inactivation (\u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e70\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eSome studies have revealed that all \u003cem\u003eLactobacillus\u003c/em\u003e isolates examined in their study were resistant to tetracycline (\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e, \u003cspan citationid=\"CR86\" class=\"CitationRef\"\u003e86\u003c/span\u003e). Furthermore, tetracycline resistance has been observed in studies of \u003cem\u003eLactobacillus\u003c/em\u003e isolates sourced from fermented cabbage and cucumber (\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e), rectal swabs of 21-day-old chickens, their faecal and feed samples (\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eOur findings imply that the tetracycline-resistant \u003cem\u003eLactobacillus\u003c/em\u003e isolates may be resistant to these antibiotics when used for probiotics, as they encourage the spread of antibiotic resistance genes in the gut microbiota and the environment (\u003cspan citationid=\"CR217\" class=\"CitationRef\"\u003e217\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eMost of the isolates (66.7%) in this study were resistant to vancomycin (Table\u0026nbsp;\u003cspan refid=\"Tab10\" class=\"InternalRef\"\u003e10\u003c/span\u003e). The resistance of \u003cem\u003eLactobacillus\u003c/em\u003e strains to vancomycin is intrinsic and non-transferable due to the absence of D-Ala-D-Lac target modification (\u003cspan citationid=\"CR161\" class=\"CitationRef\"\u003e161\u003c/span\u003e). The presence of the vancomycin-resistance gene \u003cem\u003evan\u003c/em\u003eX (\u003cspan citationid=\"CR217\" class=\"CitationRef\"\u003e217\u003c/span\u003e, \u003cspan citationid=\"CR218\" class=\"CitationRef\"\u003e218\u003c/span\u003e), which encodes D-Ala-D-Ala dipeptidase, which is crucial for cell wall synthesis, is also a contributing factor (\u003cspan citationid=\"CR224\" class=\"CitationRef\"\u003e224\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe absence of a specific drug target or cell wall impermeability also contributes to their intrinsic resistance (\u003cspan citationid=\"CR224\" class=\"CitationRef\"\u003e224\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThis finding was closely consistent with a study by Yuksekdag et al. (\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e), which reported that eleven out of eighteen \u003cem\u003eLactobacillus\u003c/em\u003e strains isolated from the chicken GIT were resistant to vancomycin. Nonetheless, some researchers have reported a relatively high incidence of vancomycin resistance among \u003cem\u003eLactobacillus\u003c/em\u003e strains (\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e, \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e, \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e, \u003cspan citationid=\"CR211\" class=\"CitationRef\"\u003e211\u003c/span\u003e, \u003cspan citationid=\"CR221\" class=\"CitationRef\"\u003e221\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eVancomycin resistance among \u003cem\u003eLactobacillus\u003c/em\u003e isolates may assist in the prevention of antibiotic-associated diarrhoea, and in some cases, it may restore gut microflora (\u003cspan citationid=\"CR224\" class=\"CitationRef\"\u003e224\u003c/span\u003e).\u003c/p\u003e\n\u003ch3\u003eBlood haemolysis\u003c/h3\u003e\n\u003cp\u003e\u003cem\u003eLactobacillus\u003c/em\u003e strains intended for probiotic use must not cause lysis of red blood cells; thus, the absence of haemolytic activity is a critical safety requirement for probiotic strains (\u003cspan citationid=\"CR225\" class=\"CitationRef\"\u003e225\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eAs observed in this study, none of the isolates showed haemolysis on blood agar. This may be attributed to the absence of haemolysin-encoding genes, which are responsible for red blood lysis in bacterial cells (\u003cspan citationid=\"CR179\" class=\"CitationRef\"\u003e179\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eSome \u003cem\u003eLactobacillus\u003c/em\u003e strains may harbour a gene encoding protein that may imitate haemolysins (\u003cspan citationid=\"CR179\" class=\"CitationRef\"\u003e179\u003c/span\u003e), and the environmental conditions (oxidative stress, low pH, and temperature), as well as the growth medium used, may result in discrepancies in some study findings (\u003cspan citationid=\"CR87\" class=\"CitationRef\"\u003e87\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eOur results corroborate with many studies that have confirmed that \u003cem\u003eLactobacillus\u003c/em\u003e is non-haemolytic (\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e, \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e, \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e, \u003cspan citationid=\"CR104\" class=\"CitationRef\"\u003e104\u003c/span\u003e, \u003cspan citationid=\"CR226\" class=\"CitationRef\"\u003e226\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eAll the isolates tested met the safety criteria for probiotic use; therefore, they are safe for human consumption as probiotics (\u003cspan citationid=\"CR161\" class=\"CitationRef\"\u003e161\u003c/span\u003e).\u003c/p\u003e\u003cdiv id=\"Sec37\" class=\"Section2\"\u003e\u003ch2\u003eDNase activity\u003c/h2\u003e\u003cp\u003eDNases are extracellular endonucleases that cleave the phosphodiester bond in the backbone of DNA, releasing free nucleotides and phosphates, thus disrupting cell functionality (\u003cspan citationid=\"CR227\" class=\"CitationRef\"\u003e227\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eNone of the examined \u003cem\u003eLactobacillus\u003c/em\u003e isolates demonstrated DNase activity, indicating a lack of potential for nucleic acid degradation and associated pathogenicity (\u003cspan citationid=\"CR228\" class=\"CitationRef\"\u003e228\u003c/span\u003e), as shown in Table\u0026nbsp;\u003cspan refid=\"Tab11\" class=\"InternalRef\"\u003e11\u003c/span\u003e.\u003c/p\u003e\u003cp\u003eThese results agree with studies that reported that \u003cem\u003eLactobacillus\u003c/em\u003e strains showed no DNase activity (\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e, \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e, \u003cspan citationid=\"CR226\" class=\"CitationRef\"\u003e226\u003c/span\u003e). Hence, this finding proves the safety of these \u003cem\u003eLactobacillus\u003c/em\u003e strains for human use as probiotics in the food industry, considering that the degradation of host DNA is often associated with virulence in pathogenic bacteria (\u003cspan citationid=\"CR228\" class=\"CitationRef\"\u003e228\u003c/span\u003e).\u003c/p\u003e\u003cdiv id=\"Sec38\" class=\"Section3\"\u003e\u003ch2\u003eCaco-2 cells adhesion capacity\u003c/h2\u003e\u003cp\u003ePotential probiotic \u003cem\u003eLactobacillus\u003c/em\u003e species must adhere to and colonize the mucosal surface and epithelial cells to ensure prolonged persistence in the GIT and exert beneficial health effects (\u003cspan citationid=\"CR117\" class=\"CitationRef\"\u003e117\u003c/span\u003e, \u003cspan citationid=\"CR204\" class=\"CitationRef\"\u003e204\u003c/span\u003e). Accordingly, Caco-2 cells are widely used in vitro to assess probiotic adhesion capacity due to their close resemblance to mature intestinal epithelial cells (\u003cspan citationid=\"CR86\" class=\"CitationRef\"\u003e86\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cem\u003eLactobacillus johnsonii\u003c/em\u003e (Chx3b, chicken gut) and \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e (To3d, tomato) exhibited higher Caco-2 cells adhesion capacities (15.29% and 12.87%, respectively) than \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG (11.40%), whereas isolate \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e (Mcyc, chicken gut) presented a slightly lower adhesion capacity of 10.53% (Table\u0026nbsp;\u003cspan refid=\"Tab12\" class=\"InternalRef\"\u003e12\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe reason for this can be attributed to the structural differences and cell wall constituents (polysaccharides, fimbriae or pili, adhesins, surface proteins, and other components) of \u003cem\u003eLactobacillus\u003c/em\u003e species, which increase their ability to attach strongly to the host epithelial cells (\u003cspan citationid=\"CR203\" class=\"CitationRef\"\u003e203\u003c/span\u003e, \u003cspan citationid=\"CR216\" class=\"CitationRef\"\u003e216\u003c/span\u003e). As mentioned previously, the surface EF-Tu moonlighting protein of \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e 423 (\u003cspan citationid=\"CR183\" class=\"CitationRef\"\u003e183\u003c/span\u003e) and the mucus-binding protein of \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG promotes high cell adhesion ability (\u003cspan citationid=\"CR225\" class=\"CitationRef\"\u003e225\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe high auto-aggregation ability of lactobacilli has also been linked to their high capacity to adhere to epithelial cells and prevent the colonization of pathogens in the GIT (\u003cspan citationid=\"CR161\" class=\"CitationRef\"\u003e161\u003c/span\u003e, \u003cspan citationid=\"CR200\" class=\"CitationRef\"\u003e200\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eMoreover, \u003cem\u003eLactobacillus\u003c/em\u003e strains with high surface hydrophobicity values have a greater capacity to adhere to epithelial cells (\u003cspan citationid=\"CR106\" class=\"CitationRef\"\u003e106\u003c/span\u003e). In addition, EPS or lipoteichoic acids produced on the cell walls of lactobacilli contain adhesive molecules, which can enhance their adhesion capacity to the intestinal epithelium (\u003cspan citationid=\"CR90\" class=\"CitationRef\"\u003e90\u003c/span\u003e). In some instances, certain \u003cem\u003eLactobacillus\u003c/em\u003e can aggregate to form a biofilm that is favourable to intestinal cell adhesion and colonization (\u003cspan citationid=\"CR199\" class=\"CitationRef\"\u003e199\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eIn line with our findings, \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e isolates KA18, KA19, KII1, KII2 and KLAB5 derived from fermented radish displayed a high adherence rate to Caco-2 cells, ranging from 25% to 49%, in contrast to those of \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG, whose adhesion capacity was reported to be 12% (\u003cspan citationid=\"CR186\" class=\"CitationRef\"\u003e186\u003c/span\u003e). The Caco-2 adhesion rate of \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e S2-5 (12 %) ad S4-1 (8.6 %) iolated from Chinese sauerkraut was higher than that of \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG (6.1 %) (\u003cspan citationid=\"CR185\" class=\"CitationRef\"\u003e185\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eIn a different study, \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e NCUA063008 and \u003cem\u003eLactobacillus plantarum\u003c/em\u003e sourced from the faeces of wild boars, native pigs and commercial pigs demonstrated a capacity of 20.16% and 17.19% respectively, to Caco-2 cells (\u003cspan citationid=\"CR86\" class=\"CitationRef\"\u003e86\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eHowever, a higher adhesion capacity to Caco-2 cells than that observed in our study was demonstrated by \u003cem\u003eLactobacillus reuteri\u003c/em\u003e SHA 113 and \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e SHA105 (90%) (\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e), and \u003cem\u003eLactobacillus reuteri\u003c/em\u003e LR21 and LR 42 (\u0026gt;\u0026thinsp;70%) isolated from the poultry gut (\u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e55\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eBacterial adhesion is a strain-specific characteristic that is strongly influenced by origin, variations in the expression levels of surface proteins, and environmental factors (\u003cspan citationid=\"CR207\" class=\"CitationRef\"\u003e207\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eHighly adherent strains may serve as good probiotic candidates for imparting beneficial health effects, including immunomodulation and the exclusion of pathogens via competition for binding sites and nutrients in the host intestinal mucosa (\u003cspan citationid=\"CR205\" class=\"CitationRef\"\u003e205\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eIn contrast, the isolates \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e (Pb2, banana) and \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e (Cu3e, cucumber) demonstrated the lowest adhesion capacities, at 2.60% and 3.58%, respectively (Table\u0026nbsp;\u003cspan refid=\"Tab12\" class=\"InternalRef\"\u003e12\u003c/span\u003e). The reason for this may be attributed to the loss or absence of the S-layer protein from the bacterial surface through chemical means or the shielding of the layer by other molecules during prolonged cultivation (\u003cspan citationid=\"CR229\" class=\"CitationRef\"\u003e229\u003c/span\u003e). Certain proteins in \u003cem\u003eLactobacillus\u003c/em\u003e species can bind weakly (through noncovalent interactions) to surface components of intestinal cells (\u003cspan citationid=\"CR90\" class=\"CitationRef\"\u003e90\u003c/span\u003e). Moreover, the EPS layers produced by lactobacilli can block the adhesion of probiotic membrane proteins to the GIT cells (\u003cspan citationid=\"CR122\" class=\"CitationRef\"\u003e122\u003c/span\u003e). As stated in a previous study, bacterial species with a poor hydrophobic index exhibit decreased cell adhesion (\u003cspan citationid=\"CR203\" class=\"CitationRef\"\u003e203\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eSimilarly, \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e JBCC38 and JBCC99 expressed a lower Caco-2 cell adhesion percentage (\u0026lt;\u0026thinsp;2%), in contrast to the 10% recorded for \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG (\u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e64\u003c/span\u003e). The adhesion capacity for \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e CS23 was less than 20%, a contrast from \u0026gt;\u0026thinsp;25% recorded for \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG (\u003cspan citationid=\"CR230\" class=\"CitationRef\"\u003e230\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eIn contrast to this study, \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e BSF206 demonstrated an adhesion percentage of 13.2%, which was higher than the percentage of \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG (6.1%) (\u003cspan citationid=\"CR215\" class=\"CitationRef\"\u003e215\u003c/span\u003e). The ability of \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e DN3 to adhere to Caco-2 cells was 16% greater than that of \u003cem\u003eLacticaseibacillus rhamnosus\u003c/em\u003e GG at 7% (\u003cspan citationid=\"CR231\" class=\"CitationRef\"\u003e231\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eDiscrepancies in the adhesion capacity of \u003cem\u003eLactobacillus\u003c/em\u003e strain to the host may be due to differences in transpeptidase-dependent proteins, and the specific binding of bacterial surface proteins, pili, peptidoglycans, and lipopolysaccharides to surface receptors of epithelial cells (\u003cspan citationid=\"CR215\" class=\"CitationRef\"\u003e215\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eProbiotic strains with poor adhesion capacity may be unable to effectively adhere to and colonize the host mucosal surfaces to confer health benefits to the host (\u003cspan citationid=\"CR183\" class=\"CitationRef\"\u003e183\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cb\u003eCPP of\u003c/b\u003e \u003cb\u003eLactobacillus\u003c/b\u003e \u003cb\u003eisolates\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe CPP is an important criterion for the validation of the characteristics of probiotic microorganisms (\u003cspan citationid=\"CR232\" class=\"CitationRef\"\u003e232\u003c/span\u003e). In this study, the CPP of three isolates, \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e (To3a and To3d, tomato) and \u003cem\u003eLatilactobacillus curvatus\u003c/em\u003e (Cu2f, cucumber), was higher (87.50%) (Table\u0026nbsp;\u003cspan refid=\"Tab13\" class=\"InternalRef\"\u003e13\u003c/span\u003e) than commercially available probiotic preparations, whose probiotic score is in a range of 75\u0026ndash;85% (\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e, \u003cspan citationid=\"CR233\" class=\"CitationRef\"\u003e233\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eConversely, a high CPP of 100% was recorded for both \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e LMEM1002 and LMEM1006 sourced from idli batter samples (\u003cspan citationid=\"CR234\" class=\"CitationRef\"\u003e234\u003c/span\u003e). \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e NKUST 817, 828, and 851 isolated from fermented food (\u003cspan citationid=\"CR233\" class=\"CitationRef\"\u003e233\u003c/span\u003e) and \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e PUMSKGRI, isolated from a homemade fermented food product, all had a CPP of 91.67% (\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eNotably, these two isolates, \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e (To3a and To3d, tomato), met the standard for a good probiotic considering their probiotic characteristics, especially acid and bile tolerance ability.\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e"},{"header":"Conclusions","content":"\u003cp\u003eThis research study presented eighteen \u003cem\u003eLactobacillus\u003c/em\u003e isolates sourced from different food sources.\u0026nbsp;\u003cem\u003eLactobacillus plantarum\u003c/em\u003e To3a and To3d isolates demonstrated promising candidates for various\u0026nbsp;probiotic\u0026nbsp;applications, based on their tolerance to acid and bile salt, cell adhesion capacity, bile salt and cholesterol-lowering ability, and strong inhibition towards pathogenic bacteria.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eIn addition, results on safety aspects revealed that they are safe for human consumption when used as probiotics. The cumulative probiotic potential of To3a and To3d isolates suggests that they may represent a good probiotic candidate essential for the development of novel local functional food products with health-promoting effects.\u003c/p\u003e\n\u003cp\u003eNevertheless, further comprehensive studies on the technological properties, as well as in vivo studies, should be considered to gain a better insight into their beneficial attributes.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eData availability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe datasets generated and/or analyzed during this study are available from the corresponding author upon reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors would like to thank the National Research Foundation (NRF) of South Africa for their financial support (Grant number: 129095). We would also like to thank the Department of Life and Consumer Sciences, University of South Africa, for their support in providing laboratory supplies.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis work is based on research supported by the National Research Foundation (NRF) of South Africa (Grant number: 129095).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors information\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAuthors and affiliations\u003c/p\u003e\n\u003cp\u003eDepartment of Life and Consumer Sciences,University of South Africa, Cnr Christiaan de Wet Road and Pioneer Avenue, Florida, Roodepoort 1710, South Africa\u003c/p\u003e\n\u003cp\u003eChioma Vivian Asiegbu\u003c/p\u003e\n\u003cp\u003eDepartment of Life and Consumer Sciences,University of South Africa, Cnr Christiaan de Wet Road and Pioneer Avenue, Florida, Roodepoort 1710, South Africa\u003c/p\u003e\n\u003cp\u003eFrederick Tawi Tabit\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eContributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eACV and FTT participated in the conceptualization and design of the study; ACV drafted the original manuscript; ACV collected the sample and data; ACV performed the experiments; ACV and FTT analyzed the data; FTT supervised; ACV and FTT wrote- reviewed \u0026amp; edited; FTT read and approved the final manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCorresponding author\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eCorrespondence to Frederick Tawi Tabit\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research protocol was approved by the College of Agriculture and Environmental Science (CAES) Health Research Ethics Committee at the University of South Africa.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no competing interests.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eClinical trial number\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDeclaration of conflicting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no potential conflicts of interest regarding the research, authorship, and/or publication of the research article.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eChoksket S, Sharma S, Harshvardhan, Pal V, Jain A, Patil PB et al. 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Futur J Pharm Sci. 2023;9(1).\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"},{"header":"Tables","content":"\u003cp\u003e\u003cstrong\u003eTable 1\u003c/strong\u003e: \u003cem\u003eLactobacillus\u003c/em\u003e species identified from chicken gut, tomatoes, cucumber and banana\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 26.9231%;\"\u003e\n \u003cp\u003eFood source\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 39.7436%;\"\u003e\n \u003cp\u003eFrequency of \u003cem\u003eLactobacillus\u003c/em\u003e species identified\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 33.3333%;\"\u003e\n \u003cp\u003eFrequency\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 26.9231%;\"\u003e\n \u003cp\u003eChicken gut\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 39.7436%;\"\u003e\n \u003cp\u003e\u003cem\u003eLactobacillus johnsonii\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003e\u003cem\u003eLimosilactobacillus\u003c/em\u003e\u003cem\u003e\u0026nbsp;reuteri\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003e\u003cem\u003eLatilactobacillus\u003c/em\u003e\u003cem\u003e\u0026nbsp;curvatus\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003e\u003cem\u003eLatilactobacillus\u003c/em\u003e\u003cem\u003e\u0026nbsp;sakei\u0026nbsp;\u003c/em\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003cem\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 33.3333%;\"\u003e\n \u003cp\u003e(n=7; 38.9%)\u003c/p\u003e\n \u003cp\u003e(n=2; 11.1%)\u003c/p\u003e\n \u003cp\u003e(n=2; 11.1%)\u003c/p\u003e\n \u003cp\u003e(n=1; 5.6%)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 26.9231%;\"\u003e\n \u003cp\u003eTomato\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 39.7436%;\"\u003e\n \u003cp\u003e\u003cem\u003eLactiplantibacillus\u003c/em\u003e\u003cem\u003e\u0026nbsp;plantarum\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 33.3333%;\"\u003e\n \u003cp\u003e(n=3; 16.6%)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 26.9231%;\"\u003e\n \u003cp\u003eCucumber\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 39.7436%;\"\u003e\n \u003cp\u003e\u003cem\u003eLactiplantibacillus\u003c/em\u003e\u003cem\u003e\u0026nbsp;plantarum\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003e\u003cem\u003eLatilactobacillus\u003c/em\u003e\u003cem\u003e\u0026nbsp;curvatus\u0026nbsp;\u003c/em\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 33.3333%;\"\u003e\n \u003cp\u003e(n=1; 5.6%)\u003c/p\u003e\n \u003cp\u003e(n=1; 5.6%)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 26.9231%;\"\u003e\n \u003cp\u003eBanana\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 39.7436%;\"\u003e\n \u003cp\u003e\u003cem\u003eLatilactobacillus\u003c/em\u003e\u003cem\u003e\u0026nbsp;curvatus\u0026nbsp;\u003c/em\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 33.3333%;\"\u003e\n \u003cp\u003e(n=1; 5.6%)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003eTable 2\u003c/strong\u003e: Identification of \u003cem\u003eLactobacillus\u003c/em\u003e species isolated from chicken gut, tomato, cucumber, and banana using 16S rRNA Gene Sequencing\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"623\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 17.0144%;\"\u003e\n \u003cp\u003e\u003cem\u003eLactobacillus\u003c/em\u003e isolates\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.6693%;\"\u003e\n \u003cp\u003eSource\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 41.252%;\"\u003e\n \u003cp\u003eMolecular identity \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.0642%;\"\u003e\n \u003cp\u003eSimilarity index\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 17.0144%;\"\u003e\n \u003cp\u003eMcya \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.6693%;\"\u003e\n \u003cp\u003eChicken gut \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 41.252%;\"\u003e\n \u003cp\u003e\u003cem\u003eLactobacillus johnsonii \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.0642%;\"\u003e\n \u003cp\u003e100%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 17.0144%;\"\u003e\n \u003cp\u003eMcyb \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.6693%;\"\u003e\n \u003cp\u003eChicken gut \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 41.252%;\"\u003e\n \u003cp\u003e\u003cem\u003eLactobacillus johnsonii \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.0642%;\"\u003e\n \u003cp\u003e100%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 17.0144%;\"\u003e\n \u003cp\u003eMcyc \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.6693%;\"\u003e\n \u003cp\u003eChicken gut \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 41.252%;\"\u003e\n \u003cp\u003e\u003cem\u003eLimosilactobacillus\u003c/em\u003e\u003cem\u003e\u0026nbsp;reuteri\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.0642%;\"\u003e\n \u003cp\u003e100%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 17.0144%;\"\u003e\n \u003cp\u003eMcxb\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.6693%;\"\u003e\n \u003cp\u003eChicken gut \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 41.252%;\"\u003e\n \u003cp\u003e\u003cem\u003eLactobacillus johnsonii \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.0642%;\"\u003e\n \u003cp\u003e100%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 17.0144%;\"\u003e\n \u003cp\u003eMcxc\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.6693%;\"\u003e\n \u003cp\u003eChicken gut \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 41.252%;\"\u003e\n \u003cp\u003e\u003cem\u003eLactobacillus johnsonii \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.0642%;\"\u003e\n \u003cp\u003e100%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 17.0144%;\"\u003e\n \u003cp\u003eChcg\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.6693%;\"\u003e\n \u003cp\u003eChicken gut \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 41.252%;\"\u003e\n \u003cp\u003e\u003cem\u003eLatilactobacillus curvatus\u0026nbsp;\u003c/em\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.0642%;\"\u003e\n \u003cp\u003e100%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 17.0144%;\"\u003e\n \u003cp\u003eChcb\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.6693%;\"\u003e\n \u003cp\u003eChicken gut \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 41.252%;\"\u003e\n \u003cp\u003e\u003cem\u003eLatilactobacillus curvatus\u0026nbsp;\u003c/em\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.0642%;\"\u003e\n \u003cp\u003e100%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 17.0144%;\"\u003e\n \u003cp\u003eChcx2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.6693%;\"\u003e\n \u003cp\u003eChicken gut \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 41.252%;\"\u003e\n \u003cp\u003e\u003cem\u003eLactobacillus johnsonii \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.0642%;\"\u003e\n \u003cp\u003e99.74%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 17.0144%;\"\u003e\n \u003cp\u003eChcx3a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.6693%;\"\u003e\n \u003cp\u003eChicken gut \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 41.252%;\"\u003e\n \u003cp\u003e\u003cem\u003eLimosilactobacillus\u003c/em\u003e\u003cem\u003e\u0026nbsp;reuteri\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.0642%;\"\u003e\n \u003cp\u003e99.47%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 17.0144%;\"\u003e\n \u003cp\u003eChcx3b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.6693%;\"\u003e\n \u003cp\u003eChicken gut \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 41.252%;\"\u003e\n \u003cp\u003e\u003cem\u003eLactobacillus johnsonii \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.0642%;\"\u003e\n \u003cp\u003e100%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 17.0144%;\"\u003e\n \u003cp\u003eChcx4a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.6693%;\"\u003e\n \u003cp\u003eChicken gut \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 41.252%;\"\u003e\n \u003cp\u003e\u003cem\u003eLactobacillus johnsonii \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.0642%;\"\u003e\n \u003cp\u003e100%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 17.0144%;\"\u003e\n \u003cp\u003eMcg \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.6693%;\"\u003e\n \u003cp\u003eChicken gut \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 41.252%;\"\u003e\n \u003cp\u003e\u003cem\u003eLatilactobacillus sakei \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.0642%;\"\u003e\n \u003cp\u003e100%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 17.0144%;\"\u003e\n \u003cp\u003eTo3a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.6693%;\"\u003e\n \u003cp\u003eTomato\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 41.252%;\"\u003e\n \u003cp\u003e\u003cem\u003eLactiplantibacillus plantarum \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.0642%;\"\u003e\n \u003cp\u003e99.54%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 17.0144%;\"\u003e\n \u003cp\u003eTo3b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.6693%;\"\u003e\n \u003cp\u003eTomato\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 41.252%;\"\u003e\n \u003cp\u003e\u003cem\u003eLactiplantibacillus plantarum \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.0642%;\"\u003e\n \u003cp\u003e100%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 17.0144%;\"\u003e\n \u003cp\u003eTo3d\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.6693%;\"\u003e\n \u003cp\u003eTomato\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 41.252%;\"\u003e\n \u003cp\u003e\u003cem\u003eLactiplantibacillus plantarum \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.0642%;\"\u003e\n \u003cp\u003e100%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 17.0144%;\"\u003e\n \u003cp\u003eCu2f\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.6693%;\"\u003e\n \u003cp\u003eCucumber \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 41.252%;\"\u003e\n \u003cp\u003e\u003cem\u003eLatilactobacillus curvatus\u0026nbsp;\u003c/em\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.0642%;\"\u003e\n \u003cp\u003e99.20%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 17.0144%;\"\u003e\n \u003cp\u003eCu3e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.6693%;\"\u003e\n \u003cp\u003eCucumber \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 41.252%;\"\u003e\n \u003cp\u003e\u003cem\u003eLactiplantibacillus plantarum \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.0642%;\"\u003e\n \u003cp\u003e99.73%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 17.0144%;\"\u003e\n \u003cp\u003ePb2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.6693%;\"\u003e\n \u003cp\u003eBanana \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 41.252%;\"\u003e\n \u003cp\u003e\u003cem\u003eLatilactobacillus curvatus\u0026nbsp;\u003c/em\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.0642%;\"\u003e\n \u003cp\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e99.74%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003eTable 3:\u003c/strong\u003e The acid tolerance level of\u003cem\u003e\u0026nbsp;Lactobacillus\u003c/em\u003e isolates after exposure to pH 2.5 and pH 3.0 \u0026nbsp; for 3h\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"432\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 25%;\"\u003e\n \u003cp\u003eIsolates code\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37.5%;\"\u003e\n \u003cp\u003eCell survival rate (%)\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;at pH 2.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37.5%;\"\u003e\n \u003cp\u003eCell survival rate (%)\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;at pH 3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 25%;\"\u003e\n \u003cp\u003e\u003csup\u003e\u0026yen;\u003c/sup\u003eTo3a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37.5%;\"\u003e\n \u003cp\u003e78.39 \u0026nbsp;0.69\u003csup\u003eef\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37.5%;\"\u003e\n \u003cp\u003e97.20 \u0026nbsp;5.79\u003csup\u003eg\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 25%;\"\u003e\n \u003cp\u003eTo3b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37.5%;\"\u003e\n \u003cp\u003e67.82 \u0026nbsp;6.03\u003csup\u003ecd\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37.5%;\"\u003e\n \u003cp\u003e80.74 \u0026nbsp;7.81\u003csup\u003ede\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 25%;\"\u003e\n \u003cp\u003eTo3d\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37.5%;\"\u003e\n \u003cp\u003e73.85 \u0026nbsp;0.84\u003csup\u003ede\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37.5%;\"\u003e\n \u003cp\u003e84.00 \u0026nbsp;1.57\u003csup\u003eef\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 25%;\"\u003e\n \u003cp\u003eChx3a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37.5%;\"\u003e\n \u003cp\u003e83.83 \u0026nbsp;6.06\u003csup\u003efg\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37.5%;\"\u003e\n \u003cp\u003e89.87 \u0026nbsp;1.88\u003csup\u003eefg\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 25%;\"\u003e\n \u003cp\u003eChx3b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37.5%;\"\u003e\n \u003cp\u003e64.46 \u0026nbsp;0.92\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37.5%;\"\u003e\n \u003cp\u003e71.17 \u0026nbsp;7.24\u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 25%;\"\u003e\n \u003cp\u003eCu2f\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37.5%;\"\u003e\n \u003cp\u003e44.99 \u0026nbsp;1.55\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37.5%;\"\u003e\n \u003cp\u003e53.79 \u0026nbsp;2.19\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 25%;\"\u003e\n \u003cp\u003e\u003csup\u003e\u0026yen;\u003c/sup\u003eCu3e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37.5%;\"\u003e\n \u003cp\u003e68.59 \u0026nbsp;3.56\u003csup\u003ecd\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37.5%;\"\u003e\n \u003cp\u003e89.9 \u0026nbsp;3.39\u003csup\u003efg\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 25%;\"\u003e\n \u003cp\u003e\u003csup\u003e\u0026yen;\u003c/sup\u003ePb2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37.5%;\"\u003e\n \u003cp\u003e72.39 \u0026nbsp;6.25\u003csup\u003ede\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37.5%;\"\u003e\n \u003cp\u003e89.57 \u0026nbsp;7.47\u003csup\u003efg\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 25%;\"\u003e\n \u003cp\u003eMcyc\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37.5%;\"\u003e\n \u003cp\u003e89.94 \u0026nbsp;6.68\u003csup\u003eg\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37.5%;\"\u003e\n \u003cp\u003e98.1 \u0026nbsp;1.32\u003csup\u003eg\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 25%;\"\u003e\n \u003cp\u003eMcxb\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37.5%;\"\u003e\n \u003cp\u003e53.35 \u0026nbsp;1.75\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37.5%;\"\u003e\n \u003cp\u003e65.65 \u0026nbsp;2.48\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 25%;\"\u003e\n \u003cp\u003eMcxc\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37.5%;\"\u003e\n \u003cp\u003e63.84 \u0026nbsp;6.12\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37.5%;\"\u003e\n \u003cp\u003e75.71 \u0026nbsp;0.75\u003csup\u003ecd\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 25%;\"\u003e\n \u003cp\u003e\u003csup\u003e\u0026yen;\u003c/sup\u003eGG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37.5%;\"\u003e\n \u003cp\u003e67.65 \u0026nbsp;1.14\u003csup\u003ecd\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37.5%;\"\u003e\n \u003cp\u003e93.03 \u0026nbsp;5.37\u003csup\u003eg\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eThe data shows the mean \u0026plusmn; standard deviation of triplicate values of independent analyzes.\u003c/p\u003e\n\u003cp\u003eColumns with different superscript letters indicate significant differences.\u003c/p\u003e\n\u003cp\u003e\u0026yen; = \u003cem\u003eLactobacillus\u003c/em\u003e isolates for which the survival rate was significantly different and higher at pH 2.5 and 3.0\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 4:\u003c/strong\u003e The bile salt tolerance level of \u003cem\u003eLactobacillus\u003c/em\u003e isolates after exposure to 0.3% and 0.5% bile salt for 4 h\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 24.6575%;\"\u003e\n \u003cp\u003eIsolates code\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38.3562%;\"\u003e\n \u003cp\u003eCell survival rate (%) at 0.3% bile salt\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 36.9863%;\"\u003e\n \u003cp\u003eCell survival rate (%) at 0.5% bile salt\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 24.6575%;\"\u003e\n \u003cp\u003e\u003csup\u003e\u0026pound;\u003c/sup\u003eTo3a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38.3562%;\"\u003e\n \u003cp\u003e98.72 \u0026nbsp;1.37\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 36.9863%;\"\u003e\n \u003cp\u003e89.00 \u0026nbsp;0.75\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 24.6575%;\"\u003e\n \u003cp\u003eTo3b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38.3562%;\"\u003e\n \u003cp\u003e98.74 \u0026nbsp;0.78\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 36.9863%;\"\u003e\n \u003cp\u003e97.42 \u0026nbsp;1.63\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 24.6575%;\"\u003e\n \u003cp\u003eTo3d\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38.3562%;\"\u003e\n \u003cp\u003e98.22 \u0026nbsp;1.60\u003csup\u003ede\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 36.9863%;\"\u003e\n \u003cp\u003e94.57 \u0026nbsp;4.03\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 24.6575%;\"\u003e\n \u003cp\u003eChx3a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38.3562%;\"\u003e\n \u003cp\u003e89.66 \u0026nbsp;5.93\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 36.9863%;\"\u003e\n \u003cp\u003e73.64 \u0026nbsp;1.97\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 24.6575%;\"\u003e\n \u003cp\u003eChx3b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38.3562%;\"\u003e\n \u003cp\u003e81.12 \u0026nbsp;0.85\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 36.9863%;\"\u003e\n \u003cp\u003e62.05 \u0026nbsp;1.51\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 24.6575%;\"\u003e\n \u003cp\u003e\u003csup\u003e\u0026pound;\u003c/sup\u003eCu2f\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38.3562%;\"\u003e\n \u003cp\u003e99.08 \u0026nbsp;1.79\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 36.9863%;\"\u003e\n \u003cp\u003e81.88 \u0026nbsp;1.52\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 24.6575%;\"\u003e\n \u003cp\u003eCu3e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38.3562%;\"\u003e\n \u003cp\u003e95.37 \u0026nbsp;3.80\u003csup\u003ecde\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 36.9863%;\"\u003e\n \u003cp\u003e83.20 \u0026nbsp;6.30\u003csup\u003ecd\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 24.6575%;\"\u003e\n \u003cp\u003ePb2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38.3562%;\"\u003e\n \u003cp\u003e90.87 \u0026nbsp;1.58\u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 36.9863%;\"\u003e\n \u003cp\u003e86.70 \u0026nbsp;1.31\u003csup\u003ecd\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 24.6575%;\"\u003e\n \u003cp\u003eMcyc\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38.3562%;\"\u003e\n \u003cp\u003e98.37 \u0026nbsp;0.86\u003csup\u003ede\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 36.9863%;\"\u003e\n \u003cp\u003e85.25 \u0026nbsp;0.67\u003csup\u003ecd\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 24.6575%;\"\u003e\n \u003cp\u003eMcxb\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38.3562%;\"\u003e\n \u003cp\u003e99.31 \u0026nbsp;0.93\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 36.9863%;\"\u003e\n \u003cp\u003e95.43 \u0026nbsp;2.29\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 24.6575%;\"\u003e\n \u003cp\u003eMcxc\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38.3562%;\"\u003e\n \u003cp\u003e93.38 \u0026nbsp;5.66\u003csup\u003ebcd\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 36.9863%;\"\u003e\n \u003cp\u003e69.64 \u0026nbsp;5.91\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 24.6575%;\"\u003e\n \u003cp\u003e\u003csup\u003e\u0026pound;\u003c/sup\u003eGG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38.3562%;\"\u003e\n \u003cp\u003e92.92 \u0026nbsp;0.97\u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 36.9863%;\"\u003e\n \u003cp\u003e88.77 \u0026nbsp;3.08\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eThe data shows mean \u0026plusmn; standard deviation of triplicate values of independent analyzes.\u003c/p\u003e\n\u003cp\u003eColumns with different superscript letters indicate significant differences.\u003c/p\u003e\n\u003cp\u003e\u0026pound; = \u003cem\u003eLactobacillus\u0026nbsp;\u003c/em\u003eisolates for which the survival rate was significantly different and higher at 0.3% and 0.5% bile salt\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 5:\u0026nbsp;\u003c/strong\u003eThe BSH activity, EPS production ability, and cholesterol-lowering ability of \u003cem\u003eLactobacillus\u0026nbsp;\u003c/em\u003eisolates\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"486\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 17.4897%;\"\u003e\n \u003cp\u003eIsolates code\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 22.0165%;\"\u003e\n \u003cp\u003eBSH activity\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 28.3951%;\"\u003e\n \u003cp\u003eEPS producing ability\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 32.0988%;\"\u003e\n \u003cp\u003eCholesterol-lowering ability\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 17.4897%;\"\u003e\n \u003cp\u003eTo3a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 22.0165%;\"\u003e\n \u003cp\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 28.3951%;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 32.0988%;\"\u003e\n \u003cp\u003e49 \u0026nbsp;1.27\u003csup\u003eh\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 17.4897%;\"\u003e\n \u003cp\u003eTo3b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 22.0165%;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 28.3951%;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 32.0988%;\"\u003e\n \u003cp\u003e40 \u0026nbsp;1.17\u003csup\u003eef\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 17.4897%;\"\u003e\n \u003cp\u003eTo3d\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 22.0165%;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 28.3951%;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 32.0988%;\"\u003e\n \u003cp\u003e\u0026nbsp; 0.20\u003csup\u003ei\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 17.4897%;\"\u003e\n \u003cp\u003eChx3a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 22.0165%;\"\u003e\n \u003cp\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 28.3951%;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 32.0988%;\"\u003e\n \u003cp\u003e\u0026nbsp; 1.37\u003csup\u003ej\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 17.4897%;\"\u003e\n \u003cp\u003eChx3b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 22.0165%;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 28.3951%;\"\u003e\n \u003cp\u003e\u0026ndash;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 32.0988%;\"\u003e\n \u003cp\u003e\u0026nbsp; 0.19\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 17.4897%;\"\u003e\n \u003cp\u003eCu2f\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 22.0165%;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 28.3951%;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 32.0988%;\"\u003e\n \u003cp\u003e\u0026nbsp; 1.58\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 17.4897%;\"\u003e\n \u003cp\u003eCu3e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 22.0165%;\"\u003e\n \u003cp\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 28.3951%;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 32.0988%;\"\u003e\n \u003cp\u003e46 \u0026nbsp;1.73\u003csup\u003eg\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 17.4897%;\"\u003e\n \u003cp\u003ePb2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 22.0165%;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 28.3951%;\"\u003e\n \u003cp\u003e\u0026ndash;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 32.0988%;\"\u003e\n \u003cp\u003e\u0026nbsp; 0.98\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 17.4897%;\"\u003e\n \u003cp\u003eMcyc\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 22.0165%;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 28.3951%;\"\u003e\n \u003cp\u003e\u0026ndash;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 32.0988%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 17.4897%;\"\u003e\n \u003cp\u003eMcxb\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 22.0165%;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 28.3951%;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 32.0988%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 17.4897%;\"\u003e\n \u003cp\u003eMcxc\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 22.0165%;\"\u003e\n \u003cp\u003e\u0026ndash;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 28.3951%;\"\u003e\n \u003cp\u003e\u0026ndash;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 32.0988%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 17.4897%;\"\u003e\n \u003cp\u003eGG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 22.0165%;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 28.3951%;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 32.0988%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003ef\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003csup\u003ea\u003c/sup\u003eBSH activity is shown based on the diameter of precipitation zones: No precipitation (\u0026ndash;), precipitation zone (+) up to 10 mm, precipitation zone 10\u0026ndash;15 mm (++), and precipitation zone \u0026gt;15 mm (+++).\u003c/p\u003e\n\u003cp\u003e\u003csup\u003eb\u003c/sup\u003eCholesterol\u0026ndash;lowering ability values are mean \u0026plusmn; SD of two independent observations (n=2) of each sample.\u003c/p\u003e\n\u003cp\u003e\u003csup\u003eabcd\u003c/sup\u003eMeans bearing different superscripts in a column differ significantly (P\u0026lt;0.05).\u003c/p\u003e\n\u003cp\u003e\u003csup\u003ec\u003c/sup\u003eEPS producing ability is displayed based on\u003csup\u003e\u0026nbsp;\u003c/sup\u003e(+) means positive growth while (\u0026ndash;) means negative growth\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 6:\u003c/strong\u003e DPPH scavenging activity (%) of \u003cem\u003eLactobacillus\u003c/em\u003e isolates\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 48.3173%;\"\u003e\n \u003cp\u003eAntioxidant assay\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 51.6827%;\"\u003e\n \u003cp\u003eDPPH scavenging activity (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 48.3173%;\"\u003e\n \u003cp\u003eTo3a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 51.6827%;\"\u003e\n \u003cp\u003e19.73 \u0026plusmn;2.28\u003csup\u003ecd\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 48.3173%;\"\u003e\n \u003cp\u003eTo3b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 51.6827%;\"\u003e\n \u003cp\u003e17.92\u0026plusmn;1.09\u003csup\u003ecd\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 48.3173%;\"\u003e\n \u003cp\u003e\u003csup\u003e\u0026euro;\u003c/sup\u003eTo3d\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 51.6827%;\"\u003e\n \u003cp\u003e24.45\u0026plusmn;1.15\u003csup\u003ef\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 48.3173%;\"\u003e\n \u003cp\u003eChcx3a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 51.6827%;\"\u003e\n \u003cp\u003e18.26\u0026plusmn;0.63\u003csup\u003ecd\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 48.3173%;\"\u003e\n \u003cp\u003eChcx3b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 51.6827%;\"\u003e\n \u003cp\u003e17.08\u0026plusmn;1.65\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 48.3173%;\"\u003e\n \u003cp\u003eCu2f\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 51.6827%;\"\u003e\n \u003cp\u003e6.81\u0026plusmn;1.56\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 48.3173%;\"\u003e\n \u003cp\u003eCu3e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 51.6827%;\"\u003e\n \u003cp\u003e18.05\u0026plusmn;2.93\u003csup\u003ecd\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 48.3173%;\"\u003e\n \u003cp\u003ePb2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 51.6827%;\"\u003e\n \u003cp\u003e11.11\u0026plusmn;2.25\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 48.3173%;\"\u003e\n \u003cp\u003e\u003csup\u003e#\u003c/sup\u003eMcyc\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 51.6827%;\"\u003e\n \u003cp\u003e20.90\u0026plusmn;1.62\u003csup\u003ede\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 48.3173%;\"\u003e\n \u003cp\u003eMcxb\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 51.6827%;\"\u003e\n \u003cp\u003e7.85\u0026plusmn;1.36\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 48.3173%;\"\u003e\n \u003cp\u003eMcxc\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 51.6827%;\"\u003e\n \u003cp\u003e13.33\u0026plusmn;1.66\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 48.3173%;\"\u003e\n \u003cp\u003eGG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 51.6827%;\"\u003e\n \u003cp\u003e23.20\u0026plusmn;1.35\u003csup\u003eef\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 48.3173%;\"\u003e\n \u003cp\u003eAscorbic acid (10mg/ml)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 51.6827%;\"\u003e\n \u003cp\u003e35.63\u0026plusmn;2.20\u003csup\u003eg\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eValues are mean \u0026plusmn; SD of three independent observations (n=3) of each sample.\u003c/p\u003e\n\u003cp\u003e\u003csup\u003eabcd\u003c/sup\u003eMeans bearing different superscripts in a column differ significantly (P\u0026lt;0.05)\u003c/p\u003e\n\u003cp\u003e\u0026euro; = \u003cem\u003eLactobacillus\u003c/em\u003e isolates for which the DPPH scavenging activity was significantly (p\u0026le;0.05) higher than that of the reference strain\u003c/p\u003e\n\u003cp\u003e# = \u003cem\u003eLactobacillus\u0026nbsp;\u003c/em\u003eisolates for which the DPPH scavenging activity was not significantly different (p\u0026le;0.05) from that of the reference strain\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 7:\u003c/strong\u003e Antimicrobial activity of CFS of \u003cem\u003eLactobacillus\u003c/em\u003e against pathogenic organisms\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"504\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" style=\"width: 101px;\"\u003e\n \u003cp\u003eIsolates code\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" valign=\"top\" style=\"width: 403px;\"\u003e\n \u003cp\u003eZone of inhibition\u003csup\u003ea\u003c/sup\u003e (mm)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 97px;\"\u003e\n \u003cp\u003e\u003cem\u003eE\u003c/em\u003e\u003cem\u003e. coli\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e\u003cem\u003eS.\u003c/em\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003cem\u003eaureus\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 81px;\"\u003e\n \u003cp\u003e\u003cem\u003eB\u003c/em\u003e\u003cem\u003e. cereus\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 129px;\"\u003e\n \u003cp\u003e\u003cem\u003eL\u003c/em\u003e\u003cem\u003e. monocytogenes\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 101px;\"\u003e\n \u003cp\u003e\u003csup\u003e\u0026infin;\u003c/sup\u003eTo3a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 97px;\"\u003e\n \u003cp\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 81px;\"\u003e\n \u003cp\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 129px;\"\u003e\n \u003cp\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 101px;\"\u003e\n \u003cp\u003e\u003csup\u003e\u0026infin;\u003c/sup\u003eTo3b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 97px;\"\u003e\n \u003cp\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 81px;\"\u003e\n \u003cp\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 129px;\"\u003e\n \u003cp\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 101px;\"\u003e\n \u003cp\u003e\u003csup\u003e\u0026infin;\u003c/sup\u003eTo3d\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 97px;\"\u003e\n \u003cp\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 81px;\"\u003e\n \u003cp\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 129px;\"\u003e\n \u003cp\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 101px;\"\u003e\n \u003cp\u003eChx3a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 97px;\"\u003e\n \u003cp\u003e\u0026ndash;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e\u0026ndash;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 81px;\"\u003e\n \u003cp\u003e\u0026ndash;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 129px;\"\u003e\n \u003cp\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 101px;\"\u003e\n \u003cp\u003eChx3b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 97px;\"\u003e\n \u003cp\u003e\u0026ndash;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e\u0026ndash;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 81px;\"\u003e\n \u003cp\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 129px;\"\u003e\n \u003cp\u003e\u0026ndash;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 101px;\"\u003e\n \u003cp\u003eCu2f\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 97px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 81px;\"\u003e\n \u003cp\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 129px;\"\u003e\n \u003cp\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 101px;\"\u003e\n \u003cp\u003eCu3e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 97px;\"\u003e\n \u003cp\u003e\u0026ndash;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e\u0026ndash;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 81px;\"\u003e\n \u003cp\u003e\u0026ndash;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 129px;\"\u003e\n \u003cp\u003e\u0026ndash;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 101px;\"\u003e\n \u003cp\u003ePb2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 97px;\"\u003e\n \u003cp\u003e\u0026ndash;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e\u0026ndash;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 81px;\"\u003e\n \u003cp\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 129px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 101px;\"\u003e\n \u003cp\u003eMcyc\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 97px;\"\u003e\n \u003cp\u003e\u0026ndash;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e\u0026ndash;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 81px;\"\u003e\n \u003cp\u003e\u0026ndash;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 129px;\"\u003e\n \u003cp\u003e\u0026ndash;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 101px;\"\u003e\n \u003cp\u003eMcxb\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 97px;\"\u003e\n \u003cp\u003e\u0026ndash;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e\u0026ndash;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 81px;\"\u003e\n \u003cp\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 129px;\"\u003e\n \u003cp\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 101px;\"\u003e\n \u003cp\u003eMcxc\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 97px;\"\u003e\n \u003cp\u003e\u0026ndash;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e\u0026ndash;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 81px;\"\u003e\n \u003cp\u003e\u0026ndash;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 129px;\"\u003e\n \u003cp\u003e\u0026ndash;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 101px;\"\u003e\n \u003cp\u003eGG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 97px;\"\u003e\n \u003cp\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 81px;\"\u003e\n \u003cp\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 129px;\"\u003e\n \u003cp\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003csup\u003ea\u003c/sup\u003eResults of independent experiments (n =3) of inhibition zones: (\u0026ndash;) = not detected; (+) = inhibitory zone \u0026lt; 10mm; (++) = inhibitory zone (10\u0026ndash;15 mm), and (+++) = inhibitory zone \u0026gt; 15 mm.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026infin; = \u003cem\u003eLactobacillus\u003c/em\u003e isolates with antimicrobial activity similar with that of the reference strain (GG), with an inhibition zone of 10 to 15 mm or more against \u003cem\u003eE. coli\u003c/em\u003e, \u003cem\u003eS. aureus\u003c/em\u003e, \u003cem\u003eB. cereus\u003c/em\u003e and \u003cem\u003eL. monocytogenes\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 8:\u0026nbsp;\u003c/strong\u003eThe\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eauto-aggregation ability of \u003cem\u003eLactobacillus\u003c/em\u003e isolates after 2 h and 4 h incubation\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" style=\"width: 187px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003eIsolates code\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 434px;\"\u003e\n \u003cp\u003eAuto-aggregation (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 217px;\"\u003e\n \u003cp\u003e2h\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 217px;\"\u003e\n \u003cp\u003e4h\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 187px;\"\u003e\n \u003cp\u003e\u003csup\u003e\u0026micro;\u003c/sup\u003eTo3a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 217px;\"\u003e\n \u003cp\u003e24\u0026nbsp;4.00\u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 217px;\"\u003e\n \u003cp\u003e\u0026nbsp;4.62\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 187px;\"\u003e\n \u003cp\u003eTo3b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 217px;\"\u003e\n \u003cp\u003e22. \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 217px;\"\u003e\n \u003cp\u003e29.33 \u003csup\u003ebcd\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 187px;\"\u003e\n \u003cp\u003e\u003csup\u003e\u0026micro;\u003c/sup\u003eTo3d\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 217px;\"\u003e\n \u003cp\u003e26.67 \u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 217px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003ecd\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 187px;\"\u003e\n \u003cp\u003e\u003csup\u003e\u0026micro;\u003c/sup\u003eChcx3a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 217px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 217px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003ecd\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 187px;\"\u003e\n \u003cp\u003e\u003csup\u003e\u0026micro;\u003c/sup\u003eChcx3b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 217px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 217px;\"\u003e\n \u003cp\u003e48.67 \u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 187px;\"\u003e\n \u003cp\u003e\u003csup\u003e\u0026micro;\u003c/sup\u003eCu2f\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 217px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 217px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 187px;\"\u003e\n \u003cp\u003eCu3e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 217px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 217px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003eabc\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 187px;\"\u003e\n \u003cp\u003ePb2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 217px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 217px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 187px;\"\u003e\n \u003cp\u003e\u003csup\u003e\u0026micro;\u003c/sup\u003eMcyc\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 217px;\"\u003e\n \u003cp\u003e33.33 \u003csup\u003ecd\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 217px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 187px;\"\u003e\n \u003cp\u003eMcxb\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 217px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 217px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 187px;\"\u003e\n \u003cp\u003eMcxc\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 217px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 217px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003ecd\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 187px;\"\u003e\n \u003cp\u003eGG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 217px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 217px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003eabcd\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eValues are mean \u0026plusmn; SD of three independent observations (n=3) of each sample.\u003c/p\u003e\n\u003cp\u003e\u003csup\u003eabcd\u0026nbsp;\u003c/sup\u003eMeans bearing different superscripts in a column differ significantly (P\u0026lt;0.05).\u003c/p\u003e\n\u003cp\u003e\u0026micro; = \u003cem\u003eLactobacillus\u003c/em\u003e isolates with a significant (p \u0026le; 0.05) increase in auto-aggregation (%) from 2 to 4 h incubation and whose auto-aggregation (%) was significantly higher than that of the reference strain after 2 and 3 h incubation, respectively\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 9:\u0026nbsp;\u003c/strong\u003eCell surface hydrophobicity ability of \u003cem\u003eLactobacillus\u003c/em\u003e isolates after 1 hr incubation\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 227px;\"\u003e\n \u003cp\u003eIsolates\u0026nbsp;code\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 264px;\"\u003e\n \u003cp\u003eCell surface hydrophobicity (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 227px;\"\u003e\n \u003cp\u003eTo3a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 264px;\"\u003e\n \u003cp\u003e16.00 \u0026nbsp;4.00\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 227px;\"\u003e\n \u003cp\u003eTo3b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 264px;\"\u003e\n \u003cp\u003e18.67 \u003csup\u003ebcd\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 227px;\"\u003e\n \u003cp\u003eTo3d\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 264px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 227px;\"\u003e\n \u003cp\u003eChcx3a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 264px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 227px;\"\u003e\n \u003cp\u003e\u003csup\u003e#\u003c/sup\u003eChcx3b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 264px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 227px;\"\u003e\n \u003cp\u003e\u003csup\u003e\u0026euro;\u003c/sup\u003eCu2f\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 264px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003eg\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 227px;\"\u003e\n \u003cp\u003e\u003csup\u003e\u0026euro;\u003c/sup\u003eCu3e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 264px;\"\u003e\n \u003cp\u003e72.00 \u003csup\u003ef\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 227px;\"\u003e\n \u003cp\u003ePb2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 264px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 227px;\"\u003e\n \u003cp\u003e\u003csup\u003e\u0026euro;\u003c/sup\u003eMcyc\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 264px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003eg\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 227px;\"\u003e\n \u003cp\u003eMcxb\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 264px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 227px;\"\u003e\n \u003cp\u003eMcxc\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 264px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003ecd\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 227px;\"\u003e\n \u003cp\u003eGG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 264px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eValues are mean \u0026plusmn; SD of three independent observations (n=3) each sample.\u003c/p\u003e\n\u003cp\u003e\u003csup\u003eabcd\u0026nbsp;\u003c/sup\u003eMeans bearing different superscripts in a column differ significantly (P\u0026lt;0.05).\u003c/p\u003e\n\u003cp\u003e\u0026euro; = \u003cem\u003eLactobacillus\u003c/em\u003e isolates for which the cell surface hydrophobicity values were significantly (p\u0026le;0.05) higher than those of the reference strain.\u003c/p\u003e\n\u003cp\u003e# = \u003cem\u003eLactobacillus\u003c/em\u003e isolates for which the cell surface hydrophobicity values were not significantly different (p\u0026gt;0.05) from that of the reference strain\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 10:\u003c/strong\u003e Antibiotic susceptibility profile of \u003cem\u003eLactobacillus\u003c/em\u003e isolates using the disk diffusion method\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"653\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003eAntibiotics\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"13\" valign=\"top\" style=\"width: 549px;\"\u003e\n \u003cp\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; Susceptibility profile of \u003cem\u003eLactobacillus i\u003c/em\u003esolates\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eTo3a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eTo3b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eTo3d\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eChx3a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eChx3b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eCu2f\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eCu3e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003ePb2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eMcyc\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eMcxb\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eMcxc\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003eGG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 1px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003ePenicillin G (10 \u0026mu;g)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 1px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003eChloramphenicol (30 \u0026mu;g)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 1px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003eKanamycin (30 \u0026mu;g)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 1px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003eGentamycin (10 \u0026mu;g)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 1px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003eStreptomycin (10 \u0026mu;g)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 1px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003eTetracycline (30 \u0026mu;g)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eI\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 1px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003eErythromycin (15 \u0026mu;g)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 1px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003eAmpicillin (10 \u0026mu;g)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 1px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003eVancomycin (30 \u0026mu;g)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eI\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 1px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eResults are presented as S: Susceptible (\u0026ge;21 mm), I: intermediate (16\u0026ndash;20 mm), R: resistance (\u0026le; 15 mm)\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 11:\u0026nbsp;\u003c/strong\u003eBlood\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003ehaemolysis and DNase activity of \u003cem\u003eLactobacillus\u003c/em\u003e isolates\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 108px;\"\u003e\n \u003cp\u003eIsolates code\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 102px;\"\u003e\n \u003cp\u003eHaemolysis\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003eDNase activity\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 108px;\"\u003e\n \u003cp\u003eTo3a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 102px;\"\u003e\n \u003cp\u003e\u0026upsih;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e\u0026ndash;ve\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 108px;\"\u003e\n \u003cp\u003eTo3b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 102px;\"\u003e\n \u003cp\u003e\u0026upsih;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e\u0026ndash;ve\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 108px;\"\u003e\n \u003cp\u003eTo3d\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 102px;\"\u003e\n \u003cp\u003e\u0026upsih;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e\u0026ndash;ve\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 108px;\"\u003e\n \u003cp\u003eChx3a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 102px;\"\u003e\n \u003cp\u003e\u0026upsih;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e\u0026ndash;ve\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 108px;\"\u003e\n \u003cp\u003eChx3b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 102px;\"\u003e\n \u003cp\u003e\u0026upsih;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e\u0026ndash;ve\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 108px;\"\u003e\n \u003cp\u003eCu2f\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 102px;\"\u003e\n \u003cp\u003e\u0026upsih;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e\u0026ndash;ve\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 108px;\"\u003e\n \u003cp\u003eCu3e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 102px;\"\u003e\n \u003cp\u003e\u0026upsih;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e\u0026ndash;ve\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 108px;\"\u003e\n \u003cp\u003ePb2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 102px;\"\u003e\n \u003cp\u003e\u0026upsih;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e\u0026ndash;ve\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 108px;\"\u003e\n \u003cp\u003eMcya\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 102px;\"\u003e\n \u003cp\u003e\u0026upsih;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e\u0026ndash;ve\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 108px;\"\u003e\n \u003cp\u003eMcxb\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 102px;\"\u003e\n \u003cp\u003e\u0026upsih;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e\u0026ndash;ve\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 108px;\"\u003e\n \u003cp\u003eMcxc\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 102px;\"\u003e\n \u003cp\u003e\u0026upsih;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e\u0026ndash;ve\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 108px;\"\u003e\n \u003cp\u003eGG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 102px;\"\u003e\n \u003cp\u003e\u0026upsih;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e\u0026ndash;ve\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003csup\u003ea\u0026nbsp;\u003c/sup\u003e\u0026upsih;: means no haemolysis, \u0026beta;: means complete haemolysis, \u0026alpha;: means partial haemolysis.\u003c/p\u003e\n\u003cp\u003e\u003csup\u003eb\u003c/sup\u003e \u0026ndash;ve indicates no DNase activity, +ve indicates DNase activity\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 12:\u0026nbsp;\u003c/strong\u003eAdhesion capacity of \u003cem\u003eLactobacillus\u003c/em\u003e isolates to Caco-2 epithelial cells\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eIsolates code\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 151px;\"\u003e\n \u003cp\u003eAdhesion (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eTo3a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 151px;\"\u003e\n \u003cp\u003e9.94 \u0026nbsp;.06\u003csup\u003ede\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eTo3b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 151px;\"\u003e\n \u003cp\u003e6.85 \u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eTo3d\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 151px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003eg\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eChcx3a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 151px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eChcx3b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 151px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003eh\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eCu2f\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 151px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eCu3e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 151px;\"\u003e\n \u003cp\u003e3.58 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003ePb2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 151px;\"\u003e\n \u003cp\u003e2.60\u0026nbsp;0.83\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eMcyc\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 151px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003eef\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eMcxb\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 151px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eMcxc\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 151px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 151px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003csup\u003ef\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eValues are mean \u0026plusmn; SD of three independent observations (n=3).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003csup\u003eabcd\u0026nbsp;\u003c/sup\u003eMeans bearing different superscripts in a column differ significantly (P\u0026lt;0.05)\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 13:\u0026nbsp;\u003c/strong\u003eCumulative probiotic potential (CPP) of \u003cem\u003eLactobacillus\u003c/em\u003e isolates\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"735\" class=\"fr-table-selection-hover\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 135px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003eIndicator score\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003eTo3a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003eTo3b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003eTo3d\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003eChcx3a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003eChcx3b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003eCu2f\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003eCu3e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43px;\"\u003e\n \u003cp\u003ePb2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 55px;\"\u003e\n \u003cp\u003eMcyc\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 55px;\"\u003e\n \u003cp\u003eMcxb\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 61px;\"\u003e\n \u003cp\u003eMcxc\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 1px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"14\" valign=\"top\" style=\"width: 735px;\"\u003e\n \u003cp\u003eProbiotic characters\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 135px;\"\u003e\n \u003cp\u003eAcid tolerance\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003e% \u0026ge; 50 = 1\u003c/p\u003e\n \u003cp\u003e% \u0026lt; 50 = 0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 55px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 55px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 61px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 1px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 135px;\"\u003e\n \u003cp\u003eBile salt tolerance\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003e% \u0026ge; 50 = 1\u003c/p\u003e\n \u003cp\u003e% \u0026lt; 50 = 0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 55px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 55px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 61px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 1px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 135px;\"\u003e\n \u003cp\u003eAuto-aggregation \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003e% \u0026le; 30 = 0\u003c/p\u003e\n \u003cp\u003e% \u0026gt; 30 = 1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 55px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 55px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 61px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 1px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 135px;\"\u003e\n \u003cp\u003eCell surface hydrophobicity\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003e% \u0026le; 30 = 0\u003c/p\u003e\n \u003cp\u003e% \u0026gt; 30 = 1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 55px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 55px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 61px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 1px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 135px;\"\u003e\n \u003cp\u003eAntimicrobial activity\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003e\u0026lt; 15 mm = 0\u003c/p\u003e\n \u003cp\u003e\u0026ge; 15 mm = 1\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 55px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 55px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 61px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 1px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"14\" valign=\"top\" style=\"width: 735px;\"\u003e\n \u003cp\u003eSafety parameters\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 135px;\"\u003e\n \u003cp\u003e\u0026nbsp;Tetracycline antibiotic susceptibility\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003eSensitive/ intrinsic resistant=0\u003c/p\u003e\n \u003cp\u003eResistant=1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 55px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 55px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 61px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 1px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 135px;\"\u003e\n \u003cp\u003eHaemolytic activity\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003eActivity=0\u003c/p\u003e\n \u003cp\u003eNo activity=1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 55px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 55px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 61px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 1px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 135px;\"\u003e\n \u003cp\u003eDNase activity\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003eActivity=0\u003c/p\u003e\n \u003cp\u003eNo activity=1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 55px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 55px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 61px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 1px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 239px;\"\u003e\n \u003cp\u003eTotal score\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e7/8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e6/8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e7/8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e5/8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e6/8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e7/8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e5/8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43px;\"\u003e\n \u003cp\u003e4/8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 55px;\"\u003e\n \u003cp\u003e6/8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 55px;\"\u003e\n \u003cp\u003e4/8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 61px;\"\u003e\n \u003cp\u003e5/8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 1px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 239px;\"\u003e\n \u003cp\u003eCumulative probiotic potential\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e87.50%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e75%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e87.50%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e62.\u003c/p\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003cp\u003e%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e75\u003c/p\u003e\n \u003cp\u003e%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e87.50%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e62.\u003c/p\u003e\n \u003cp\u003e50%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43px;\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003cp\u003e%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 55px;\"\u003e\n \u003cp\u003e75\u003c/p\u003e\n \u003cp\u003e%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 55px;\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003cp\u003e%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 61px;\"\u003e\n \u003cp\u003e62.\u003c/p\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003cp\u003e%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 1px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Lactobacillus, probiotic, fruits, vegetables, animal gut, food sources","lastPublishedDoi":"10.21203/rs.3.rs-7594770/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7594770/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground: \u003c/strong\u003eThere has been a growing interest among researchers in identifying highly functional probiotic \u003cem\u003eLactobacillus\u003c/em\u003e species from plant and animal sources in recent times. This study aims to prospect and characterize \u003cem\u003eLactobacillus\u003c/em\u003e species with probiotic potential from animal gut, fruit, and vegetable sources.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethod:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eLactobacillus\u003c/em\u003especies were isolated from samples using selective enrichment techniques. Colony morphology, catalase testing, and Gram staining were used for the preliminary identification of the presumptive \u003cem\u003eLactobacillus\u003c/em\u003e strains. Their identities were subsequently confirmed by 16S rRNA gene sequencing and BLAST analysis. The identified \u003cem\u003eLactobacillus\u003c/em\u003e strains were subjected to in vitro characterization to assess their probiotic properties.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults: \u003c/strong\u003eTen \u003cem\u003eLactobacillus\u003c/em\u003e species isolates demonstrated high acid tolerance, with survivability exceeding 50% at pH 2.5 and 3.0. Their survival rates ranged from 62.05% to 97.42% and from 81.12% to 99.31% in the presence of 0.5% and 0.3% bile salt concentrations, respectively. All the isolates exhibited varying capacities to reduce cholesterol (13.06% –57.64%) and demonstrated antioxidant activity ranging from 6.81% to 24.45%. Only one isolate (Mcxc) lacked bile salt hydrolase activity, and 33.33% of the isolates did not produce any exopolysaccharides. The auto-aggregation abilities ranged from 18% to 52% after 4 h of incubation, while cell surface hydrophobicity ranged from 0.00% to 84.68%. None of the isolates exhibited haemolytic or DNase activity. The degree of adhesion to Caco-2 cells ranged from 2.6% to 15.29%. All isolates were susceptible to erythromycin, chloramphenicol, ampicillin, and penicillin G. Notably, only four \u003cem\u003eLactobacillus\u003c/em\u003e isolates exhibited inhibitory activity against all the tested pathogenic bacteria.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusions: \u003c/strong\u003eOur findings revealed that \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e To3a and To3d, isolated from tomatoes, stand out as a viable and alternative source of probiotics due to their promising probiotic properties and health-promoting benefits; thus, they may be explored for practical use in novel, locally adaptable functional products.\u003c/p\u003e","manuscriptTitle":"Prospecting and characterization of potential probiotic Lactobacillus species from animal gut and food sources","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-10-17 18:52:02","doi":"10.21203/rs.3.rs-7594770/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"63bd7c4e-74fb-4ee9-a744-989ab4487de7","owner":[],"postedDate":"October 17th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-11-05T07:23:43+00:00","versionOfRecord":[],"versionCreatedAt":"2025-10-17 18:52:02","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7594770","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7594770","identity":"rs-7594770","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}