Autochthonous Lactic Acid Bacteria from Kadidd as Functional Starter Culture to Improve quality and safety

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Segli, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5005817/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 23 Apr, 2025 Read the published version in Discover Food → Version 1 posted 12 You are reading this latest preprint version Abstract Lactic acid bacteria (LAB) from meat dry-salted Kaddid were evaluated using spoilage and safety criteria as a first step of multivariable selection. LAB were isolated, identified and characterized from Kaddid prepared under controlled conditions. After preliminary physiological characterization, isolates ( n = 39) were subjected to (GTG)5-PCR analysis and 16S rRNA sequencing. Major LAB populations were, Lactiplantibacillus paraplantarum (41%), Latilactobacillus sakei/ subsp. sakei (25%), Enterococcus faecium (13%), Pediococcus acidilactici (10%), Enterococcus hirae (6%) and Weissella cibaria (5%). Antagonistic activity against pathogens/contaminants and yeast/molds showed strains with antilisterial and antifungal activity. Bacteriocin-like inhibitory substances (BLIS) showed high titles (AU/mL) against L. monocytogenes FBUNT. The examination of bacteriocin genes revealed Lpb. paraplantarum K18, Lat. sakei K25 and Lat. sakei subsp. sakei K22, K26 strains harboring skg A and sak P structural genes encoding for sakacin G and P bacteriocins. Resistance/susceptibility of lactobacilli strains to antibiotics showed high phenotypic resistance to TET while multi-resistance pattern was displayed by enterococci. Genotypic characterization exhibited only tet M out of 15 assayed genes, conferring resistance to TET. As a quality criterion, no exopolysaccharides and biogenic amines production were detected in the evaluated strains. Based on these results, Lpb. paraplantarum K18 and K21 strains, with inhibitory and antifungal activities, phenotypical susceptibility to the assayed antimicrobial compounds and low risk as spoilers, could be used as functional starter cultures for safe Kaddid production. Kaddid fermentation Lpb. paraplantarum Lat. sakei bacteriocins antimicrobial resistance Figures Figure 1 1. Introduction Kaddid or El-Guedid is a traditional meat product prepared all over the Algerian territory from sheep and beef meat whereas goat and camel meat is mainly used in sub-dry areas. Usually, the raw meat is cut into strips, seasoned abundantly with salt and spices and brine or dry salting was applied before dried either under the sun or in the shade [ 1 ]. Southwestern Algerian Kaddid is mostly produced by adding salt and spices onto fresh lamb meat strips that are then hung to dry in the shade [ 2 ]. As part of Kaddid microbiota, LAB were reported to mainly involve species adapted to high NaCl concentration as Lactiplantibacillus (Lpb.) plantarum, Latilactobacillus (Lat.) sakei and Lat. curvatus described for Tunisian and Algerian dry-salted meat and El-Guedid [ 1 , 3 ]. During fermentation, lactobacilli produce organic acids causing a pH reduction, but can also produce exopolysaccharides (EPS) reported to negatively affect the texture of fermented dry and salted meat products [ 4 ]. LAB can also produce biogenic amines by amino acid decarboxylation that accumulates during fermentation/ripening, which influence organoleptic properties and health when consumed in high concentrations [ 5 ]. Lactobacilli have a biopreservative role in fermented meat products by producing, in addition to organic acids, antimicrobial compounds such as bacteriocins. These ribosomal produced peptides present a variable spectrum of antimicrobial activity, usually against related species such as L. monocyotgenes . This bacterium can infect humans and animals as opportunistic foodborne pathogen causing serious illness and is frequently isolated from food and food-processing environments [ 6 ]. Bacteriocinogenic lactobacilli as bioprotective cultures are potentially used as natural shelf-life safety and preventing the growth of foodborne pathogens [ 7 ]. Another relevant safety aspect of LAB is the presence of antibiotic resistance genes in mobile genetic elements, such as plasmids and transposons. In fact, these elements can be transferred to other species, including pathogenic bacteria, during food manufacture or during the passage through the gastrointestinal tract. Indeed, this poses an additional risk due to the ready-to-eat nature of the fermented meat products and their potential to become important antibiotic resistance reservoirs [ 8 ]. Particularly, the presence of tetracycline and erythromycin resistant lactobacilli in meat-associated bacteria was reported [ 9 ]. To avoid the spread of antibiotics resistance genes that may be part of GRAS LAB genome, beneficial LAB to be used as starter cultures must be evaluated on a strain basis [ 10 ]. Therefore, the aim of this study was the isolation, identification and characterization of LAB from laboratory-made Kaddid to select an autochthonous functional culture with a main focus to prevent spoilage and ensure product safety. 2. Material and methods 2.1. Kaddid preparation and sampling A leg lamb meat (~ 3 kg) 24 h post-mortem was purchased from a butcher’s shop in Médéa, Algeria. Kaddid preparation was carried out according to South-Western Algerian method [ 2 ]. Meat was aseptically skinned and deboned manually and thin (2–3 cm thick) and long (20–30 cm) stripes were cut following anatomical lines. The strips were then homogeneously rubbed with a mix of salt (200 g), black pepper (2 g) and cumin (2 g) while sugar (10 g) was added to boost LAB growth and placed in a ventilated oven (Memmert, Germany) at 22 ± 1°C and 75–80% (RH%) for 15 days. Physicochemical and microbiological analyses were carried out during drying-fermentation at 0, 3, 7 and 15 days. 2.2. Physicochemical and microbiological characterization of Kaddid. Isolation of LAB Total titratable acidity (TTA), pH, moisture (%RH), water activity (a W ) and microbiological analysis of Kaddid samples were performed as previously described Boubakri et al. [ 2 ]. For microbiological analysis, each sample (25 g) suspended in 225 mL of sterile tryptone-salt (tryptone 1 g/L; NaCl 0.85 g/L; Tween 80 1 mL/L) and homogenized (Stomacher 400, Seward, Worthing, UK) was serially diluted. Viable cells enumeration of total aerobic mesophilic bacteria (TAMB) on Plate Count Agar (Britania, Argentina) incubated aerobically (48 h, 30°C) was carried out. In addition, counts of total coliforms, staphylococci and the detection of Salmonella were achieved as described by Boubakri et al. [ 2 ]. LAB viable counts were performed using MRS agar (Merck, Germany) microaerobically incubated at 28°C for 4 days. LAB isolation was carried out after 15 days of Kaddid ripening by randomly picking forty (40) single colonies from MRS plates and after purification by successive streaking, Gram and catalase tests were performed. 2.3. Identification of lactic acid bacteria LAB were preliminary grouped by their physiological and biochemical traits as previously described Boubakri et al. [ 11 ]. Growth at 10 ºC and 45 ºC during 7 d and 24 h, respectively, at pH 4 to 9.6 and in the presence of 4 to 10% NaCl was evaluated in MRS (bacilli) and M17 (cocci) media. Production of gas from glucose and arginine hydrolysis were also examined. For molecular identification, genomic DNA was extracted according to Pospiech & Neumann [ 12 ]. Strain differentiation was performed by repetitive sequence-based (rep-PCR) fingerprinting by using a (GTG)5 primer [ 13 ]. The master mix contained 4 µL of 5x buffer (InbioHighway, Argentina), 2 µL of 5mM dNTPs (Promega, Madison, WI, USA), 2 U of Taq polymerase (InbioHighway, Argentina), 1 µL of 50 ng DNA template, 2 µL of 10 mM primer (GTG)5 (Sigma-Merck, Germany), and 4 µL of 25 mM MgCl 2 . Polymerase chain reactions consisted of an initial denaturation at 94°C for 5 min, 30-cycle denaturation reaction at 94°C for 1 min, 1-min annealing at 40°C, 8-min extension at 65°C, and a final extension at 65°C for 10 min. DNA quantification was performed by using the NANO UV/Vis Spectrophotometer (MicroDigital Co., South Korea). Amplification reactions were carried out in a MyCycler thermal cycler (Bio-Rad, Richmond, CA, USA). PCR-products were separated by electrophoresis on a 1.5% (w/v) agarose at 40 V for 190 min and the molecular size marker used was 1 kb Plus DNA ladder (Waltham, Massachusetts, USA). Gels were stained with GelRed (Biotium Inc., CA, USA) and visualized with an ultraviolet light transilluminator (320 nm) using Image Scanner III (GE, Health Care, Pittsburgh, USA). Genomic DNA of selected isolates in each cluster was used for amplification of the almost full-length 16S rRNA gene fragment (~ 500pb) using the primers MLB and PLB [ 14 ] and sequenced at CERELA-CONICET by an ABI 3130 DNA sequencer (Applied Biosystems, CA, USA). rRNA gene sequence alignments were performed using the multiple sequence alignment method and identification queries were fulfilled by a BLASTn search in GenBank ( http://www.ncbi.nlm.nih.gov/GenBank/ accessed on 20 July 2023). 2.4. LAB Spoilage characteristics 2.4.1. Exopolysaccharides (EPS) production LAB ropiness was examined by the presence of a ropy condition after touching the colony with a loop as described by Ruiz Rodríguez et al. [ 15 ]. EPS from glucose and sucrose (4%) were investigated. EPS quantification was carried out by inoculation of 18 h subcultures (1%) into 50 mL modified MRS medium containing sucrose (100 g/L). After incubation (30°C for 72 h), the cultures were centrifugated (6000 g for 30 min) and the supernatants were collected. The extraction of EPS was performed according to the technique of van Geel-Schutten et al. [ 16 ] with minor modifications. Each supernatant was mixed with two-fold volume of cold ethanol (4°C). The supernatant-ethanol mixtures were kept for one night at 4°C and then centrifuged (2000 g for 15 min) to obtain EPS. The supernatants were removed, and EPS were subjected again to the same operation. Repeated rinsing of the precipitates was performed with absolute ethanol to eliminate any culture medium residues before drying at room temperature. The dried samples were diluted with ultrapure water and EPS levels were determined as mg glucose/L by phenol–sulfuric acid method [ 17 ]. Briefly, dried EPS was dissolved in deionized water until a final concentration of 5 mg/mL. To each 2 mL of EPS suspension, 50 µl of 80% (w/v) phenol and 5 mL of sulfuric acid (H 2 SO 4 ) was added quickly. The sample was incubated at room temperature for 45 min and the optical density at 490 nm was measured using a 96 U-well polystyrene microtitre (MtPs) plates (ExtraGene, Taiwan) and a microplate reader (SAFAS MP 96, Monaco) equipped with SAFAS software. The standard curve with linear regression of L-glucose (Sigma-Aldrich) at OD 490 was performed with Microsoft Excel 10. 2.4.2. Biogenic amine production Decarboxylase medium plates containing different precursor amino acids at a 1% final concentration (L-histidine monohydrochloride, tyrosine disodium salt, L-ornithine monohydrochloride and lysine monohydrochloride from Sigma-Aldrich, Germany) were used to test biogenic amines production according to Bover-Cid & Holzapfel [ 18 ]. 2.5. LAB Inhibitory activity 2.5.1. Antibacterial activity A semi-quantitative agar-spot-test was used to evaluate antibacterial activity of LAB isolates according to Fontana et al. [ 19 ]. In this study, native cell free supernatants (native-CFS) obtained by centrifugation (7000×g for 5min) were assayed against Gram-positive and Gram-negative bacteria used as target indicators (Table S1 ). Positive antimicrobial activity of LABs native-CFS were neutralized with 4N NaOH (Ciccarelli, Argentina) and treated with catalase (1000 U/ml) (Sigma-Aldrich, USA) to obtain treated-CFS. 2.5.2. Characterization of bacteriocin-like inhibitory substance (BLIS) against L. monocytogenes FBUNT To ascertain inhibitory activity, serial double dilutions of the inhibitory-treated cell-free supernatant (treated-CFS) were meticulously prepared using sterile distilled water. The agar-spot-test technique was subsequently employed for the determination of inhibitory potential [ 19 ]. After incubation, plates were examined for the appearance of clear zones, and inhibitory titers (AU/mL) were calculated according to the following formula: T = 1/V×D, where T is the inhibitory titer in AU/mL, V is the volume of the spotted supernatant (5 µl), and D is the maximum dilution factor with activity (halo presence). In addition, to confirm the proteinaceous nature of inhibitory substance/s each collected treated-CFS proteolytic enzymes were added including proteinase K, protease II, protease XIV, trypsin, pepsin and lysozyme (all from Sigma Aldrich, USA) at a final concentration of 0.1 mg/mL, incubated for 1 h at 37 ºC, heated at 90 ºC for 5 min to halt enzymatic reactions and the inhibitory activity was determined by the agar-spot-test method. Lat. curvatus CRL705 [ 20 ] and native-CFS were used as positive control. 2.5.3. Determination of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against L. monocytogenes FBUNT MIC was assessed in 96 U-well polystyrene microtitre (MtPs) plates (ExtraGene, Taiwan) as described by Ben Slama et al. [ 21 ]. LAB treated-CFS were subjected to serial dilutions in Brain Heart Infusion broth (BHI) (Britania, Argentina) to yield a final concentration ranging from 5 to 90% of 200 µL total volume in each well, then 10 µL of L. monocytogenes FBUNT at ∼10 9 CFU/mL were added to each well. MIC value was defined as the lowest concentration of treated-CFS without macroscopically visible growth. For the MBC determination, 10 µL of each well with no visible growth was poured on BHI agar plates and incubated at 30°C for 24 h. MBC was defined as the lowest concentration at which 99% of the bacteria were killed. 2.5.4. PCR detection of bacteriocin genes In this study, genomic DNA of positive control and LAB strains primers including specific and degenerate primers (Table S2 ) were designed and synthesized by Genbiotech (Buenos Aires, Argentina). PCR-amplifications were performed in 25 µl reaction mixture as described by Mechoud et al. [ 22 ]. PCR-products were separated by electrophoresis on 1% (w/v) agarose at 80 V for 45 min. DNA quantification and stained gels were visualized as previously stated. The resulting PCR of the strain Lat. sakei subsp. sakei 26 of presumed sak G product was purified with Wizard® SV Gel and PCR Clean-Up system (Promega Corporation, USA) according to the manufacturer’s instructions. DNA was sequenced at CERELA-CONICET through an ABI 3130 DNA sequencer using the forward or reverse primer. Analysis of DNA sequences was performed with the BLASTn search in GenBank program available at http://www.ncbi.nlm.nih.gov/GenBank/ accessed on 26 October 2023. 2.5.5. LAB antifungal activity The modified agar diffusion assay method described by Magnusson et al. [ 23 ] was used against autochthonous molds and yeast (Table S1 ). 2.6. LAB antibiotic resistance/susceptibility 2.6.1. Phenotypic antibiotic resistance and minimum inhibitory concentration (MIC) determination Minimum inhibitory concentrations (mg/mL) of the antibiotics ampicillin (AMP), clindamycin (CLI), chloramphenicol (CHL), erythromycin (ERY), gentamicin (GEN), kanamycin (KAN), tetracycline (TET), streptomycin (STR) and vancomycin (Van) were determined for LAB strains according to the ISO 10932:2010 standard. Epidemiological cut-off values based on the recommendation of the European Committee on Antimicrobial Susceptibility Testing (EUCAST) and EFSA-FEEDAP [ 24 ] were used. All antibiotics were purchased from Sigma-Aldrich (Missouri, USA) and ICN Biomedicals (California, USA). Each antibiotic was dissolved and filter-sterilized prior to its addition to mixed broth medium composed with 90% of ISM (Iso-Sensitest medium) from Oxoid (Thermofisher, Argentina) and 10% of MRS broth (Merck, Germany). The MIC was determined using the microdilution method described by the ISO/International Dairy Federation (IDF). The accuracy of susceptibility testing was monitored using quality control strain ( Lpb. plantarum ATCC14917) and two isolated strains ( E. faecium K15 and K19). 2.6.2. PCR detection of antimicrobial resistance genes . The presence of genes coding for antibiotic resistance (AR) in LAB strains phenotypically susceptible to antibiotics were evaluated through PCR reactions with the same reagents used above. The specific primers used, their target genes, the amplicon sizes, and the PCR protocol references used for gene detection are shown in Table S3 . The PCR amplifications were performed from total bacteria DNA obtained according to Pospiech & Neumann [ 12 ]. The PCR products were separated, gel staining and visualization were performed as explained above. 2.7. Statistical analysis All in-vitro assays were performed at least in duplicate with appropriate technical repetitions. Physicochemical and microbiological results were expressed as the mean value ± standard deviation of the data. Statistical analyses were performed with GraphPad Prism 7.04 Software using one-way ANOVA (nonparametric) followed by Tukey tests multiple comparison were P < 0.05 was considered as significatively different unless otherwise indicated. 3. Results 3.1. Physicochemical and microbial changes during the dry fermentation of Kaddid The evolution of physicochemical parameters and microbial population during the drying-fermentation of Kaddid prepared under laboratory conditions is shown in Table 1 . LAB growth reached maximal numbers between 3 and 7 days correlating with the lower pH values; the accumulation of acids (% of lactic acid) kept increasing, reaching ΔTTA of 1.8%±0.2 at the end of fermentation in agreement with the previously reported for traditional salted Kaddid [ 2 ]. In addition, it was observed that LAB and staphylococci are major populations in Kaddid , while coliforms were present in the meat (day 0) and then a remarkable reduction to undetectable level was observed. Table 1 Physicochemical and microbial (log CFU/g) changes during Kaddid dry-fermentation Sampling (days) 0 3 7 15 pH 5.58 ± 0.03 a 5.18 ± 0.02 b 5.28 ± 0.01 ab 5.45 ± 0.03 ab TTA (%) 0.40 ± 0.00 a 1.60 ± 0.10 ab 2.00 ± 0.10 b 2.20 ± 0.20 b Moisture (%) 67.41 ± 0.42 a 32.21 ± 3.12 b 25.71 ± 3.58 b 17.89 ± 1.83 b a w 0.87 ± 0.03 a 0.79 ± 0.03 a 0.71 ± 0.01 a 0.58 ± 0.02 d TAMB 4.45 ± 0.77 a 4.50 ± 0.54 a 4.34 ± 0.41 a 4.36 ± 0.25 a LAB 1.71 ± 0.18 a 2.53 ± 0.39 a 2.40 ± 0.33 a 2.22 ± 0.17 a Staphylococci 3.58 ± 0.37 a 3.35 ± 0.49 a 3.29 ± 0.27 a 2.71 ± 0.21 a Yeast & molds 2.43 ± 0.27 a 2.24 ± 0.18 a 2.10 ± 0.15 a < 1.00 b Total coliforms 2.73 ± 0.41 a < DL < DL < DL Salmonella spp. ND ND ND ND TTA: total titratable acidity; a W : water activity; TAMB: total aerobic mesophilic bacteria; LAB: lactic acid bacteria. Mean ± standard deviation; < DL: below the detection limit of the analytical method (10 CFU/g); ND: not detected (absence in 25 g). a−d : According to One-way ANOVA followed by Tukey test multiple comparison test ( P < 0.05) with 0.001 (99.9% confidence interval), means in the row sharing similar letters are not significatively different (ns: 0.1234), whereas group superscript different letters differ significantly. 3.2. Identification of LAB Bacteria isolates from the final stage of Kaddid drying-fermentation were investigated. Firstly, LAB isolates (29 bacilli/coccobacilli and 11 cocci) were subjected to a physiological preliminary identification (Table S4 ). The bacilli/coccobacilli (71.79%) cluster involved aerobic and facultative heterofermenter isolates that would be assigned to lactobacilli, Bacillus or Weissella genera with variable arginine hydrolysis, growth ability between 10 and 45°C in a wide range of pH except for pH 9.6 and resistant to high NaCl concentrations. Whereas homofermenter tetrads- and chain-forming cocci (10.25%) cluster growing between 10 to 45°C, up to 9.6% NaCl but not at pH > 8.0 were presumed as Pediococcus . However, the cluster of chain-forming homofermentative cocci (17.94%) developing at the same temperatures, in a wider pH range (4.0 to 9.6) and up to 6.5% NaCl were assigned to Enterococcus genus. Isolates were then sub-cultured (one single isolate from clustering profile of heterofermentative bacilli failed to grow and was discarded). Thus, a total of 39 strains showing growth were identified by (GTG)5-PCR and 16S rRNA sequencing yielding to a variable bands of molecular size ranging from 300 to 4000 bp corresponding to the genera Lactiplantibacillus , Latilactobacillus , Pediococcus , Enterococcus and Weissella (Fig. 1 ). Ascription of Kaddid isolates into species was based on the clusters derived from (GTG)5-PCR analysis; strains showing identical rep-PCR band patterns were considered as one rep-PCR biotype. Acordinly, isolates were grouped to 10 different (GTG)5-PCR biotypes. At least one representative from each biotype was identified by partial 16S rRNA gene sequencing. Biotype information from (GTG)5-PCR for LAB isolates (Fig. 1 ) allowed the association with Lactiplantibacillus (Lpb.) paraplantarum (K) 1, 2, 3, 4, 7, 8, 10, 12, 13, 14, 16, 17, 18, 20, 21, 31; Latilactobacillus (Lat.) sakei (K) 5, 25; Lat. sakei subsp. sakei (K) 22, 23, 24, 26, 27, 28, 29, 30; Pediococcus (P.) acidilactici (K) 34, 36, 37, 38; Enterococcus (E.) faecium (K) 6, 9, 11, 15, 19; E. hirae (K) 32, 33, Weissella (W.) cibaria (K) 35, 39 and Listeria (L.) monocytogenes FBUNT (40) used as indicator strain for antibacterial survey. 3.3. Screening of antimicrobial activity of LAB Inhibitory ability of LAB isolated from Kaddid was evaluated using Gram-positive/Gram-negative bacteria and yeast/molds as indicators (Table S1 ). High antagonistic activity against Gram positive indicator strains was found (Table 2 ). Inhibitory activity of LAB native-CFS was observed for Lpb. paraplantarum Lat. sakei /subsp. sakei and E. faecium against L. monocytogenes FBUNT, L. innocua CLIP74915 T , E. faecalis ATCC29212 and B. subtilis ATCC13453, whereas a lack of inhibitory activity was observed against Gram negative bacteria (data not shown). When LAB native-CFS were neutralized and treated with catalase (treated-CFS), several LAB strains maintained their inhibitory activity against Listeria ; titration of bacteriocin-like inhibitory substance (BLIS) showed a moderate antagonistic activity ranged between 200 and 400 AU/ml compared to the positive control Lat. curvatus CRL705 displaying an activity of 800 AU/ml. Moreover, antifungal activity against meat-borne molds showed most of LAB exhibiting a strong antagonism against Penicillium and Cryptococcus (Table 3 ). The highest antifungal activity was produced by Lpb. paraplantarum K1, K7, K13, K16, K17, K18 Lat. sakei K25, Lat. sakei subsp. sakei K23, K24, K28 as well as E. faecium K6, K11, K19 strains. However, assayed LAB failed to inhibit the growth of Aspergillus , Alternaria fungus and the commercial Saccharomyces cerevisiae strain. When antifungal compounds were investigated on LAB treated-CFS, a loss of LAB inhibitory activity was found, indicating that major antifungal compounds in CFS would be organic acids. Table 2 Frequency of antibacterial activity of LAB isolated from Kaddid against target indicators *BLIS: Bacteriocin–like inhibitory substance; **Positive controls of bacteriocin production. No inhibition detection (—); No determined (ND). Target strains were listed in table S1 . Identified LAB from Kaddid ( n =39) Antibacterial activity of LAB native-CFS against sensitive indicators BLIS* titration against L. monocytogenes FBUNT (AU/mL) L. monocytogenes FBUNT L. innocua L1PE L. innocua CLIP74915 T S. aureus ATCC29213 E. faecalis ATCC29212 B. subtilis ATCC13453 200 400 800 Lpb. paraplantarum (16 strains) 83.30% — 61.05% 5.55% 61.05% 49.95% K7, K8, K16 K18, K21 — Lat. sakei subsp. sakei (8 strains) 100% 12.50% 100% — 100% 100% K23, K24, K27, K28, K29, K30 K22, K26 — Lat. sakei (2 strains) 50% — 50% — 50% 50% K25 — P. acidilactici (4 strains) 75% — 25% — — — — E. faecium (5 strains) 100% — 60% — 60% 60% K6, K9 K11, K15, K19 — E. hirae (2 strains ) 100% — — — — — — W. cibaria (2 strains) 100% — 50% — — — — Lat. curvatus CRL705** 100% 100% ND ND ND ND 800 Lat. curvatus CRL1532** 100% 100% ND ND ND ND ND Table 3 Antifungal activity of LAB isolated from Kaddid against sensitive target strains CFS: cell free supernatant; — (no growth suppression), + (1–5 mm), ++ (5–10 mm), +++ (˃ 10 mm) of inhibition zone. Target strains were listed in table S1 . Identified LAB from Kaddid ( n =39) Inhibitory activity of LAB CFS against target indicators Penicillium S4J15 Penicillium C7J0 Rhizopus C5J0 Cryptococcus S3J0 Cryptococcus C3J7 Lbp. paraplantarum (16 strains) K1 (+++) K7 (++) K1, K7 (++) K13, K16, K17 K18 (+) K1, K7 (++) K16, K17, K18 (+) K1 (+++) K1, K7, K13, K16, K17, K18 (+++) Lat. sakei subsp. sakei (8 strains) — K28 (++) K23 (+++) K24 (++), K28 (+) K23, K24, K28 (+++) K23, K24, K28 (+++) Lat. sakei (2 strains) K25 (+) K25 (++) K25 (++) — K25 (+++) P. acidilactici (4 strains) — — — — — E. faecium (5 strains) K6, K11, K19 (+++) K6, K11 (+++) K19 (+) K6, K11, K19 (+) K6, K11, K19 (+++) K6, K11, K19 (+++) E. hirae (2 strains) — — — — — W. cibaria (2 strains) — — — — K35 (++) 3.3.1. Characterization of BLIS against L. monocytogenes FBUNT The different interference patterns on the inhibitory activity of tested BLIS from Kaddid lactobacilli are shown in Table 4 . When the effect of temperature and pH on lactobacilli BLIS was evaluated, a loss of inhibitory activity after treatments at 60 and 80°C was observed, while residual activity (200 AU/mL) after treatment at 100°C/30 min was found for Lat. sakei K25 and Lat. sakei subsp. sakei K26 and after 121°C/15 min for Lpb. paraplantarum K21. Even lower, all tested strains preserved their antibacterial activity after storage at 5°C for 7 d. In addition, highest activity (400 AU/mL) was shown at pH 6.5 that was retained up to pH 4.5 (200 AU/mL), but a lack of inhibition was found at pHs 2, 8 and 10. Results indicated that BLIS from Lpb. paraplantarum K18 and K21, Lat. sakei K25 and Lat. sakei subsp. sakei K26 strains were inactivated by protease XIV and proteinase K; protease V was effective at inactivating BLIS from Lat. sakei K25 and Lat. sakei subsp. sakei K26, whereas lysosyme, pepsine and trypsine were unable to inactivate BLIS activity of any of the assayed lactobacilli (Table 4 ). Additionally, microdilution method used to determine the MIC of BLIS against L. monocytogenes FBUNT, showed 20% (80 AU/mL) of BLIS as effective for all tested strains with variable MBC (between 70 and 80%); a lower inhibitory potency of Kaddid strains (4xMIC) compared to Lat. curvatus CRL705 (MBC>100%) was detected. Furthermore, the presence of bacteriocin encoding genes was also evaluated (Table 4 ; Fig. S1 ). The results revealed that strains Lpb. paraplantarum K18 and Lat. sakei subsp. sakei K26 exhibited comparable genetic profiles for bacteriocin-related genes; although both strains demonstrated the presence of amplified products for skg A (sakacin G) and sppA (sakacin P); Lat. sakei K25 and Lat. sakei subsp. sakei K22 showed positive results only for skg A and spp A respectively. Structural genes for curvacin A ( sap A) and sakacin Q ( spp Q) were not present in the tested lactobacilli from Kaddid and Lpb. paraplantarum K21 lack the encoding genes for sakacin. Table 4 BLIS activity characterization and presence of bacteriocin genes in selected lactobacilli from Kaddid Lpb. paraplantarum Lat. sakei Lat. sakei subsp. sakei Lat. curvatus CRL705 (control) K18 K21 K25 K22 K26 Control MRS, pH 6.5/30°C 400 400 400 400 400 800 Temperature* 45°C/60 min 400 400 400 400 400 600 60°C/60 min — — — — — 600 80°C/30 min — — — — — 400 100°C/30 min — — 200 — 200 200 121°C/15 min 200 200 — 200 — 200 5°C/ 7 d 200 200 200 200 200 200 pH* 4.5/30 ºC 200 200 200 200 200 400 2, 8, 10/30 ºC — — — — — — Enzymes ** Lysozyme/pepsine/trypsine + + + + + + protease V + — — + — + protease XIV — — — + — — proteinase K — — — + — — MIC (%) 20 (80 AU/mL) 20 (80 AU/mL) 20 (80 AU/mL) 20 (80 AU/mL) 20 (80 AU/mL) 5 (40 AU/mL) MBC (%) 80 70 70 70 80 ˃100 Bacteriocin gene(s ) skg A , spp A ND skg A , spp A skg A , spp A spp A , spp Q *Results are indicated as AU/mL; ** (+): presence of inhibition halo (≥ 6 mm); (-) absence of inhibition halo. Lat. sakei CRL1862 and Lat. curvatus CRL1537 used as control for curvacin A ( sap A) and sakacin P ( spp A) genes (Fontana et al., 2015). and sakacin G ( skg A), this study. 3.4. Spoilage potential of Kaddid lactobacilli The phenotypical detection of gas, exopolysacharides (EPS) and biogenic amines (BA) production by Kaddid selected strains were investigated (Table 5 ). All the tested strains displayed gas production from gluconate, while only Lat. sakei subsp. sakei K22 produced minimal amount of gas from glucose in correlation with their facultative heterofermentative character. Even in lower abundance compared to positive control, a ropy phenotype from glucose for K25, K22 and K26 lactobacilli was detected while only K22 strain presented ropiness from sucrose. The ropiness phenotype for Lpb. paraplantarum K18 and K21 strains was barely detectable while that of Lat. sakei K25 and Lat. sakei subsp. sakei K22 and K26 were more detectable (Table 5 ). Even EPS production values were lower than that from W. cibaria BK19 (2464.62 ± 113.34 mg/L) used as control, EPS production by K22 from sucrose exhibited the highest value of 574.62 ± 2mg/L. No significant differences ( P < 0.05) were found between the remaining strains with lower EPS production values. In addition, when Kaddid selected lactobacilli were tested for their decarboxylase ability, none of the assayed strains were observed to produce bioactive amines (data not shown). Table 5 Spoilage potential of selected lactobacilli from Kaddid Lactobacilli Gas from Ropiness from EPS (as mg of glucose/L) * glucose gluconate glucose sucrose Lpb. paraplantarum K18 - + - - 138.37 ± 05.36 a Lpb. paraplantarum K21 - + - - 327.12 ± 22.54 a Lat. sakei K25 - + + - 258.37 ± 11.48 a Lat. sakei subsp . sakei K22 + + ++ + 574.62 ± 24.97 d Lat. sakei subsp . sakei K26 - + + - 452.12 ± 23.88 a Lpb. plantarum CRL972 - + ND ND ND Lat. sakei CRL1407 ND ND +++ +++ ND W. cibaria BK19 ND ND +++ +++ 2464.62 ± 113.34 f Positive test (+, ++, +++); negative test (-); not determined (ND). Lpb. plantarum CRL972 and Lat. sakei CRL1407 (CERELA culture collection) and W. cibaria BK19 were used as positive control for gas and EPS production, respectively. *: EPS from 100g/L of sucrose; a−f : According to One-way ANOVA followed by Tukey test multiple comparison test ( P < 0.05) with 0.001 (99.9% confidence interval), means in the same column sharing similar letters are not significatively different (ns: 0.1234), whereas group superscript different letters differ significantly. 3.5. Antibiotic resistance and detection of AR gene(s) The analysis was performed following the cut-off value and the indications given by EFSA [ 24 ]. The MICs determination according to ISO/IDF of eight antibiotics was examined in the selected Kaddid lactobacilli and two isolates of E. faecium (K15 and K19), (Table 6 ). Kaddid lactobacilli revealed an unexpected high resistance to TET, a double resistance TET/STR for Lat. sakei K25, whereas Lpb. paraplantarum K18 was the only strain with no phenotypical antibiotic resistance and all lactobacilli were susceptible to the rest of the evaluated antibiotics. Lat. sakei K25 displayed MIC values for STR three-fold the cut-off value (64 µL/g), whereas a MIC of four-fold the cut-off value (8 µL/g) was found for TET. When the presence of AR gene(s) was investigated by PCR, tet M gene was found in all strains even in Lpb. paraplantarum K18 that was phenotypically sensitive to TET (Table 6 ; Fig. S2 ). Table 6 MICs distribution and antibiotic resistance (AR) genes among Kaddid selected lactobacilli AMP CLI CHL ERY GEN KAN STR TET VAN AR genes Lpb. paraplantarum Cut-off value 2 4 8 1 16 64 n.r. 32 n.r K18 MIC <0.032 <0.032 0.125 <0.016 128 tet M K21 0.125 <0.032 0.125 128 tet M Lat. sakei K25 Cut-off value 2 4 8 1 16 64 64 32 n.r MIC 0.5 <0.032 0.125 128 tet M Lat. sakei subsp . sakei K22 MIC 0.125 <0.032 0.125 <0.016 0.5 128 tet M K26 0.25 <0.032 0.125 <0.016 2 128 tet M E. faecium Cut-off value 2 4 16 4 32 1024 128 4 4 K15 MIC 4 1 0.125 8 32 16 128 <0.125 1 K19 4 16 0.125 8 32 1024 256 64 < 0,25 Cut-off values were proposed by the EFSA-FEEDAP (2018) and MIC are expressed in µg/mL; numbers in bold indicate antibiotic resistance; n.r : not required. 4. Discussion During Kaddid production a lactic fermentation takes place as reported in other dry-cured and salted meat products such as “ Lacón ” [ 25 ]. The small size and thickness of Kaddid strips represent a large surface-to-weight ratio favoring dehydration during post-salting and drying stages leading to great moisture loss and a w decrease. The presence of high salt concentration helps to draw water and sugars out of meat tissue; salt uptake and water exudation from meat are mutually dependent [ 26 ]. On the other hand, microbiological traits at 15 days showed LAB and staphylococci as major populations in Kaddid . LAB presence was found to be higher than that reported for Tunisian Kaddid [ 27 ], this correlates with the controlled conditions of laboratory-made Kaddid . The presence of coagulase-negative staphylococci agrees with the abundant presence of Staphylococcus genus in salted/cured meat [ 28 ], which are tolerant to high salt concentration. In accordance, several LAB species were reported to adapt to high NaCl concentration, particularly Lat. sakei that was identified from salted anchovies resisting up to 15% of NaCl [ 29 ]. The reduction of coliforms level was in correlation with the increased LAB growth and acid production, representing a good hygienic indicator after preparation/handling of meat for Kaddid production. Results of LAB molecular identification agrees with those from high salt-containing meat products [ 1 , 11 ]. In fact, the presence of the phylum Firmicutes, including Lactobacillaceae , Leuconostocaceae and Enterococcaceae families was widely reported from dry-salted meat and sea food products [ 29 , 30 , 31 ]. The dominance of lactobacilli species at the final stage of the dry-ripening salted and/or cured dry-meat products is in correlation with their ability to adapt to high NaCl during fermentation, processing and storage. Indeed, Lat. sakei identification was reported for Algerian El-Guedid [ 1 ]. Furthermore, Lat. sakei species was widely identified as predominant from many dry-fermented sausages across the Mediterranean region [ 8 , 32 , 33 , 34 ]. The presence of Lat. sakei and Lat. sakei subsp. sakei in laboratory-made Kaddid could be assigned to the dry-ripening temperature (22 ± 1°C) used, in agreement with the detection of Lat. sakei and Lat. curvatus in sausages ripened at low temperatures while Lpb. plantarum was dominant at higher temperatures (> 25°C) as was reported by Lücke et al. [ 35 ]. The presence of Lpb. paraplantarum was also reported from Andalusian traditional spontaneous fermented sausages [ 36 ], whereas Weissella was isolated from traditional homemade Kaddid prepared under Saharan conditions in South-Western Algeria [ 11 ]. In addition, added spices to raw meat may harbor a diverse and abundant microbial community with LAB species such as Weissella, Pediococcus and Enterococci [ 37 ]. It is known that metabolic compounds produced by LAB, including organic acids, hydrogen peroxide and bacteriocins, can exert antimicrobial effect against a range of pathogens and contaminants [ 38 ]. The highest inhibitory activity (400 AU/mL) after CFS titration against L. monocytogenes FBUNT was observed for Lpb. paraplantarum K18 and K21, Lat. sakei K25, Lat. sakei subsp. sakei K22, K26 and E. faecium K11, K15, K19 strains and may be attributed to bacteriocin production. When antifungal activity against meat-borne molds were analyzed, Lat. sakei and Lpb. paraplantarum strains showed high activity against Penicillium in agreement with that of Lat. sakei subsp. ALI033 isolated from Kimchi which was suggested to be most likely due to organic acids production [ 39 ]. Similarly, Lat. sakei isolated from Sucuk , a Turkish dry-fermented sausage was reported to exert antifungal activity against Penicillium and Aspergillus fungi [ 40 ]. Lpb. paraplantarum antifungal activity found here is compatible to that reported for a non-starter dairy strain [ 41 ]. Based on the antimicrobial traits of Kaddid isolates, Lpb. paraplantarum K18 and K21, Lat. sakei K25 and Lat. sakei subsp. sakei K22, K26 strains were selected for their promising features mostly due to their ability to inhibit L. monocytogenes FBUNT. In contrast, E. faecium strains were not included because of their detrimental traits, virulence factors and antibiotic resistance were not included [ 42 , 43 ]. Since bacteriocins from LAB are small peptides (<5 kDa), they are usually thermostable [ 44 , 45 ]. However, although it was strain dependent, we found a general trend showing a decrease in bacteriocin activity with increasing temperature. BLIS produced by lactobacilli strains from Kaddid , resisted high temperature and pressure during sterilization, indicating they were heat-stable peptides. Similar results were reported for Lat. sakei and Lat. curvatus from fermented meat products and Lpb. plantarum from dairy products, which remained active after 100°C/2 h and 80°C/30 min respectively [ 46 , 47 , 48 ]. Moreover, BLIS inhibitory activity showed to be higher after exposure to pH 6.5 (400 AU/mL) compared to pH 4.5 (200 AU/mL), Similarly, Lpb. plantarum and Lat. sakei subsp. sakei 2a of dairy and meat origin, showed maximal bacteriocin production at somewhat acidic pH [ 48 , 49 ]. Confirmation of proteinaceous nature of bacteriocin is essential for the characterization of new antimicrobial peptides. Indeed, inactivation of BLIS inhibitory activity by proteolytic enzymes clearly showed that the antimicrobial substance was of proteinaceous nature; even though the presence of the inhibition halo after enzyme treatment could also suggest other products such as acids or glycoactive compounds as responsible inhibitors. Lat. curvatus CRL705 used as control showed higher inhibitory activity (AU/mL) in agreement with the five protein-encoding genes (lactocin 705, sakacin P, sakacin Q, sakacin X, and sakacin T) responsible for bacteriocin production reported by genome sequencing [ 20 ]. Although the results obtained from MIC and MBC of BLIS against L. monocytogenes FBUNT are consistent with previous bacteriocin reported from Lactobacillus [ 50 ]; antimicrobial compound MICs against the same pathogen species can be variable depending on the strain, the antimicrobial preparation and experimental design used [ 51 ]. In addition, Class IIa anti-listeria bacteriocins potency would be influenced by the cell-envelope lipid composition of target membranes or differences in the three-dimensional structures of antimicrobial peptides which would determine the varied values of reported MICs [ 45 ]. The presence of the spp A gene for sakacin P but not sap A for curvacin A detected in Lat. sakei subsp. sakei and Lpb. paraplantarum strains agrees with that described for LAB species isolated from different Argentina meat products [ 19 ]. Sakacin G is known to possess duplicate structural genes skg A1 and skg A2 [ 52 ]. The results of sakacin G assessment revealed a PCR fragment of approximately 200 bp, which doubled the anticipated size with the employed primers in this study [ 22 ]. To validate the identity of this gene, the nucleotide sequence of the PCR fragment amplified from the exemplary producer Lat. sakei subsp. sakei K26, was sequenced. The analysis of the nucleotide sequence showed 100% similarity to the pre-bacteriocin skg A2 gene (accession number FJ621568). Results from this study correlated with the sakacin P, sakacin G encoding genes reported for Lat. sakei CWBI-B1365 from raw poultry meat, Lat. curvatus ACU-1 from fermented sausages and Lat. sakei R1333 from smoked salmon with high anti- Listeria activity [ 22 , 52 , 53 ]. These bacteriocins belong to class IIa which show strong anti-listeria inhibitory effect as well as other food spoilage and pathogenic bacteria; these small peptides have received much attention due to their generally recognized as safe (GRAS) status, their high biological activity, and their heat stability [ 45 ]. The presence of bacteriocin-related genes does not necessarily indicate the expression of these peptides; horizontal gene transfer mechanisms can explain the variety of genes and the production of multiple bacteriocins by the same culture [ 54 ]. On the other hand, in meat products the absence of gas, exopolysaccharides (EPS) and biogenic amines (BA) production by starter cultures must be avoided, since they would lead to an indication of spoilage. Although gas production from gluconate was observed for all tested Kaddid lactobacilli, it did not represent a spoilage risk (it was not included among Kaddid preparation additives), whereas gas from glucose was produced by Lat. sakei subsp. sakei K22. The production of ropiness was more notable using glucose than sucrose; Lpb. paraplantarum K18 and K21 were unable to exhibit a ropy phenotype in correlation with their lowest EPS production level. EPS production by LAB showed average yield ranging between 25 to 9800 mg/L [ 55 , 56 ] using different sugar as substrates. Results from this study are in coincidence with the EPS-forming Lat. sakei KS-82 from Turkish Sucuk and the high molecular weight EPS produced by the probiotic Lpb. paraplantarum BGCG11 strains [ 57 , 40 ]. Although EPS-producing LAB are of technological interest due to their ability to modify food texture and delay color oxidation (Rodríguez-Sánchez et al., 2021), they are considered undesirable for meat products due to slime production. In addition, no BA production was detected among tested lactobacilli, in correlation to that reported for Lat. sakei strains from Italian sausages [ 58 ]. Safety concerns on the use of LAB strains carrying antibiotic resistance (AR) genes have been raised, as they can transfer them to pathogenic bacteria through horizontal gene transfer mechanisms. Their presence on LAB to be used as autochthonous starter culture might constitute a possible public health hazard, since fermented foods are important vehicles for enormous amounts of living bacteria to enter human body [ 59 ]. Results revealed high resistance to TET among Kaddid selected lactobacilli. Indeed, TET is among the most widely reported antibiotics for resistant lactobacilli from fermented meats. Lat. sakei and Lpb. plantarum strains from fermented sausages of different origins showed a high incidence of TET resistance and in a less abundance to ERY [ 8 , 33 ]. However, high resistance to STR was also described for Lactobacillaceae from Mediterranean fermented sausages involving Lat. sakei as dominant species [ 8 ]. TET resistance of Lpb. paraplantarum K21 disagrees to that reported for a dairy strain [ 60 ]. The prevalence of Kaddid lactobacilli resistant to TET and STR was supported by the common use of a penicillin/streptomycin complex and/or oxytetracycline for ovine livestock farming in Algeria. These antimicrobials are systematically prescribed to prevent uterine and respiratory infections in postpartum ewes, as well as respiratory and gastrointestinal infections in lambs [ 61 ]. Widely distributed resistance to aminoglycosides (STR, KAN, GEN) among lactobacilli is likely to be intrinsic as reported by Anisimova & Yarullina [ 62 ]. This naturally occurring AR is considered relatively safe, since no horizontal transfer between isolates or species was reported [ 63 , 64 ]. Similarly, an intrinsic resistance of lactobacilli to vancomycin was also reported [ 65 ]. The high sensitivity to ERY, CLI and CHL observed in lactobacilli from Kaddid agrees with the general susceptibility of lactobacilli to antibiotics inhibiting protein synthesis and resistance to aminoglycosides [ 66 ]. In contrast, Lat. sakei strains from Mediterranean sausages were phenotypically resistant to AMP, GEN, KAN, CLI and CHL but not to ERY [ 8 , 33 ]. On the other hand, the remarkable incidences of AR in enterococci found in this study is in agreement with that reported in animal’s feces and raw and fermented meat strains [ 42 , 67 , 68 ]. The importance of food enterococci as a reservoir of antibiotic resistance genes and the potential for their genetic transfer to human strains following consumption of uncooked or undercooked contaminated meat is of high concern. When the presence of AR genes was assessed, tet M gene was predominantly found in Kaddid strains. As reported by Fontana et al. [ 8 ], this gene was associated with different MIC values (32–256 µg/mL) of lactobacilli strains. Indeed, increasing tet M transcript levels were correlated with increased MIC values, suggesting that the expression of this gene is dependent on TET concentration [ 69 ]. In this study, a MIC value of 64 µg/mL was found for K21, K22, K25 and K26 lactobacilli strains expressing tet M gene, however Lpb. paraplanatarum K18 had a low MIC (4 µg/mL) and a sensitive phenotype to TET also expressing tet M gene. The correlation between MIC and genetic determinants incidence is strictly strain dependent, however the association between high MICs and the existence of at least one genetic determinant for TET resistant strains was also described [ 8 ]. The abundance of TET resistance genes in pig feces microbiome and, consequently in the raw meat used for fermented sausages was also reported [ 70 , 71 ]. Furthermore, even when Lat. sakei K25 was categorized as phenotypically resistant, none of the Kaddid isolates showed the presence of genes associated with STR resistance suggesting that these genes were not involved in this isolate’s resistance. Similarly, Lpb. paraplantarum K18 was phenotypically susceptible to TET and showed to harbor tet M gene. These discrepancies between the resistance phenotype and genotype may be the result of defective expression of resistance genes [ 62 ]. Altogether, the results from this study confirm the prevalence of tet M gene representing the most widespread resistance determinant in Kaddid strains. Furthermore, it was reported that tet M is located in plasmids suggesting that it has the potential to move between different organisms with an encoding ribosomal protection protein catalyzing the release of tetracycline from the ribosome [73]. Therefore, TET resistance in bacterial strains for food and agricultural applications always constitutes a risk of AR resistance genes spread in the environment. 5. Conclusions The results of this study suggest promising use of autochthonous LAB strains for production of traditional Kaddid . Among evaluated lactobacilli, The strains Lpb. paraplantarm K18 and K21 exhibited desirable in vitro functional properties namely the inhibitory potential against L. monocytogenes , low risk as spoilers and free of antibiotic resistance, therefore considered as good candidates to be used as a successful autochthonous functional starter culture to improve Kaddid quality and safety. Further investigation to assess their technological and functional performance is currently undertaken. Declarations Supplementary Materials: Tables S1, S2, S3 and S4; Figure S1 and S2. Acknowledgments: Kamel’s Boubakri PhD scholarship long stay at CERELA (PNE 2018/2019, N° 256) was supported by the Algerian Ministry of high education and scientific research and the university of Médéa, Algeria. Theauthors thank Dr. Nadia E. Suarez from the sequencing service at CERELA for their help in biomolecular interpretations and Mouna El Hassani PhD at the university of Medea for reviewing the English language of the manuscript. Author contributions Kamel Boubakri: Conceptualization, Formal analysis, Data curation, Investigation, Methodology, Software, Validation, Visualization, Writing original draft. Tayeb Idoui: Conceptualization, Project administration, Validation. Writing review & editing. Cecilia Aristimuño Ficoseco: Formal analysis, Methodology, Resources, Software, Supervision, Validation. Franco J. Segli: Methodology, Resources. Patricia Castellano: Funding acquisition, Methodology, Resources. Lucila Saavedra : Funding acquisition, Methodology, Resources. Writing review & editing . Graciela M. Vignolo: Conceptualization, Data curation, Funding acquisition, Investigation, Methodology, Project administration, Resources, Supervision, Validation. Writing review & editing. All authors have read and agreed to the published version of the manuscript. Data Availability: The datasets generated during the current study are available in the article and in the online supplementary material. Funding: No funding was received for conducting this study. Conflict of Interest: The authors declare no competing interests. Financial interests: The authors have no relevant financial or non-financial interests to disclose. Ethical Approval Statement: The paper reflects the authors’ own research and analysis in a truthful and complete manner. Patient consent statement: not applicable Permission to reproduce material from other sources: not applicable References Bader R, Becila S, Ruiz P, Djeghim F, Sanah I, Boudjellal A, Gatellier P, Portanguen S, Talon R, Leroy S. Physicochemical and microbiological characteristics of El-Guedid from meat of different animal species. 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Determination of antibiotic resistance of lactic acid bacteria isolated from traditional Turkish fermented dairy products. Iranian J Vet Res. 2018; 19: 53–56. https://doi.org/10.22099/ijvr.2018.4769 Sharma K, Sharma N, Sharma R. An evaluation of in-vitro potential of novel Lactobacillus Paraplantarum KM0 (KX671558) strain isolated from milk. Proc Indian Nat Sci Acad. 2017; 83(3): 689–699. https://doi.org/10.16943/ptinsa/2017/49063 Azzi O, Lai F, Tennah S, Menoueri MN, Achek R, Azara E, Tola S. Spa-typing and antimicrobial susceptibility of Staphylococcus aureus isolated from clinical sheep mastitis in Medea province, Algeria. Small Rum Res. 2020; 192: 106168. https://doi.org/10.1016/j.smallrumres.2020.106168 Anisimova EA, Yarullina DR. (2019) Antibiotic resistance of Lactobacillus strains. Current Microbiol. 2019; 76(5): 1–10. https://doi.org/10.1007/s00284-019-01769-7 Shao Y, Zhang W, Guo H, Pan L, Zhang Z, Sun T. Comparative studies on antibiotic resistance in Lactobacillus casei and Lactobacillus plantarum . Food Control. 2015; 50: 250–258. https://doi.org/10.1016/j.foodcont.2014.09.003 Wu M, Tarrah A, Ghion G, Pakroo S, Giacomini A, Corich V. A critical issue on microbiological cut-off value of ampicillin resistance in Lactiplantibacillus plantarum . J Appl Microbiol. 2023; 134(3): lxad050. https://doi.org/10.1093/jambio/lxad050 . Abriouel H, Casado Muñoz MC, Lerma LL, Montoro BP, Bockelmann W, Pichner R, Kabisch J, Cho G-S, Franz CMAP, Gálvez A, Benomar N. New insights in antibiotic resistance of Lactobacillus species from fermented foods. Food Res Int. 2015; 78: 465–481. https://doi.org/10.1016/j.foodres.2015.09.016 Ammor MS, Flórez AB, Mayo B. Antibiotic resistance in non-enterococcal lactic acid bacteria and bifidobacteria. Food Microbiol. 2007; 24: 559–570. https://doi.org/10.1016/j.fm.2006.11.001 Różańska H, Lewtak-Piłat A, Osek J. Antimicrobial resistance of Enterococcus faecalis isolated from meat. Bull Vet Inst Pulawy. 2015; 59: 229–233 https://doi.org/10.1515/bvip-2015-0034 Aristimuño Ficoseco C, Mansilla FI, Maldonado NC, Miranda H, Nader-Macias MEF, Vignolo GM. Safety and growth optimization of Lactic Acid Bacteria isolated from feedlot cattle for probiotic formula design. Front Microbiol. 2018; 9: 2220. https://doi.org/10.3389/fmicb.2018.02220 Comunian R, Daga E, Dupré I, Paba A, Devirgiliis C, Piccioni V, Perozzi G, Zonenschain D, Rebecchi A, Morelli L, De Lorentiis A, Giraffa G. (2010) Susceptibility to tetracycline and erythromycin of Lactobacillus paracasei strains isolated from traditional Italian fermented foods. Int J Food Microbiol. 2010; 138: 151–156. https://doi.org/10.1016/j.ijfoodmicro.2009.11.018 Monger XC, Gilbert AA, Saucier L, Vincent AT. Antibiotic resistance: From pig to meat. Antibiotics. 2021; 10(10): 1209. https://doi.org/10.3390/antibiotics10101209 Uezen JD, Aristimuño Ficoseco C, Nader-Macías MEF, Vignolo GM. Identification and characterization of potential probiotic lactic acid bacteria isolated from pig feces at various production stages. Canadian J Vet Res. 2023; 87(2): 127–145. PMCID:PMC10069149 Gevers D, Masco L, Baert L, Huys G, Debevere J, Swings J. Prevalence and diversity of tetracycline resistant Lactic Acid Bacteria and their tet genes along the process line of fermented dry sausages. Syst Appl Microbiol. 2003; 26(2): 277–283. https://doi.org/org/10.1078/072320203322346137 Additional Declarations No competing interests reported. Supplementary Files FigS1.bacteriocinsgels.docx FigS2.antibioticgenes.docx TableS1.Targetstrains.docx TableS4.PhysiologicalandBiochemicalclustering.docx Cite Share Download PDF Status: Published Journal Publication published 23 Apr, 2025 Read the published version in Discover Food → Version 1 posted Editorial decision: Revision requested 13 Nov, 2024 Reviews received at journal 13 Nov, 2024 Reviewers agreed at journal 06 Nov, 2024 Reviewers agreed at journal 05 Nov, 2024 Reviewers agreed at journal 05 Nov, 2024 Reviews received at journal 20 Oct, 2024 Reviewers agreed at journal 15 Oct, 2024 Reviewers agreed at journal 01 Oct, 2024 Reviewers invited by journal 24 Sep, 2024 Editor assigned by journal 09 Sep, 2024 Submission checks completed at journal 05 Sep, 2024 First submitted to journal 30 Aug, 2024 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-5005817","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":362637150,"identity":"9de07b68-c9f2-470c-a328-602a27ff7aa9","order_by":0,"name":"Kamel Boubakri","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA9klEQVRIiWNgGAWjYBACCQkGBmYGhgQGNvbmww8+AEXY2InWwnMszXAGSAszsVoYJHIMpHlAQoS0SM5uPiZdUJOWz8eQYGBs82ubPB8zA+OHjzm4tUjLHEuTnnEsx7KN4UDC49y+24ZtzAzMkjO34dYiJ5FjJs3DVmHAxthwwDi35zYjUAsbMy9BLf+AWpgZG6Qte27bE9QiDdLC25ZjwAYMKWmGH7cTCWqRnHMs2XpmX5oBGw8bm2Fvw+3kNmbGZrx+kbjdfPB2wbdkA/n57z8/+PHntu389uaDHz7i0YIKGNvAZAOx6kHgDymKR8EoGAWjYKQAANcdSRPcfemFAAAAAElFTkSuQmCC","orcid":"","institution":"Université Yahia Farès de Médéa","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Kamel","middleName":"","lastName":"Boubakri","suffix":""},{"id":362637151,"identity":"456f7b48-912b-4f20-883f-2976ff2ec4a2","order_by":1,"name":"Tayeb Idoui","email":"","orcid":"","institution":"Université de Jijel","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Tayeb","middleName":"","lastName":"Idoui","suffix":""},{"id":362637152,"identity":"5d6ad964-a1a6-4b58-ac39-edb2044b142e","order_by":2,"name":"Cecilia Aristimuño Ficoseco","email":"","orcid":"","institution":"CONICET","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Cecilia","middleName":"Aristimuño","lastName":"Ficoseco","suffix":""},{"id":362637153,"identity":"391d7cc2-4fee-420b-86ac-402595312ffa","order_by":3,"name":"Franco J. Segli","email":"","orcid":"","institution":"CONICET","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Franco","middleName":"J.","lastName":"Segli","suffix":""},{"id":362637154,"identity":"a40b077e-2a5d-4ac1-942d-598bc08d6234","order_by":4,"name":"Patricia Castellano","email":"","orcid":"","institution":"CONICET","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Patricia","middleName":"","lastName":"Castellano","suffix":""},{"id":362637155,"identity":"b15ad6bd-1b4c-40be-b0d3-9b686feddfdc","order_by":5,"name":"Lucila Saavedra","email":"","orcid":"","institution":"CONICET","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Lucila","middleName":"","lastName":"Saavedra","suffix":""},{"id":362637156,"identity":"444931a2-ff8e-4abf-a23f-7858c2bce17f","order_by":6,"name":"Graciela M. Vignolo","email":"","orcid":"","institution":"CONICET","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Graciela","middleName":"M.","lastName":"Vignolo","suffix":""}],"badges":[],"createdAt":"2024-08-30 19:39:55","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-5005817/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5005817/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s44187-025-00347-0","type":"published","date":"2025-04-24T00:00:00+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":66101455,"identity":"53a51d15-68f5-458a-958b-a46adf813e1f","added_by":"auto","created_at":"2024-10-07 17:09:29","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":127588,"visible":true,"origin":"","legend":"\u003cp\u003eDendrogram of cluster analysis of rep-PCR fingerprints of LAB isolated from Kaddid. (GTG)5-RAPD profiles including the following biotypes: 18-Lactiplantibacillus (Lpb.) paraplantarum 99.79% (accession: CP053337.1); 25-Latilactobacillus (Lat.) sakei (99.41%) (accession: JQ653151.1); 34-Pediococcus (P.) acidilactici 96.19% (accession: CP053421.1); 24-Lat. sakei subsp. sakei 81.64% (accession: NR_042443.1); 11-Enterococcus (E.) faecium 99.79% (accession: AP019394.1) ; 15-E. faecium \u0026nbsp;97.47% (accession: AP019394.1) ; 13-Lpb. paraplantarum 98.44% (accession: CP053337.1) ; 33-E. hirae \u0026nbsp;99.36% (accession: CP055232.1) ; 35-Weissella (W.) cibaria \u0026nbsp;99.19% (accession: MN700179.1); 39-W. cibaria 99.75% (accession: MT613505.1) \u0026nbsp;; 32- E. hirae 99.79% (accession: CP055232.1); 40- Listeria (L.) \u0026nbsp;monocytogenes FBUNT 99.23% (accession: CP054846.1); MW: Molecular weight marker (1 kb DNA ladder). Merged RAPD-PCR patterns were grouped by means of the Pearson's mean correlation coefficient and UPGMA cluster analysis with GelJ program.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-5005817/v1/22a31d1c92c2304ae2653592.png"},{"id":90039418,"identity":"66f58df2-b4d2-4144-bd3a-84fba7abda40","added_by":"auto","created_at":"2025-08-27 16:34:14","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2460538,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5005817/v1/342c7cda-49c9-4fe1-96fd-d84053198b3c.pdf"},{"id":66101458,"identity":"3ffca58c-71c5-4b77-93d0-ce4df9541a6b","added_by":"auto","created_at":"2024-10-07 17:09:29","extension":"docx","order_by":6,"title":"","display":"","copyAsset":false,"role":"supplement","size":604835,"visible":true,"origin":"","legend":"","description":"","filename":"FigS1.bacteriocinsgels.docx","url":"https://assets-eu.researchsquare.com/files/rs-5005817/v1/34ccf3e05b8137582672f9ab.docx"},{"id":66101691,"identity":"3e3fa43f-cd2e-4cfa-bf84-bcf762560a50","added_by":"auto","created_at":"2024-10-07 17:17:29","extension":"docx","order_by":7,"title":"","display":"","copyAsset":false,"role":"supplement","size":221787,"visible":true,"origin":"","legend":"","description":"","filename":"FigS2.antibioticgenes.docx","url":"https://assets-eu.researchsquare.com/files/rs-5005817/v1/66eed279b672fe9c1f97ba3c.docx"},{"id":66101456,"identity":"a45cac43-cab9-474c-adb2-3febee418ea1","added_by":"auto","created_at":"2024-10-07 17:09:29","extension":"docx","order_by":8,"title":"","display":"","copyAsset":false,"role":"supplement","size":25205,"visible":true,"origin":"","legend":"","description":"","filename":"TableS1.Targetstrains.docx","url":"https://assets-eu.researchsquare.com/files/rs-5005817/v1/922fb037dae61204740aa81d.docx"},{"id":66101457,"identity":"656e5d33-0ca9-4dcd-b86f-03a1438c84d0","added_by":"auto","created_at":"2024-10-07 17:09:29","extension":"docx","order_by":11,"title":"","display":"","copyAsset":false,"role":"supplement","size":17528,"visible":true,"origin":"","legend":"","description":"","filename":"TableS4.PhysiologicalandBiochemicalclustering.docx","url":"https://assets-eu.researchsquare.com/files/rs-5005817/v1/cababebd2fab6b40a5892477.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Autochthonous Lactic Acid Bacteria from Kadidd as Functional Starter Culture to Improve quality and safety","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003e \u003cem\u003eKaddid\u003c/em\u003e or \u003cem\u003eEl-Guedid\u003c/em\u003e is a traditional meat product prepared all over the Algerian territory from sheep and beef meat whereas goat and camel meat is mainly used in sub-dry areas. Usually, the raw meat is cut into strips, seasoned abundantly with salt and spices and brine or dry salting was applied before dried either under the sun or in the shade [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. Southwestern Algerian \u003cem\u003eKaddid\u003c/em\u003e is mostly produced by adding salt and spices onto fresh lamb meat strips that are then hung to dry in the shade [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eAs part of \u003cem\u003eKaddid\u003c/em\u003e microbiota, LAB were reported to mainly involve species adapted to high NaCl concentration as \u003cem\u003eLactiplantibacillus (Lpb.) plantarum, Latilactobacillus (Lat.) sakei\u003c/em\u003e and \u003cem\u003eLat. curvatus\u003c/em\u003e described for Tunisian and Algerian dry-salted meat and \u003cem\u003eEl-Guedid\u003c/em\u003e [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. During fermentation, lactobacilli produce organic acids causing a pH reduction, but can also produce exopolysaccharides (EPS) reported to negatively affect the texture of fermented dry and salted meat products [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. LAB can also produce biogenic amines by amino acid decarboxylation that accumulates during fermentation/ripening, which influence organoleptic properties and health when consumed in high concentrations [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. Lactobacilli have a biopreservative role in fermented meat products by producing, in addition to organic acids, antimicrobial compounds such as bacteriocins. These ribosomal produced peptides present a variable spectrum of antimicrobial activity, usually against related species such as \u003cem\u003eL. monocyotgenes\u003c/em\u003e. This bacterium can infect humans and animals as opportunistic foodborne pathogen causing serious illness and is frequently isolated from food and food-processing environments [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. Bacteriocinogenic lactobacilli as bioprotective cultures are potentially used as natural shelf-life safety and preventing the growth of foodborne pathogens [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. Another relevant safety aspect of LAB is the presence of antibiotic resistance genes in mobile genetic elements, such as plasmids and transposons. In fact, these elements can be transferred to other species, including pathogenic bacteria, during food manufacture or during the passage through the gastrointestinal tract. Indeed, this poses an additional risk due to the ready-to-eat nature of the fermented meat products and their potential to become important antibiotic resistance reservoirs [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. Particularly, the presence of tetracycline and erythromycin resistant lactobacilli in meat-associated bacteria was reported [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. To avoid the spread of antibiotics resistance genes that may be part of GRAS LAB genome, beneficial LAB to be used as starter cultures must be evaluated on a strain basis [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. Therefore, the aim of this study was the isolation, identification and characterization of LAB from laboratory-made \u003cem\u003eKaddid\u003c/em\u003e to select an autochthonous functional culture with a main focus to prevent spoilage and ensure product safety.\u003c/p\u003e"},{"header":"2. Material and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e\u003cem\u003e2.1. Kaddid preparation and sampling\u003c/em\u003e\u003c/h2\u003e \u003cp\u003eA leg lamb meat (~\u0026thinsp;3 kg) 24 h \u003cem\u003epost-mortem\u003c/em\u003e was purchased from a butcher\u0026rsquo;s shop in M\u0026eacute;d\u0026eacute;a, Algeria. \u003cem\u003eKaddid\u003c/em\u003e preparation was carried out according to South-Western Algerian method [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. Meat was aseptically skinned and deboned manually and thin (2\u0026ndash;3 cm thick) and long (20\u0026ndash;30 cm) stripes were cut following anatomical lines. The strips were then homogeneously rubbed with a mix of salt (200 g), black pepper (2 g) and cumin (2 g) while sugar (10 g) was added to boost LAB growth and placed in a ventilated oven (Memmert, Germany) at 22\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C and 75\u0026ndash;80% (RH%) for 15 days. Physicochemical and microbiological analyses were carried out during drying-fermentation at 0, 3, 7 and 15 days.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2. Physicochemical and microbiological characterization of Kaddid. Isolation of LAB\u003c/h2\u003e \u003cp\u003eTotal titratable acidity (TTA), pH, moisture (%RH), water activity (a\u003csub\u003eW\u003c/sub\u003e) and microbiological analysis of \u003cem\u003eKaddid\u003c/em\u003e samples were performed as previously described Boubakri et al. [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. For microbiological analysis, each sample (25 g) suspended in 225 mL of sterile tryptone-salt (tryptone 1 g/L; NaCl 0.85 g/L; Tween 80 1 mL/L) and homogenized (Stomacher 400, Seward, Worthing, UK) was serially diluted. Viable cells enumeration of total aerobic mesophilic bacteria (TAMB) on Plate Count Agar (Britania, Argentina) incubated aerobically (48 h, 30\u0026deg;C) was carried out. In addition, counts of total coliforms, staphylococci and the detection of \u003cem\u003eSalmonella\u003c/em\u003e were achieved as described by Boubakri et al. [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. LAB viable counts were performed using MRS agar (Merck, Germany) microaerobically incubated at 28\u0026deg;C for 4 days. LAB isolation was carried out after 15 days of \u003cem\u003eKaddid\u003c/em\u003e ripening by randomly picking forty (40) single colonies from MRS plates and after purification by successive streaking, Gram and catalase tests were performed.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3. Identification of lactic acid bacteria\u003c/h2\u003e \u003cp\u003eLAB were preliminary grouped by their physiological and biochemical traits as previously described Boubakri et al. [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. Growth at 10 \u0026ordm;C and 45 \u0026ordm;C during 7 d and 24 h, respectively, at pH 4 to 9.6 and in the presence of 4 to 10% NaCl was evaluated in MRS (bacilli) and M17 (cocci) media. Production of gas from glucose and arginine hydrolysis were also examined. For molecular identification, genomic DNA was extracted according to Pospiech \u0026amp; Neumann [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. Strain differentiation was performed by repetitive sequence-based (rep-PCR) fingerprinting by using a (GTG)5 primer [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. The master mix contained 4 \u0026micro;L of 5x buffer (InbioHighway, Argentina), 2 \u0026micro;L of 5mM dNTPs (Promega, Madison, WI, USA), 2 U of Taq polymerase (InbioHighway, Argentina), 1 \u0026micro;L of 50 ng DNA template, 2 \u0026micro;L of 10 mM primer (GTG)5 (Sigma-Merck, Germany), and 4 \u0026micro;L of 25 mM MgCl\u003csub\u003e2\u003c/sub\u003e. Polymerase chain reactions consisted of an initial denaturation at 94\u0026deg;C for 5 min, 30-cycle denaturation reaction at 94\u0026deg;C for 1 min, 1-min annealing at 40\u0026deg;C, 8-min extension at 65\u0026deg;C, and a final extension at 65\u0026deg;C for 10 min. DNA quantification was performed by using the NANO UV/Vis Spectrophotometer (MicroDigital Co., South Korea). Amplification reactions were carried out in a MyCycler thermal cycler (Bio-Rad, Richmond, CA, USA). PCR-products were separated by electrophoresis on a 1.5% (w/v) agarose at 40 V for 190 min and the molecular size marker used was 1 kb Plus DNA ladder (Waltham, Massachusetts, USA). Gels were stained with GelRed (Biotium Inc., CA, USA) and visualized with an ultraviolet light transilluminator (320 nm) using Image Scanner III (GE, Health Care, Pittsburgh, USA). Genomic DNA of selected isolates in each cluster was used for amplification of the almost full-length 16S rRNA gene fragment (~\u0026thinsp;500pb) using the primers MLB and PLB [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e] and sequenced at CERELA-CONICET by an ABI 3130 DNA sequencer (Applied Biosystems, CA, USA). rRNA gene sequence alignments were performed using the multiple sequence alignment method and identification queries were fulfilled by a BLASTn search in GenBank (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://www.ncbi.nlm.nih.gov/GenBank/\u003c/span\u003e\u003cspan address=\"http://www.ncbi.nlm.nih.gov/GenBank/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e accessed on 20 July 2023).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e2.4. LAB Spoilage characteristics\u003c/h2\u003e \u003cdiv id=\"Sec7\" class=\"Section3\"\u003e \u003ch2\u003e2.4.1. Exopolysaccharides (EPS) production\u003c/h2\u003e \u003cp\u003eLAB ropiness was examined by the presence of a ropy condition after touching the colony with a loop as described by Ruiz Rodr\u0026iacute;guez et al. [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. EPS from glucose and sucrose (4%) were investigated. EPS quantification was carried out by inoculation of 18 h subcultures (1%) into 50 mL modified MRS medium containing sucrose (100 g/L). After incubation (30\u0026deg;C for 72 h), the cultures were centrifugated (6000 g for 30 min) and the supernatants were collected. The extraction of EPS was performed according to the technique of van Geel-Schutten et al. [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e] with minor modifications. Each supernatant was mixed with two-fold volume of cold ethanol (4\u0026deg;C). The supernatant-ethanol mixtures were kept for one night at 4\u0026deg;C and then centrifuged (2000 g for 15 min) to obtain EPS. The supernatants were removed, and EPS were subjected again to the same operation. Repeated rinsing of the precipitates was performed with absolute ethanol to eliminate any culture medium residues before drying at room temperature. The dried samples were diluted with ultrapure water and EPS levels were determined as mg glucose/L by phenol\u0026ndash;sulfuric acid method [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. Briefly, dried EPS was dissolved in deionized water until a final concentration of 5 mg/mL. To each 2 mL of EPS suspension, 50 \u0026micro;l of 80% (w/v) phenol and 5 mL of sulfuric acid (H\u003csub\u003e2\u003c/sub\u003eSO\u003csub\u003e4\u003c/sub\u003e) was added quickly. The sample was incubated at room temperature for 45 min and the optical density at 490 nm was measured using a 96 U-well polystyrene microtitre (MtPs) plates (ExtraGene, Taiwan) and a microplate reader (SAFAS MP 96, Monaco) equipped with SAFAS software. The standard curve with linear regression of L-glucose (Sigma-Aldrich) at OD\u003csub\u003e490\u003c/sub\u003e was performed with Microsoft Excel 10.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section3\"\u003e \u003ch2\u003e2.4.2. Biogenic amine production\u003c/h2\u003e \u003cp\u003eDecarboxylase medium plates containing different precursor amino acids at a 1% final concentration (L-histidine monohydrochloride, tyrosine disodium salt, L-ornithine monohydrochloride and lysine monohydrochloride from Sigma-Aldrich, Germany) were used to test biogenic amines production according to Bover-Cid \u0026amp; Holzapfel [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003e2.5. LAB Inhibitory activity\u003c/h2\u003e \u003cdiv id=\"Sec10\" class=\"Section3\"\u003e \u003ch2\u003e2.5.1. Antibacterial activity\u003c/h2\u003e \u003cp\u003eA semi-quantitative agar-spot-test was used to evaluate antibacterial activity of LAB isolates according to Fontana et al. [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. In this study, native cell free supernatants (native-CFS) obtained by centrifugation (7000\u0026times;g for 5min) were assayed against Gram-positive and Gram-negative bacteria used as target indicators (Table \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e). Positive antimicrobial activity of LABs native-CFS were neutralized with 4N NaOH (Ciccarelli, Argentina) and treated with catalase (1000 U/ml) (Sigma-Aldrich, USA) to obtain treated-CFS.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section3\"\u003e \u003ch2\u003e2.5.2. Characterization of bacteriocin-like inhibitory substance (BLIS) against L. monocytogenes FBUNT\u003c/h2\u003e \u003cp\u003eTo ascertain inhibitory activity, serial double dilutions of the inhibitory-treated cell-free supernatant (treated-CFS) were meticulously prepared using sterile distilled water. The agar-spot-test technique was subsequently employed for the determination of inhibitory potential [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. After incubation, plates were examined for the appearance of clear zones, and inhibitory titers (AU/mL) were calculated according to the following formula: T\u0026thinsp;=\u0026thinsp;1/V\u0026times;D, where T is the inhibitory titer in AU/mL, V is the volume of the spotted supernatant (5 \u0026micro;l), and D is the maximum dilution factor with activity (halo presence). In addition, to confirm the proteinaceous nature of inhibitory substance/s each collected treated-CFS proteolytic enzymes were added including proteinase K, protease II, protease XIV, trypsin, pepsin and lysozyme (all from Sigma Aldrich, USA) at a final concentration of 0.1 mg/mL, incubated for 1 h at 37 \u0026ordm;C, heated at 90 \u0026ordm;C for 5 min to halt enzymatic reactions and the inhibitory activity was determined by the agar-spot-test method. \u003cem\u003eLat. curvatus\u003c/em\u003e CRL705 [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e] and native-CFS were used as positive control.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section3\"\u003e \u003ch2\u003e2.5.3. Determination of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against L. monocytogenes FBUNT\u003c/h2\u003e \u003cp\u003eMIC was assessed in 96 U-well polystyrene microtitre (MtPs) plates (ExtraGene, Taiwan) as described by Ben Slama et al. [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. LAB treated-CFS were subjected to serial dilutions in Brain Heart Infusion broth (BHI) (Britania, Argentina) to yield a final concentration ranging from 5 to 90% of 200 \u0026micro;L total volume in each well, then 10 \u0026micro;L of \u003cem\u003eL. monocytogenes\u003c/em\u003e FBUNT at \u0026sim;10\u003csup\u003e9\u003c/sup\u003e CFU/mL were added to each well. MIC value was defined as the lowest concentration of treated-CFS without macroscopically visible growth. For the MBC determination, 10 \u0026micro;L of each well with no visible growth was poured on BHI agar plates and incubated at 30\u0026deg;C for 24 h. MBC was defined as the lowest concentration at which 99% of the bacteria were killed.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section3\"\u003e \u003ch2\u003e2.5.4. PCR detection of bacteriocin genes\u003c/h2\u003e \u003cp\u003eIn this study, genomic DNA of positive control and LAB strains primers including specific and degenerate primers (Table \u003cspan refid=\"MOESM2\" class=\"InternalRef\"\u003eS2\u003c/span\u003e) were designed and synthesized by Genbiotech (Buenos Aires, Argentina). PCR-amplifications were performed in 25 \u0026micro;l reaction mixture as described by Mechoud et al. [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. PCR-products were separated by electrophoresis on 1% (w/v) agarose at 80 V for 45 min. DNA quantification and stained gels were visualized as previously stated. The resulting PCR of the strain \u003cem\u003eLat. sakei\u003c/em\u003e subsp. \u003cem\u003esakei\u003c/em\u003e 26 of presumed \u003cem\u003esak\u003c/em\u003eG product was purified with Wizard\u0026reg; SV Gel and PCR Clean-Up system (Promega Corporation, USA) according to the manufacturer\u0026rsquo;s instructions. DNA was sequenced at CERELA-CONICET through an ABI 3130 DNA sequencer using the forward or reverse primer. Analysis of DNA sequences was performed with the BLASTn search in GenBank program available at \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://www.ncbi.nlm.nih.gov/GenBank/\u003c/span\u003e\u003cspan address=\"http://www.ncbi.nlm.nih.gov/GenBank/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e accessed on 26 October 2023.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section3\"\u003e \u003ch2\u003e2.5.5. LAB antifungal activity\u003c/h2\u003e \u003cp\u003eThe modified agar diffusion assay method described by Magnusson et al. [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e] was used against autochthonous molds and yeast (Table \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003e2.6. LAB antibiotic resistance/susceptibility\u003c/h2\u003e \u003cdiv id=\"Sec16\" class=\"Section3\"\u003e \u003ch2\u003e2.6.1. Phenotypic antibiotic resistance and minimum inhibitory concentration (MIC) determination\u003c/h2\u003e \u003cp\u003eMinimum inhibitory concentrations (mg/mL) of the antibiotics ampicillin (AMP), clindamycin (CLI), chloramphenicol (CHL), erythromycin (ERY), gentamicin (GEN), kanamycin (KAN), tetracycline (TET), streptomycin (STR) and vancomycin (Van) were determined for LAB strains according to the ISO 10932:2010 standard. Epidemiological cut-off values based on the recommendation of the European Committee on Antimicrobial Susceptibility Testing (EUCAST) and EFSA-FEEDAP [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e] were used. All antibiotics were purchased from Sigma-Aldrich (Missouri, USA) and ICN Biomedicals (California, USA). Each antibiotic was dissolved and filter-sterilized prior to its addition to mixed broth medium composed with 90% of ISM (Iso-Sensitest medium) from Oxoid (Thermofisher, Argentina) and 10% of MRS broth (Merck, Germany). The MIC was determined using the microdilution method described by the ISO/International Dairy Federation (IDF). The accuracy of susceptibility testing was monitored using quality control strain (\u003cem\u003eLpb. plantarum\u003c/em\u003e ATCC14917) and two isolated strains (\u003cem\u003eE. faecium\u003c/em\u003e K15 and K19).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec17\" class=\"Section3\"\u003e \u003ch2\u003e\u003cem\u003e2.6.2. PCR detection of antimicrobial resistance genes\u003c/em\u003e.\u003c/h2\u003e \u003cp\u003eThe presence of genes coding for antibiotic resistance (AR) in LAB strains phenotypically susceptible to antibiotics were evaluated through PCR reactions with the same reagents used above. The specific primers used, their target genes, the amplicon sizes, and the PCR protocol references used for gene detection are shown in Table \u003cspan refid=\"MOESM3\" class=\"InternalRef\"\u003eS3\u003c/span\u003e. The PCR amplifications were performed from total bacteria DNA obtained according to Pospiech \u0026amp; Neumann [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. The PCR products were separated, gel staining and visualization were performed as explained above.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec18\" class=\"Section2\"\u003e \u003ch2\u003e2.7. Statistical analysis\u003c/h2\u003e \u003cp\u003eAll \u003cem\u003ein-vitro\u003c/em\u003e assays were performed at least in duplicate with appropriate technical repetitions. Physicochemical and microbiological results were expressed as the mean value\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation of the data. Statistical analyses were performed with GraphPad Prism 7.04 Software using one-way ANOVA (nonparametric) followed by \u003cem\u003eTukey\u003c/em\u003e tests multiple comparison were \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05 was considered as significatively different unless otherwise indicated.\u003c/p\u003e \u003c/div\u003e"},{"header":"3. Results","content":"\u003cdiv id=\"Sec20\" class=\"Section2\"\u003e \u003ch2\u003e3.1. Physicochemical and microbial changes during the dry fermentation of Kaddid\u003c/h2\u003e \u003cp\u003eThe evolution of physicochemical parameters and microbial population during the drying-fermentation of \u003cem\u003eKaddid\u003c/em\u003e prepared under laboratory conditions is shown in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. LAB growth reached maximal numbers between 3 and 7 days correlating with the lower pH values; the accumulation of acids (% of lactic acid) kept increasing, reaching ΔTTA of 1.8%\u0026plusmn;0.2 at the end of fermentation in agreement with the previously reported for traditional salted \u003cem\u003eKaddid\u003c/em\u003e [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. In addition, it was observed that LAB and staphylococci are major populations in \u003cem\u003eKaddid\u003c/em\u003e, while coliforms were present in the meat (day 0) and then a remarkable reduction to undetectable level was observed.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ePhysicochemical and microbial (log CFU/g) changes during \u003cem\u003eKaddid\u003c/em\u003e dry-fermentation\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colspan=\"4\" nameend=\"c5\" namest=\"c2\"\u003e \u003cp\u003eSampling (days)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e15\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003epH\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e5.58\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5.18\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5.28\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e5.45\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eTTA (%)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.40\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.60\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.20\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eMoisture (%)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e67.41\u0026thinsp;\u0026plusmn;\u0026thinsp;0.42\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e32.21\u0026thinsp;\u0026plusmn;\u0026thinsp;3.12\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e25.71\u0026thinsp;\u0026plusmn;\u0026thinsp;3.58\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e17.89\u0026thinsp;\u0026plusmn;\u0026thinsp;1.83\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003ea\u003c/b\u003e\u003csub\u003e\u003cb\u003ew\u003c/b\u003e\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.87\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.79\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.71\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.58\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eTAMB\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4.45\u0026thinsp;\u0026plusmn;\u0026thinsp;0.77 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.50\u0026thinsp;\u0026plusmn;\u0026thinsp;0.54\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4.34\u0026thinsp;\u0026plusmn;\u0026thinsp;0.41\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4.36\u0026thinsp;\u0026plusmn;\u0026thinsp;0.25\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eLAB\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.71\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.53\u0026thinsp;\u0026plusmn;\u0026thinsp;0.39\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.40\u0026thinsp;\u0026plusmn;\u0026thinsp;0.33\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.22\u0026thinsp;\u0026plusmn;\u0026thinsp;0.17\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eStaphylococci\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.58\u0026thinsp;\u0026plusmn;\u0026thinsp;0.37\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.35\u0026thinsp;\u0026plusmn;\u0026thinsp;0.49\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.29\u0026thinsp;\u0026plusmn;\u0026thinsp;0.27\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.71\u0026thinsp;\u0026plusmn;\u0026thinsp;0.21\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eYeast \u0026amp; molds\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.43\u0026thinsp;\u0026plusmn;\u0026thinsp;0.27\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.24\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.15\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;1.00\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eTotal coliforms\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.73\u0026thinsp;\u0026plusmn;\u0026thinsp;0.41\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026lt; DL\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026lt; DL\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026lt; DL\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eSalmonella\u003c/b\u003e \u003cb\u003espp.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eND\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eND\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eND\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eND\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"5\"\u003eTTA: total titratable acidity; a\u003csub\u003eW\u003c/sub\u003e: water activity; TAMB: total aerobic mesophilic bacteria; LAB: lactic acid bacteria. Mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation; \u0026lt; DL: below the detection limit of the analytical method (10 CFU/g); ND: not detected (absence in 25 g). \u003csup\u003ea\u0026minus;d\u003c/sup\u003e: According to One-way ANOVA followed by \u003cem\u003eTukey\u003c/em\u003e test multiple comparison test (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05) with 0.001 (99.9% confidence interval), means in the row sharing similar letters are not significatively different (ns: 0.1234), whereas group superscript different letters differ significantly.\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec21\" class=\"Section2\"\u003e \u003ch2\u003e3.2. Identification of LAB\u003c/h2\u003e \u003cp\u003eBacteria isolates from the final stage of \u003cem\u003eKaddid\u003c/em\u003e drying-fermentation were investigated. Firstly, LAB isolates (29 bacilli/coccobacilli and 11 cocci) were subjected to a physiological preliminary identification (Table \u003cspan refid=\"MOESM4\" class=\"InternalRef\"\u003eS4\u003c/span\u003e). The bacilli/coccobacilli (71.79%) cluster involved aerobic and facultative heterofermenter isolates that would be assigned to lactobacilli, \u003cem\u003eBacillus\u003c/em\u003e or \u003cem\u003eWeissella\u003c/em\u003e genera with variable arginine hydrolysis, growth ability between 10 and 45\u0026deg;C in a wide range of pH except for pH 9.6 and resistant to high NaCl concentrations. Whereas homofermenter tetrads- and chain-forming cocci (10.25%) cluster growing between 10 to 45\u0026deg;C, up to 9.6% NaCl but not at pH\u0026thinsp;\u0026gt;\u0026thinsp;8.0 were presumed as \u003cem\u003ePediococcus\u003c/em\u003e. However, the cluster of chain-forming homofermentative cocci (17.94%) developing at the same temperatures, in a wider pH range (4.0 to 9.6) and up to 6.5% NaCl were assigned to \u003cem\u003eEnterococcus\u003c/em\u003e genus. Isolates were then sub-cultured (one single isolate from clustering profile of heterofermentative bacilli failed to grow and was discarded). Thus, a total of 39 strains showing growth were identified by (GTG)5-PCR and 16S rRNA sequencing yielding to a variable bands of molecular size ranging from 300 to 4000 bp corresponding to the genera \u003cem\u003eLactiplantibacillus\u003c/em\u003e, \u003cem\u003eLatilactobacillus\u003c/em\u003e, \u003cem\u003ePediococcus\u003c/em\u003e, \u003cem\u003eEnterococcus\u003c/em\u003e and \u003cem\u003eWeissella\u003c/em\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Ascription of \u003cem\u003eKaddid\u003c/em\u003e isolates into species was based on the clusters derived from (GTG)5-PCR analysis; strains showing identical rep-PCR band patterns were considered as one rep-PCR biotype. Acordinly, isolates were grouped to 10 different (GTG)5-PCR biotypes. At least one representative from each biotype was identified by partial 16S rRNA gene sequencing. Biotype information from (GTG)5-PCR for LAB isolates (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e) allowed the association with \u003cem\u003eLactiplantibacillus (Lpb.) paraplantarum\u003c/em\u003e (K) 1, 2, 3, 4, 7, 8, 10, 12, 13, 14, 16, 17, 18, 20, 21, 31; \u003cem\u003eLatilactobacillus (Lat.) sakei\u003c/em\u003e (K) 5, 25; \u003cem\u003eLat. sakei subsp. sakei\u003c/em\u003e (K) 22, 23, 24, 26, 27, 28, 29, 30; \u003cem\u003ePediococcus (P.) acidilactici\u003c/em\u003e (K) 34, 36, 37, 38; \u003cem\u003eEnterococcus (E.) faecium\u003c/em\u003e (K) 6, 9, 11, 15, 19; \u003cem\u003eE. hirae\u003c/em\u003e (K) 32, 33, \u003cem\u003eWeissella (W.) cibaria\u003c/em\u003e (K) 35, 39 and \u003cem\u003eListeria (L.) monocytogenes\u003c/em\u003e FBUNT (40) used as indicator strain for antibacterial survey.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec22\" class=\"Section2\"\u003e \u003ch2\u003e3.3. Screening of antimicrobial activity of LAB\u003c/h2\u003e \u003cp\u003eInhibitory ability of LAB isolated from \u003cem\u003eKaddid\u003c/em\u003e was evaluated using Gram-positive/Gram-negative bacteria and yeast/molds as indicators (Table \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e). High antagonistic activity against Gram positive indicator strains was found (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Inhibitory activity of LAB native-CFS was observed for \u003cem\u003eLpb. paraplantarum Lat. sakei\u003c/em\u003e/subsp. \u003cem\u003esakei\u003c/em\u003e and \u003cem\u003eE. faecium\u003c/em\u003e against \u003cem\u003eL. monocytogenes\u003c/em\u003e FBUNT, \u003cem\u003eL. innocua\u003c/em\u003e CLIP74915\u003csup\u003eT\u003c/sup\u003e, \u003cem\u003eE. faecalis\u003c/em\u003e ATCC29212 and \u003cem\u003eB. subtilis\u003c/em\u003e ATCC13453, whereas a lack of inhibitory activity was observed against Gram negative bacteria (data not shown). When LAB native-CFS were neutralized and treated with catalase (treated-CFS), several LAB strains maintained their inhibitory activity against \u003cem\u003eListeria\u003c/em\u003e; titration of bacteriocin-like inhibitory substance (BLIS) showed a moderate antagonistic activity ranged between 200 and 400 AU/ml compared to the positive control \u003cem\u003eLat. curvatus\u003c/em\u003e CRL705 displaying an activity of 800 AU/ml. Moreover, antifungal activity against meat-borne molds showed most of LAB exhibiting a strong antagonism against \u003cem\u003ePenicillium\u003c/em\u003e and \u003cem\u003eCryptococcus\u003c/em\u003e (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). The highest antifungal activity was produced by \u003cem\u003eLpb. paraplantarum\u003c/em\u003e K1, K7, K13, K16, K17, K18 \u003cem\u003eLat. sakei\u003c/em\u003e K25, \u003cem\u003eLat. sakei\u003c/em\u003e subsp. \u003cem\u003esakei\u003c/em\u003e K23, K24, K28 as well as \u003cem\u003eE. faecium\u003c/em\u003e K6, K11, K19 strains. However, assayed LAB failed to inhibit the growth of \u003cem\u003eAspergillus\u003c/em\u003e, \u003cem\u003eAlternaria\u003c/em\u003e fungus and the commercial \u003cem\u003eSaccharomyces cerevisiae\u003c/em\u003e strain. When antifungal compounds were investigated on LAB treated-CFS, a loss of LAB inhibitory activity was found, indicating that major antifungal compounds in CFS would be organic acids.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eFrequency of antibacterial activity of LAB isolated from \u003cem\u003eKaddid\u003c/em\u003e against target indicators *BLIS: Bacteriocin\u0026ndash;like inhibitory substance; **Positive controls of bacteriocin production. No inhibition detection (\u0026mdash;); No determined (ND). Target strains were listed in table \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"11\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c11\" colnum=\"11\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eIdentified LAB from Kaddid\u003c/p\u003e \u003cp\u003e(\u003cem\u003en\u003c/em\u003e=39)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"7\" nameend=\"c8\" namest=\"c2\"\u003e \u003cp\u003eAntibacterial activity of LAB native-CFS against sensitive indicators\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"3\" nameend=\"c11\" namest=\"c9\"\u003e \u003cp\u003eBLIS* titration\u003c/p\u003e \u003cp\u003eagainst \u003cem\u003eL. monocytogenes\u003c/em\u003e FBUNT (AU/mL)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eL. monocytogenes\u003c/em\u003e FBUNT\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eL. innocua\u003c/em\u003e\u003c/p\u003e \u003cp\u003eL1PE\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e\u003cem\u003eL.\u003c/em\u003e\u003c/p\u003e \u003cp\u003e\u003cem\u003einnocua\u003c/em\u003e\u003c/p\u003e \u003cp\u003eCLIP74915\u003csup\u003eT\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cem\u003eS. aureus\u003c/em\u003e ATCC29213\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cem\u003eE. faecalis\u003c/em\u003e ATCC29212\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cem\u003eB. subtilis\u003c/em\u003e ATCC13453\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003e200\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c10\"\u003e \u003cp\u003e400\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c11\"\u003e \u003cp\u003e800\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eLpb. paraplantarum\u003c/b\u003e\u003c/p\u003e \u003cp\u003e(16 strains)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e83.30%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e61.05%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e5.55%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e61.05%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e49.95%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eK7, K8, K16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eK18, K21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eLat. sakei subsp. sakei\u003c/b\u003e\u003c/p\u003e \u003cp\u003e(8 strains)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003e12.50%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eK23, K24, K27, K28, K29, K30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eK22, K26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eLat. sakei\u003c/b\u003e\u003c/p\u003e \u003cp\u003e(2 strains)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e50%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e50%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e50%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e50%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eK25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eP. acidilactici\u003c/b\u003e\u003c/p\u003e \u003cp\u003e(4 strains)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e75%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e25%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"3\" nameend=\"c11\" namest=\"c9\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eE. faecium\u003c/b\u003e\u003c/p\u003e \u003cp\u003e(5 strains)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e60%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e60%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e60%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eK6, K9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eK11, K15, K19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eE. hirae\u003c/b\u003e\u003c/p\u003e \u003cp\u003e(2 strains\u003cb\u003e)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"3\" nameend=\"c11\" namest=\"c9\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eW. cibaria\u003c/b\u003e\u003c/p\u003e \u003cp\u003e(2 strains)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e50%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"3\" nameend=\"c11\" namest=\"c9\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eLat. curvatus\u003c/b\u003e \u003cb\u003eCRL705**\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eND\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eND\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eND\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eND\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"3\" nameend=\"c11\" namest=\"c9\"\u003e \u003cp\u003e800\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eLat. curvatus\u003c/b\u003e \u003cb\u003eCRL1532**\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eND\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eND\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eND\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eND\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"3\" nameend=\"c11\" namest=\"c9\"\u003e \u003cp\u003eND\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eAntifungal activity of LAB isolated from \u003cem\u003eKaddid\u003c/em\u003e against sensitive target strains CFS: cell free supernatant; \u0026mdash; (no growth suppression), + (1\u0026ndash;5 mm), ++ (5\u0026ndash;10 mm), +++ (˃ 10 mm) of inhibition zone. Target strains were listed in table \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eIdentified LAB from Kaddid\u003c/p\u003e \u003cp\u003e(\u003cem\u003en\u003c/em\u003e=39)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"5\" nameend=\"c6\" namest=\"c2\"\u003e \u003cp\u003eInhibitory activity of LAB CFS against target indicators\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003ePenicillium\u003c/em\u003e S4J15\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003ePenicillium\u003c/em\u003e\u003c/p\u003e \u003cp\u003eC7J0\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003eRhizopus\u003c/em\u003e\u003c/p\u003e \u003cp\u003eC5J0\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003eCryptococcus\u003c/em\u003e S3J0\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cem\u003eCryptococcus\u003c/em\u003e\u003c/p\u003e \u003cp\u003eC3J7\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eLbp. paraplantarum\u003c/b\u003e\u003c/p\u003e \u003cp\u003e(16 strains)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eK1 (+++)\u003c/p\u003e \u003cp\u003eK7 (++)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eK1, K7 (++)\u003c/p\u003e \u003cp\u003eK13, K16, K17 K18 (+)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eK1, K7 (++)\u003c/p\u003e \u003cp\u003eK16, K17, K18 (+)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eK1 (+++)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eK1, K7, K13, K16, K17, K18 (+++)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eLat. sakei subsp. sakei\u003c/b\u003e (8 strains)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eK28 (++)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eK23 (+++)\u003c/p\u003e \u003cp\u003eK24 (++), K28 (+)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eK23, K24, K28 (+++)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eK23, K24, K28 (+++)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eLat. sakei\u003c/b\u003e\u003c/p\u003e \u003cp\u003e(2 strains)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eK25 (+)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eK25 (++)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eK25 (++)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eK25 (+++)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eP. acidilactici\u003c/b\u003e\u003c/p\u003e \u003cp\u003e(4 strains)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eE. faecium\u003c/b\u003e\u003c/p\u003e \u003cp\u003e(5 strains)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eK6, K11, K19 (+++)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eK6, K11 (+++)\u003c/p\u003e \u003cp\u003eK19 (+)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eK6, K11, K19 (+)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eK6, K11, K19 (+++)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eK6, K11, K19 (+++)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eE. hirae\u003c/b\u003e\u003c/p\u003e \u003cp\u003e(2 strains)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eW. cibaria\u003c/b\u003e\u003c/p\u003e \u003cp\u003e(2 strains)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eK35 (++)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cdiv id=\"Sec23\" class=\"Section3\"\u003e \u003ch2\u003e3.3.1. Characterization of BLIS against L. monocytogenes FBUNT\u003c/h2\u003e \u003cp\u003eThe different interference patterns on the inhibitory activity of tested BLIS from \u003cem\u003eKaddid\u003c/em\u003e lactobacilli are shown in Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e. When the effect of temperature and pH on lactobacilli BLIS was evaluated, a loss of inhibitory activity after treatments at 60 and 80\u0026deg;C was observed, while residual activity (200 AU/mL) after treatment at 100\u0026deg;C/30 min was found for \u003cem\u003eLat. sakei\u003c/em\u003e K25 and \u003cem\u003eLat. sakei\u003c/em\u003e subsp. \u003cem\u003esakei\u003c/em\u003e K26 and after 121\u0026deg;C/15 min for \u003cem\u003eLpb. paraplantarum\u003c/em\u003e K21. Even lower, all tested strains preserved their antibacterial activity after storage at 5\u0026deg;C for 7 d. In addition, highest activity (400 AU/mL) was shown at pH 6.5 that was retained up to pH 4.5 (200 AU/mL), but a lack of inhibition was found at pHs 2, 8 and 10. Results indicated that BLIS from \u003cem\u003eLpb. paraplantarum\u003c/em\u003e K18 and K21, \u003cem\u003eLat. sakei\u003c/em\u003e K25 and \u003cem\u003eLat. sakei\u003c/em\u003e subsp. \u003cem\u003esakei\u003c/em\u003e K26 strains were inactivated by protease XIV and proteinase K; protease V was effective at inactivating BLIS from \u003cem\u003eLat. sakei\u003c/em\u003e K25 and \u003cem\u003eLat. sakei\u003c/em\u003e subsp. \u003cem\u003esakei\u003c/em\u003e K26, whereas lysosyme, pepsine and trypsine were unable to inactivate BLIS activity of any of the assayed lactobacilli (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). Additionally, microdilution method used to determine the MIC of BLIS against \u003cem\u003eL. monocytogenes\u003c/em\u003e FBUNT, showed 20% (80 AU/mL) of BLIS as effective for all tested strains with variable MBC (between 70 and 80%); a lower inhibitory potency of \u003cem\u003eKaddid\u003c/em\u003e strains (4xMIC) compared to \u003cem\u003eLat. curvatus\u003c/em\u003e CRL705 (MBC\u0026gt;100%) was detected. Furthermore, the presence of bacteriocin encoding genes was also evaluated (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e; Fig. \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e). The results revealed that strains \u003cem\u003eLpb. paraplantarum\u003c/em\u003e K18 and \u003cem\u003eLat. sakei\u003c/em\u003e subsp. \u003cem\u003esakei\u003c/em\u003e K26 exhibited comparable genetic profiles for bacteriocin-related genes; although both strains demonstrated the presence of amplified products for \u003cem\u003eskg\u003c/em\u003eA (sakacin G) and \u003cem\u003esppA\u003c/em\u003e (sakacin P); \u003cem\u003eLat. sakei\u003c/em\u003e K25 and \u003cem\u003eLat. sakei\u003c/em\u003e subsp. \u003cem\u003esakei\u003c/em\u003e K22 showed positive results only for \u003cem\u003eskg\u003c/em\u003eA and \u003cem\u003espp\u003c/em\u003eA respectively. Structural genes for curvacin A (\u003cem\u003esap\u003c/em\u003eA) and sakacin Q (\u003cem\u003espp\u003c/em\u003eQ) were not present in the tested lactobacilli from \u003cem\u003eKaddid\u003c/em\u003e and \u003cem\u003eLpb. paraplantarum\u003c/em\u003e K21 lack the encoding genes for sakacin.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eBLIS activity characterization and presence of bacteriocin genes in selected lactobacilli from \u003cem\u003eKaddid\u003c/em\u003e\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e\u003cem\u003eLpb. paraplantarum\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003eLat. sakei\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e \u003cp\u003e\u003cem\u003eLat. sakei\u003c/em\u003e subsp. \u003cem\u003esakei\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e\u003cem\u003eLat. curvatus\u003c/em\u003e\u003c/p\u003e \u003cp\u003eCRL705 (control)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003eK18\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003eK21\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003eK25\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003eK22\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003eK26\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eControl\u003c/b\u003e\u003c/p\u003e \u003cp\u003eMRS, pH 6.5/30\u0026deg;C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e400\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e400\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e400\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e400\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e400\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e800\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eTemperature*\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e45\u0026deg;C/60 min\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e400\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e400\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e400\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e400\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e400\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e600\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e60\u0026deg;C/60 min\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e600\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e80\u0026deg;C/30 min\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e400\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e100\u0026deg;C/30 min\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e200\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e200\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e200\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e121\u0026deg;C/15 min\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e200\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e200\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e200\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e200\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e5\u0026deg;C/ 7 d\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e200\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e200\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e200\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e200\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e200\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e200\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003epH*\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e4.5/30 \u0026ordm;C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e200\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e200\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e200\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e200\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e200\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e400\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2, 8, 10/30 \u0026ordm;C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eEnzymes **\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLysozyme/pepsine/trypsine\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eprotease V\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eprotease XIV\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eproteinase K\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eMIC (%)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e20\u003c/p\u003e \u003cp\u003e(80 AU/mL)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e20\u003c/p\u003e \u003cp\u003e(80 AU/mL)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e20\u003c/p\u003e \u003cp\u003e(80 AU/mL)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e20\u003c/p\u003e \u003cp\u003e(80 AU/mL)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e20\u003c/p\u003e \u003cp\u003e(80 AU/mL)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e5\u003c/p\u003e \u003cp\u003e(40 AU/mL)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eMBC (%)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e70\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e70\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e70\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e˃100\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eBacteriocin gene(s\u003c/b\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003eskg\u003c/b\u003e\u003cb\u003eA\u003c/b\u003e, \u003cb\u003espp\u003c/b\u003e\u003cb\u003eA\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003eND\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003eskg\u003c/b\u003e\u003cb\u003eA\u003c/b\u003e,\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003espp\u003c/b\u003e\u003cb\u003eA\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003eskg\u003c/b\u003e\u003cb\u003eA\u003c/b\u003e, \u003cb\u003espp\u003c/b\u003e\u003cb\u003eA\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003espp\u003c/b\u003e\u003cb\u003eA\u003c/b\u003e, \u003cb\u003espp\u003c/b\u003e\u003cb\u003eQ\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"7\"\u003e*Results are indicated as AU/mL; ** (+): presence of inhibition halo (\u0026ge;\u0026thinsp;6 mm); (-) absence of inhibition halo. \u003cem\u003eLat. sakei\u003c/em\u003e CRL1862 and \u003cem\u003eLat. curvatus\u003c/em\u003e CRL1537 used as control for curvacin A (\u003cem\u003esap\u003c/em\u003eA) and sakacin P (\u003cem\u003espp\u003c/em\u003eA) genes (Fontana et al., 2015). and sakacin G (\u003cem\u003eskg\u003c/em\u003eA), this study.\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec24\" class=\"Section2\"\u003e \u003ch2\u003e3.4. Spoilage potential of Kaddid lactobacilli\u003c/h2\u003e \u003cp\u003eThe phenotypical detection of gas, exopolysacharides (EPS) and biogenic amines (BA) production by \u003cem\u003eKaddid\u003c/em\u003e selected strains were investigated (Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e). All the tested strains displayed gas production from gluconate, while only \u003cem\u003eLat. sakei\u003c/em\u003e subsp. \u003cem\u003esakei\u003c/em\u003e K22 produced minimal amount of gas from glucose in correlation with their facultative heterofermentative character. Even in lower abundance compared to positive control, a ropy phenotype from glucose for K25, K22 and K26 lactobacilli was detected while only K22 strain presented ropiness from sucrose. The ropiness phenotype for \u003cem\u003eLpb. paraplantarum\u003c/em\u003e K18 and K21 strains was barely detectable while that of \u003cem\u003eLat. sakei\u003c/em\u003e K25 and \u003cem\u003eLat. sakei\u003c/em\u003e subsp. sakei K22 and K26 were more detectable (Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e). Even EPS production values were lower than that from \u003cem\u003eW. cibaria\u003c/em\u003e BK19 (2464.62\u0026thinsp;\u0026plusmn;\u0026thinsp;113.34 mg/L) used as control, EPS production by K22 from sucrose exhibited the highest value of 574.62\u0026thinsp;\u0026plusmn;\u0026thinsp;2mg/L. No significant differences (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05) were found between the remaining strains with lower EPS production values. In addition, when \u003cem\u003eKaddid\u003c/em\u003e selected lactobacilli were tested for their decarboxylase ability, none of the assayed strains were observed to produce bioactive amines (data not shown).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab5\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eSpoilage potential of selected lactobacilli from \u003cem\u003eKaddid\u003c/em\u003e\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eLactobacilli\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eGas from\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003eRopiness from\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eEPS (as mg of glucose/L) *\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eglucose\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003egluconate\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eglucose\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003esucrose\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eLpb. paraplantarum\u003c/b\u003e \u003cb\u003eK18\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e138.37\u0026thinsp;\u0026plusmn;\u0026thinsp;05.36\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eLpb. paraplantarum\u003c/b\u003e \u003cb\u003eK21\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e327.12\u0026thinsp;\u0026plusmn;\u0026thinsp;22.54\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eLat. sakei\u003c/b\u003e \u003cb\u003eK25\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e258.37\u0026thinsp;\u0026plusmn;\u0026thinsp;11.48\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eLat. sakei\u003c/b\u003e \u003cb\u003esubsp\u003c/b\u003e. \u003cb\u003esakei\u003c/b\u003e \u003cb\u003eK22\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e574.62\u0026thinsp;\u0026plusmn;\u0026thinsp;24.97\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eLat. sakei\u003c/b\u003e \u003cb\u003esubsp\u003c/b\u003e. \u003cb\u003esakei\u003c/b\u003e \u003cb\u003eK26\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e452.12\u0026thinsp;\u0026plusmn;\u0026thinsp;23.88\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eLpb. plantarum\u003c/b\u003e \u003cb\u003eCRL972\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eND\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eND\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eND\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eLat. sakei\u003c/b\u003e \u003cb\u003eCRL1407\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eND\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eND\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eND\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eW. cibaria\u003c/b\u003e \u003cb\u003eBK19\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eND\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eND\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2464.62\u0026thinsp;\u0026plusmn;\u0026thinsp;113.34\u003csup\u003ef\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"6\"\u003ePositive test (+, ++, +++); negative test (-); not determined (ND). \u003cem\u003eLpb. plantarum\u003c/em\u003e CRL972 and \u003cem\u003eLat. sakei\u003c/em\u003e CRL1407 (CERELA culture collection) and \u003cem\u003eW. cibaria\u003c/em\u003e BK19 were used as positive control for gas and EPS production, respectively. *: EPS from 100g/L of sucrose; \u003csup\u003ea\u0026minus;f\u003c/sup\u003e: According to One-way ANOVA followed by \u003cem\u003eTukey\u003c/em\u003e test multiple comparison test (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05) with 0.001 (99.9% confidence interval), means in the same column sharing similar letters are not significatively different (ns: 0.1234), whereas group superscript different letters differ significantly.\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec25\" class=\"Section2\"\u003e \u003ch2\u003e3.5. Antibiotic resistance and detection of AR gene(s)\u003c/h2\u003e \u003cp\u003eThe analysis was performed following the cut-off value and the indications given by EFSA [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. The MICs determination according to ISO/IDF of eight antibiotics was examined in the selected \u003cem\u003eKaddid\u003c/em\u003e lactobacilli and two isolates of \u003cem\u003eE. faecium\u003c/em\u003e (K15 and K19), (Table\u0026nbsp;\u003cspan refid=\"Tab6\" class=\"InternalRef\"\u003e6\u003c/span\u003e). \u003cem\u003eKaddid\u003c/em\u003e lactobacilli revealed an unexpected high resistance to TET, a double resistance TET/STR for \u003cem\u003eLat. sakei\u003c/em\u003e K25, whereas \u003cem\u003eLpb. paraplantarum\u003c/em\u003e K18 was the only strain with no phenotypical antibiotic resistance and all lactobacilli were susceptible to the rest of the evaluated antibiotics. \u003cem\u003eLat. sakei\u003c/em\u003e K25 displayed MIC values for STR three-fold the cut-off value (64 \u0026micro;L/g), whereas a MIC of four-fold the cut-off value (8 \u0026micro;L/g) was found for TET. When the presence of AR gene(s) was investigated by PCR, \u003cem\u003etet\u003c/em\u003eM gene was found in all strains even in \u003cem\u003eLpb. paraplantarum\u003c/em\u003e K18 that was phenotypically sensitive to TET (Table\u0026nbsp;\u003cspan refid=\"Tab6\" class=\"InternalRef\"\u003e6\u003c/span\u003e; Fig. \u003cspan refid=\"MOESM2\" class=\"InternalRef\"\u003eS2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab6\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 6\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eMICs distribution and antibiotic resistance (AR) genes among \u003cem\u003eKaddid\u003c/em\u003e selected lactobacilli\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"15\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c11\" colnum=\"11\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c12\" colnum=\"12\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c13\" colnum=\"13\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c14\" colnum=\"14\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c15\" colnum=\"15\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eAMP\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e \u003cp\u003eCLI\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c8\" namest=\"c7\"\u003e \u003cp\u003eCHL\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003eERY\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c10\"\u003e \u003cp\u003eGEN\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c11\"\u003e \u003cp\u003eKAN\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c12\"\u003e \u003cp\u003eSTR\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c13\"\u003e \u003cp\u003eTET\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c14\"\u003e \u003cp\u003eVAN\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c15\"\u003e \u003cp\u003eAR genes\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colspan=\"14\" nameend=\"c14\" namest=\"c1\"\u003e \u003cp\u003e\u003cem\u003eLpb. paraplantarum\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c15\"\u003e\u0026nbsp;\u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e\u003cem\u003eCut-off value\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e\u003cem\u003e2\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e\u003cem\u003e4\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cem\u003e8\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u003cem\u003e1\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e\u003cem\u003e16\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e\u003cem\u003e64\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e\u003cem\u003en.r.\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e\u003cem\u003e32\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e\u003cem\u003en.r\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eK18\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eMIC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e\u0026lt;0.032\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e\u0026lt;0.032\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.125\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u0026lt;0.016\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e\u0026lt;0.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e\u0026gt;\u0026thinsp;128\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e\u003cb\u003etet\u003c/b\u003e\u003cb\u003eM\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eK21\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e0.125\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e\u0026lt;0.032\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.125\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u0026lt;0.016\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e0.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e128\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e\u003cb\u003e64\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e\u0026gt;\u0026thinsp;128\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e\u003cb\u003etet\u003c/b\u003e\u003cb\u003eM\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"14\" nameend=\"c14\" namest=\"c1\"\u003e \u003cp\u003e\u003cb\u003eLat. sakei\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" morerows=\"1\" nameend=\"c2\" namest=\"c1\" rowspan=\"2\"\u003e \u003cp\u003e\u003cb\u003eK25\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eCut-off value\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e\u003cem\u003e2\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e\u003cem\u003e4\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cem\u003e8\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u003cem\u003e1\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e\u003cem\u003e16\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e\u003cem\u003e64\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e\u003cem\u003e64\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e\u003cem\u003e32\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e\u003cem\u003en.r\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eMIC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e0.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e\u0026lt;0.032\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.125\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u0026lt;0.016\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e\u003cb\u003e256\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e\u003cb\u003e64\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e\u0026gt;\u0026thinsp;128\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e\u003cb\u003etet\u003c/b\u003e\u003cb\u003eM\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"14\" nameend=\"c14\" namest=\"c1\"\u003e \u003cp\u003e\u003cb\u003eLat. sakei\u003c/b\u003e \u003cb\u003esubsp\u003c/b\u003e. \u003cb\u003esakei\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003e\u003cb\u003eK22\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eMIC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e0.125\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e\u0026lt;0.032\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.125\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u0026lt;0.016\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e0.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e\u0026lt;2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e32\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e\u003cb\u003e64\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e\u0026gt;\u0026thinsp;128\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e\u003cb\u003etet\u003c/b\u003e\u003cb\u003eM\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003e\u003cb\u003eK26\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e0.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e\u0026lt;0.032\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.125\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u0026lt;0.016\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e\u0026lt;2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e32\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e\u003cb\u003e64\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e\u0026gt;\u0026thinsp;128\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e\u003cb\u003etet\u003c/b\u003e\u003cb\u003eM\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"14\" nameend=\"c14\" namest=\"c1\"\u003e \u003cp\u003e\u003cb\u003eE. faecium\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eCut-off value\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e\u003cem\u003e2\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e\u003cem\u003e4\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cem\u003e16\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u003cem\u003e4\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e\u003cem\u003e32\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e\u003cem\u003e1024\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e\u003cem\u003e128\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e\u003cem\u003e4\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003e\u003cb\u003eK15\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eMIC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e\u003cb\u003e4\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.125\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u003cb\u003e8\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e32\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e128\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e\u0026lt;0.125\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003e\u003cb\u003eK19\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e\u003cb\u003e4\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003e\u003cb\u003e16\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.125\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u003cb\u003e8\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e32\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e1024\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e\u003cb\u003e256\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e\u003cb\u003e64\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0,25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"15\"\u003e\u003cem\u003eCut-off values\u003c/em\u003e were proposed by the EFSA-FEEDAP (2018) and MIC are expressed in \u0026micro;g/mL; numbers in bold\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"15\"\u003eindicate antibiotic resistance; \u003cem\u003en.r\u003c/em\u003e: not required.\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"4. Discussion","content":"\u003cp\u003eDuring \u003cem\u003eKaddid\u003c/em\u003e production a lactic fermentation takes place as reported in other dry-cured and salted meat products such as \u0026ldquo;\u003cem\u003eLac\u0026oacute;n\u003c/em\u003e\u0026rdquo; [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. The small size and thickness of \u003cem\u003eKaddid\u003c/em\u003e strips represent a large surface-to-weight ratio favoring dehydration during post-salting and drying stages leading to great moisture loss and a\u003csub\u003ew\u003c/sub\u003e decrease. The presence of high salt concentration helps to draw water and sugars out of meat tissue; salt uptake and water exudation from meat are mutually dependent [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. On the other hand, microbiological traits at 15 days showed LAB and staphylococci as major populations in \u003cem\u003eKaddid\u003c/em\u003e. LAB presence was found to be higher than that reported for Tunisian \u003cem\u003eKaddid\u003c/em\u003e [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e], this correlates with the controlled conditions of laboratory-made \u003cem\u003eKaddid\u003c/em\u003e. The presence of coagulase-negative staphylococci agrees with the abundant presence of \u003cem\u003eStaphylococcus\u003c/em\u003e genus in salted/cured meat [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e], which are tolerant to high salt concentration. In accordance, several LAB species were reported to adapt to high NaCl concentration, particularly \u003cem\u003eLat. sakei\u003c/em\u003e that was identified from salted anchovies resisting up to 15% of NaCl [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]. The reduction of coliforms level was in correlation with the increased LAB growth and acid production, representing a good hygienic indicator after preparation/handling of meat for \u003cem\u003eKaddid\u003c/em\u003e production.\u003c/p\u003e \u003cp\u003eResults of LAB molecular identification agrees with those from high salt-containing meat products [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. In fact, the presence of the phylum Firmicutes, including \u003cem\u003eLactobacillaceae\u003c/em\u003e, \u003cem\u003eLeuconostocaceae\u003c/em\u003e and \u003cem\u003eEnterococcaceae\u003c/em\u003e families was widely reported from dry-salted meat and sea food products [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e, \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e, \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. The dominance of lactobacilli species at the final stage of the dry-ripening salted and/or cured dry-meat products is in correlation with their ability to adapt to high NaCl during fermentation, processing and storage. Indeed, \u003cem\u003eLat. sakei\u003c/em\u003e identification was reported for Algerian \u003cem\u003eEl-Guedid\u003c/em\u003e [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. Furthermore, \u003cem\u003eLat. sakei\u003c/em\u003e species was widely identified as predominant from many dry-fermented sausages across the Mediterranean region [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e, \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e, \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e, \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. The presence of \u003cem\u003eLat. sakei\u003c/em\u003e and \u003cem\u003eLat. sakei\u003c/em\u003e subsp. \u003cem\u003esakei\u003c/em\u003e in laboratory-made \u003cem\u003eKaddid\u003c/em\u003e could be assigned to the dry-ripening temperature (22\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C) used, in agreement with the detection of \u003cem\u003eLat. sakei\u003c/em\u003e and \u003cem\u003eLat. curvatus\u003c/em\u003e in sausages ripened at low temperatures while \u003cem\u003eLpb. plantarum\u003c/em\u003e was dominant at higher temperatures (\u0026gt;\u0026thinsp;25\u0026deg;C) as was reported by L\u0026uuml;cke et al. [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. The presence of \u003cem\u003eLpb. paraplantarum\u003c/em\u003e was also reported from Andalusian traditional spontaneous fermented sausages [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e], whereas \u003cem\u003eWeissella\u003c/em\u003e was isolated from traditional homemade \u003cem\u003eKaddid\u003c/em\u003e prepared under Saharan conditions in South-Western Algeria [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. In addition, added spices to raw meat may harbor a diverse and abundant microbial community with LAB species such as \u003cem\u003eWeissella, Pediococcus\u003c/em\u003e and \u003cem\u003eEnterococci\u003c/em\u003e [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIt is known that metabolic compounds produced by LAB, including organic acids, hydrogen peroxide and bacteriocins, can exert antimicrobial effect against a range of pathogens and contaminants [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e]. The highest inhibitory activity (400 AU/mL) after CFS titration against \u003cem\u003eL. monocytogenes\u003c/em\u003e FBUNT was observed for \u003cem\u003eLpb. paraplantarum\u003c/em\u003e K18 and K21, \u003cem\u003eLat. sakei\u003c/em\u003e K25, \u003cem\u003eLat. sakei\u003c/em\u003e subsp. \u003cem\u003esakei\u003c/em\u003e K22, K26 and \u003cem\u003eE. faecium\u003c/em\u003e K11, K15, K19 strains and may be attributed to bacteriocin production. When antifungal activity against meat-borne molds were analyzed, \u003cem\u003eLat. sakei\u003c/em\u003e and \u003cem\u003eLpb. paraplantarum\u003c/em\u003e strains showed high activity against \u003cem\u003ePenicillium\u003c/em\u003e in agreement with that of \u003cem\u003eLat. sakei\u003c/em\u003e subsp. ALI033 isolated from \u003cem\u003eKimchi\u003c/em\u003e which was suggested to be most likely due to organic acids production [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e]. Similarly, \u003cem\u003eLat. sakei\u003c/em\u003e isolated from \u003cem\u003eSucuk\u003c/em\u003e, a Turkish dry-fermented sausage was reported to exert antifungal activity against \u003cem\u003ePenicillium\u003c/em\u003e and \u003cem\u003eAspergillus\u003c/em\u003e fungi [\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e]. \u003cem\u003eLpb. paraplantarum\u003c/em\u003e antifungal activity found here is compatible to that reported for a non-starter dairy strain [\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e]. Based on the antimicrobial traits of \u003cem\u003eKaddid\u003c/em\u003e isolates, \u003cem\u003eLpb. paraplantarum\u003c/em\u003e K18 and K21, \u003cem\u003eLat. sakei\u003c/em\u003e K25 and \u003cem\u003eLat. sakei\u003c/em\u003e subsp. \u003cem\u003esakei\u003c/em\u003e K22, K26 strains were selected for their promising features mostly due to their ability to inhibit \u003cem\u003eL. monocytogenes\u003c/em\u003e FBUNT. In contrast, \u003cem\u003eE. faecium\u003c/em\u003e strains were not included because of their detrimental traits, virulence factors and antibiotic resistance were not included [\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e, \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eSince bacteriocins from LAB are small peptides (\u0026lt;5 kDa), they are usually thermostable [\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e, \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e]. However, although it was strain dependent, we found a general trend showing a decrease in bacteriocin activity with increasing temperature. BLIS produced by lactobacilli strains from \u003cem\u003eKaddid\u003c/em\u003e, resisted high temperature and pressure during sterilization, indicating they were heat-stable peptides. Similar results were reported for \u003cem\u003eLat. sakei\u003c/em\u003e and \u003cem\u003eLat. curvatus\u003c/em\u003e from fermented meat products and \u003cem\u003eLpb. plantarum\u003c/em\u003e from dairy products, which remained active after 100\u0026deg;C/2 h and 80\u0026deg;C/30 min respectively [\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e, \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e, \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e]. Moreover, BLIS inhibitory activity showed to be higher after exposure to pH 6.5 (400 AU/mL) compared to pH 4.5 (200 AU/mL), Similarly, \u003cem\u003eLpb. plantarum\u003c/em\u003e and \u003cem\u003eLat. sakei\u003c/em\u003e subsp. \u003cem\u003esakei\u003c/em\u003e 2a of dairy and meat origin, showed maximal bacteriocin production at somewhat acidic pH [\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e, \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e]. Confirmation of proteinaceous nature of bacteriocin is essential for the characterization of new antimicrobial peptides. Indeed, inactivation of BLIS inhibitory activity by proteolytic enzymes clearly showed that the antimicrobial substance was of proteinaceous nature; even though the presence of the inhibition halo after enzyme treatment could also suggest other products such as acids or glycoactive compounds as responsible inhibitors. \u003cem\u003eLat. curvatus\u003c/em\u003e CRL705 used as control showed higher inhibitory activity (AU/mL) in agreement with the five protein-encoding genes (lactocin 705, sakacin P, sakacin Q, sakacin X, and sakacin T) responsible for bacteriocin production reported by genome sequencing [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. Although the results obtained from MIC and MBC of BLIS against \u003cem\u003eL. monocytogenes\u003c/em\u003e FBUNT are consistent with previous bacteriocin reported from \u003cem\u003eLactobacillus\u003c/em\u003e [\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e]; antimicrobial compound MICs against the same pathogen species can be variable depending on the strain, the antimicrobial preparation and experimental design used [\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e]. In addition, Class IIa anti-listeria bacteriocins potency would be influenced by the cell-envelope lipid composition of target membranes or differences in the three-dimensional structures of antimicrobial peptides which would determine the varied values of reported MICs [\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe presence of the \u003cem\u003espp\u003c/em\u003eA gene for sakacin P but not \u003cem\u003esap\u003c/em\u003eA for curvacin A detected in \u003cem\u003eLat. sakei\u003c/em\u003e subsp. \u003cem\u003esakei\u003c/em\u003e and \u003cem\u003eLpb. paraplantarum\u003c/em\u003e strains agrees with that described for LAB species isolated from different Argentina meat products [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. Sakacin G is known to possess duplicate structural genes \u003cem\u003eskg\u003c/em\u003eA1 and \u003cem\u003eskg\u003c/em\u003eA2 [\u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e]. The results of sakacin G assessment revealed a PCR fragment of approximately 200 bp, which doubled the anticipated size with the employed primers in this study [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. To validate the identity of this gene, the nucleotide sequence of the PCR fragment amplified from the exemplary producer \u003cem\u003eLat. sakei\u003c/em\u003e subsp. \u003cem\u003esakei\u003c/em\u003e K26, was sequenced. The analysis of the nucleotide sequence showed 100% similarity to the pre-bacteriocin \u003cem\u003eskg\u003c/em\u003eA2 gene (accession number FJ621568). Results from this study correlated with the sakacin P, sakacin G encoding genes reported for \u003cem\u003eLat. sakei\u003c/em\u003e CWBI-B1365 from raw poultry meat, \u003cem\u003eLat. curvatus\u003c/em\u003e ACU-1 from fermented sausages and \u003cem\u003eLat. sakei\u003c/em\u003e R1333 from smoked salmon with high anti-\u003cem\u003eListeria\u003c/em\u003e activity [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e, \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e, \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e53\u003c/span\u003e]. These bacteriocins belong to class IIa which show strong anti-listeria inhibitory effect as well as other food spoilage and pathogenic bacteria; these small peptides have received much attention due to their generally recognized as safe (GRAS) status, their high biological activity, and their heat stability [\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e]. The presence of bacteriocin-related genes does not necessarily indicate the expression of these peptides; horizontal gene transfer mechanisms can explain the variety of genes and the production of multiple bacteriocins by the same culture [\u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eOn the other hand, in meat products the absence of gas, exopolysaccharides (EPS) and biogenic amines (BA) production by starter cultures must be avoided, since they would lead to an indication of spoilage. Although gas production from gluconate was observed for all tested \u003cem\u003eKaddid\u003c/em\u003e lactobacilli, it did not represent a spoilage risk (it was not included among \u003cem\u003eKaddid\u003c/em\u003e preparation additives), whereas gas from glucose was produced by \u003cem\u003eLat. sakei\u003c/em\u003e subsp. \u003cem\u003esakei\u003c/em\u003e K22. The production of ropiness was more notable using glucose than sucrose; \u003cem\u003eLpb. paraplantarum\u003c/em\u003e K18 and K21 were unable to exhibit a ropy phenotype in correlation with their lowest EPS production level. EPS production by LAB showed average yield ranging between 25 to 9800 mg/L [\u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e55\u003c/span\u003e, \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e56\u003c/span\u003e] using different sugar as substrates. Results from this study are in coincidence with the EPS-forming \u003cem\u003eLat. sakei\u003c/em\u003e KS-82 from Turkish \u003cem\u003eSucuk\u003c/em\u003e and the high molecular weight EPS produced by the probiotic \u003cem\u003eLpb. paraplantarum\u003c/em\u003e BGCG11 strains [\u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e57\u003c/span\u003e, \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e]. Although EPS-producing LAB are of technological interest due to their ability to modify food texture and delay color oxidation (Rodr\u0026iacute;guez-S\u0026aacute;nchez et al., 2021), they are considered undesirable for meat products due to slime production. In addition, no BA production was detected among tested lactobacilli, in correlation to that reported for \u003cem\u003eLat. sakei\u003c/em\u003e strains from Italian sausages [\u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e58\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eSafety concerns on the use of LAB strains carrying antibiotic resistance (AR) genes have been raised, as they can transfer them to pathogenic bacteria through horizontal gene transfer mechanisms. Their presence on LAB to be used as autochthonous starter culture might constitute a possible public health hazard, since fermented foods are important vehicles for enormous amounts of living bacteria to enter human body [\u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e59\u003c/span\u003e]. Results revealed high resistance to TET among \u003cem\u003eKaddid\u003c/em\u003e selected lactobacilli. Indeed, TET is among the most widely reported antibiotics for resistant lactobacilli from fermented meats. \u003cem\u003eLat. sakei\u003c/em\u003e and \u003cem\u003eLpb. plantarum\u003c/em\u003e strains from fermented sausages of different origins showed a high incidence of TET resistance and in a less abundance to ERY [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e, \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. However, high resistance to STR was also described for \u003cem\u003eLactobacillaceae\u003c/em\u003e from Mediterranean fermented sausages involving \u003cem\u003eLat. sakei\u003c/em\u003e as dominant species [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. TET resistance of \u003cem\u003eLpb. paraplantarum\u003c/em\u003e K21 disagrees to that reported for a dairy strain [\u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e60\u003c/span\u003e]. The prevalence of \u003cem\u003eKaddid\u003c/em\u003e lactobacilli resistant to TET and STR was supported by the common use of a penicillin/streptomycin complex and/or oxytetracycline for ovine livestock farming in Algeria. These antimicrobials are systematically prescribed to prevent uterine and respiratory infections in postpartum ewes, as well as respiratory and gastrointestinal infections in lambs [\u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e61\u003c/span\u003e]. Widely distributed resistance to aminoglycosides (STR, KAN, GEN) among lactobacilli is likely to be intrinsic as reported by Anisimova \u0026amp; Yarullina [\u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e62\u003c/span\u003e]. This naturally occurring AR is considered relatively safe, since no horizontal transfer between isolates or species was reported [\u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e63\u003c/span\u003e, \u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e64\u003c/span\u003e]. Similarly, an intrinsic resistance of lactobacilli to vancomycin was also reported [\u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e65\u003c/span\u003e]. The high sensitivity to ERY, CLI and CHL observed in lactobacilli from \u003cem\u003eKaddid\u003c/em\u003e agrees with the general susceptibility of lactobacilli to antibiotics inhibiting protein synthesis and resistance to aminoglycosides [\u003cspan citationid=\"CR66\" class=\"CitationRef\"\u003e66\u003c/span\u003e]. In contrast, \u003cem\u003eLat. sakei\u003c/em\u003e strains from Mediterranean sausages were phenotypically resistant to AMP, GEN, KAN, CLI and CHL but not to ERY [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e, \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. On the other hand, the remarkable incidences of AR in enterococci found in this study is in agreement with that reported in animal\u0026rsquo;s feces and raw and fermented meat strains [\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e, \u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e67\u003c/span\u003e, \u003cspan citationid=\"CR68\" class=\"CitationRef\"\u003e68\u003c/span\u003e]. The importance of food enterococci as a reservoir of antibiotic resistance genes and the potential for their genetic transfer to human strains following consumption of uncooked or undercooked contaminated meat is of high concern.\u003c/p\u003e \u003cp\u003eWhen the presence of AR genes was assessed, \u003cem\u003etet\u003c/em\u003eM gene was predominantly found in \u003cem\u003eKaddid\u003c/em\u003e strains. As reported by Fontana et al. [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e], this gene was associated with different MIC values (32\u0026ndash;256 \u0026micro;g/mL) of lactobacilli strains. Indeed, increasing \u003cem\u003etet\u003c/em\u003eM transcript levels were correlated with increased MIC values, suggesting that the expression of this gene is dependent on TET concentration [\u003cspan citationid=\"CR69\" class=\"CitationRef\"\u003e69\u003c/span\u003e]. In this study, a MIC value of 64 \u0026micro;g/mL was found for K21, K22, K25 and K26 lactobacilli strains expressing \u003cem\u003etet\u003c/em\u003eM gene, however \u003cem\u003eLpb. paraplanatarum\u003c/em\u003e K18 had a low MIC (4 \u0026micro;g/mL) and a sensitive phenotype to TET also expressing \u003cem\u003etet\u003c/em\u003eM gene. The correlation between MIC and genetic determinants incidence is strictly strain dependent, however the association between high MICs and the existence of at least one genetic determinant for TET resistant strains was also described [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. The abundance of TET resistance genes in pig feces microbiome and, consequently in the raw meat used for fermented sausages was also reported [\u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e70\u003c/span\u003e, \u003cspan citationid=\"CR71\" class=\"CitationRef\"\u003e71\u003c/span\u003e]. Furthermore, even when \u003cem\u003eLat. sakei\u003c/em\u003e K25 was categorized as phenotypically resistant, none of the \u003cem\u003eKaddid\u003c/em\u003e isolates showed the presence of genes associated with STR resistance suggesting that these genes were not involved in this isolate\u0026rsquo;s resistance. Similarly, \u003cem\u003eLpb. paraplantarum\u003c/em\u003e K18 was phenotypically susceptible to TET and showed to harbor \u003cem\u003etet\u003c/em\u003eM gene. These discrepancies between the resistance phenotype and genotype may be the result of defective expression of resistance genes [\u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e62\u003c/span\u003e]. Altogether, the results from this study confirm the prevalence of \u003cem\u003etet\u003c/em\u003eM gene representing the most widespread resistance determinant in \u003cem\u003eKaddid\u003c/em\u003e strains. Furthermore, it was reported that \u003cem\u003etet\u003c/em\u003eM is located in plasmids suggesting that it has the potential to move between different organisms with an encoding ribosomal protection protein catalyzing the release of tetracycline from the ribosome [73]. Therefore, TET resistance in bacterial strains for food and agricultural applications always constitutes a risk of AR resistance genes spread in the environment.\u003c/p\u003e"},{"header":"5. Conclusions","content":"\u003cp\u003eThe results of this study suggest promising use of autochthonous LAB strains for production of traditional \u003cem\u003eKaddid\u003c/em\u003e. Among evaluated lactobacilli, The strains \u003cem\u003eLpb. paraplantarm\u003c/em\u003e K18 and K21 exhibited desirable \u003cem\u003ein vitro\u003c/em\u003e functional properties namely the inhibitory potential against \u003cem\u003eL. monocytogenes\u003c/em\u003e, low risk as spoilers and free of antibiotic resistance, therefore considered as good candidates to be used as a successful autochthonous functional starter culture to improve \u003cem\u003eKaddid\u003c/em\u003e quality and safety. Further investigation to assess their technological and functional performance is currently undertaken.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eSupplementary Materials:\u0026nbsp;\u003c/strong\u003eTables S1, S2, S3 and S4; Figure S1 and S2.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgments:\u0026nbsp;\u003c/strong\u003eKamel\u0026rsquo;s Boubakri PhD scholarship long stay at CERELA (PNE 2018/2019,\u0026nbsp;N\u0026deg; 256) was supported by the Algerian Ministry of high education and scientific research and the university of\u0026nbsp;M\u0026eacute;d\u0026eacute;a, Algeria. Theauthors thank Dr. Nadia E. Suarez from the sequencing service at CERELA for their help in biomolecular interpretations and Mouna El Hassani PhD at the university of Medea for reviewing the English language of the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eKamel Boubakri:\u0026nbsp;\u003c/strong\u003eConceptualization, Formal analysis, Data curation, Investigation, Methodology, Software, Validation, Visualization, Writing original draft. \u003cstrong\u003eTayeb Idoui:\u0026nbsp;\u003c/strong\u003eConceptualization, Project administration, Validation. Writing review \u0026amp; editing. \u003cstrong\u003eCecilia Aristimu\u0026ntilde;o Ficoseco:\u003c/strong\u003e Formal analysis, Methodology, Resources, Software, Supervision, Validation. \u003cstrong\u003eFranco J. Segli:\u0026nbsp;\u003c/strong\u003eMethodology, Resources. \u003cstrong\u003ePatricia Castellano:\u0026nbsp;\u003c/strong\u003eFunding acquisition, Methodology, Resources.\u003cstrong\u003eLucila Saavedra\u003c/strong\u003e\u003c/a\u003e\u003cstrong\u003e:\u003c/strong\u003e Funding acquisition, Methodology, Resources. Writing review \u0026amp; editing\u003cstrong\u003e. Graciela M. Vignolo:\u0026nbsp;\u003c/strong\u003eConceptualization, Data curation, Funding acquisition, Investigation, Methodology, Project administration, Resources, Supervision, Validation. Writing review \u0026amp; editing. All authors have read and agreed to the published version of the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability:\u003c/strong\u003e The datasets generated during the current study are available in the article and in the online supplementary material.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding:\u003c/strong\u003e No funding was received for conducting this study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of Interest:\u003c/strong\u003e The authors declare no competing interests.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFinancial interests:\u0026nbsp;\u003c/strong\u003eThe authors have no relevant financial or non-financial interests to disclose.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthical Approval Statement:\u0026nbsp;\u003c/strong\u003eThe paper reflects the authors\u0026rsquo; own research and analysis in a truthful and complete manner.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePatient consent statement:\u0026nbsp;\u003c/strong\u003enot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePermission to reproduce material from other sources:\u003c/strong\u003e not applicable\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eBader R, Becila S, Ruiz P, Djeghim F, Sanah I, Boudjellal A, Gatellier P, Portanguen S, Talon R, Leroy S. 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Syst Appl Microbiol. 2003; 26(2): 277\u0026ndash;283. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/org/10.1078/072320203322346137\u003c/span\u003e\u003cspan address=\"10.1078/072320203322346137\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"discover-food","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"discoverfood","sideBox":"Learn more about [Discover Food](https://www.springer.com/44187)","snPcode":"","submissionUrl":"","title":"Discover Food","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Discover Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Kaddid, fermentation, Lpb. paraplantarum, Lat. sakei, bacteriocins, antimicrobial resistance","lastPublishedDoi":"10.21203/rs.3.rs-5005817/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5005817/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eLactic acid bacteria (LAB) from meat dry-salted \u003cem\u003eKaddid\u003c/em\u003e were evaluated using spoilage and safety criteria as a first step of multivariable selection. LAB were isolated, identified and characterized from \u003cem\u003eKaddid\u003c/em\u003e prepared under controlled conditions. After preliminary physiological characterization, isolates (\u003cem\u003en\u003c/em\u003e\u0026thinsp;=\u0026thinsp;39) were subjected to (GTG)5-PCR analysis and 16S rRNA sequencing. Major LAB populations were, \u003cem\u003eLactiplantibacillus paraplantarum\u003c/em\u003e (41%), \u003cem\u003eLatilactobacillus sakei/\u003c/em\u003esubsp. \u003cem\u003esakei\u003c/em\u003e (25%), \u003cem\u003eEnterococcus faecium\u003c/em\u003e (13%), \u003cem\u003ePediococcus acidilactici\u003c/em\u003e (10%), \u003cem\u003eEnterococcus hirae\u003c/em\u003e (6%) and \u003cem\u003eWeissella cibaria\u003c/em\u003e (5%). Antagonistic activity against pathogens/contaminants and yeast/molds showed strains with antilisterial and antifungal activity. Bacteriocin-like inhibitory substances (BLIS) showed high titles (AU/mL) against \u003cem\u003eL. monocytogenes\u003c/em\u003e FBUNT. The examination of bacteriocin genes revealed \u003cem\u003eLpb. paraplantarum\u003c/em\u003e K18, \u003cem\u003eLat. sakei\u003c/em\u003e K25 and \u003cem\u003eLat. sakei\u003c/em\u003e subsp. \u003cem\u003esakei\u003c/em\u003e K22, K26 strains harboring \u003cem\u003eskg\u003c/em\u003eA and \u003cem\u003esak\u003c/em\u003eP structural genes encoding for sakacin G and P bacteriocins. Resistance/susceptibility of lactobacilli strains to antibiotics showed high phenotypic resistance to TET while multi-resistance pattern was displayed by enterococci. Genotypic characterization exhibited only \u003cem\u003etet\u003c/em\u003eM out of 15 assayed genes, conferring resistance to TET. As a quality criterion, no exopolysaccharides and biogenic amines production were detected in the evaluated strains. Based on these results, \u003cem\u003eLpb. paraplantarum\u003c/em\u003e K18 and K21 strains, with inhibitory and antifungal activities, phenotypical susceptibility to the assayed antimicrobial compounds and low risk as spoilers, could be used as functional starter cultures for safe \u003cem\u003eKaddid\u003c/em\u003e production.\u003c/p\u003e","manuscriptTitle":"Autochthonous Lactic Acid Bacteria from Kadidd as Functional Starter Culture to Improve quality and safety","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-10-07 17:09:24","doi":"10.21203/rs.3.rs-5005817/v1","editorialEvents":[{"type":"communityComments","content":1},{"type":"decision","content":"Revision requested","date":"2024-11-14T04:41:31+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-11-13T12:49:54+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"177009412926951390731477251119620235528","date":"2024-11-06T10:36:05+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"222282973788773686840506150602216675103","date":"2024-11-05T10:31:55+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"252506728550341345977786994350013122990","date":"2024-11-05T06:26:37+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-10-20T15:17:02+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"8453969904120054171914410673260262968","date":"2024-10-15T07:04:53+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"240255094058640841605634046406704449042","date":"2024-10-01T13:26:24+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-09-24T06:37:47+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-09-09T11:46:21+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2024-09-05T07:46:09+00:00","index":"","fulltext":""},{"type":"submitted","content":"Discover Food","date":"2024-08-30T19:38:31+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"discover-food","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"discoverfood","sideBox":"Learn more about [Discover Food](https://www.springer.com/44187)","snPcode":"","submissionUrl":"","title":"Discover Food","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Discover Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"6374b140-36e0-4dad-843e-52c0b7003f43","owner":[],"postedDate":"October 7th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2025-08-27T16:34:08+00:00","versionOfRecord":{"articleIdentity":"rs-5005817","link":"https://doi.org/10.1007/s44187-025-00347-0","journal":{"identity":"discover-food","isVorOnly":false,"title":"Discover Food"},"publishedOn":"2025-04-24 00:00:00","publishedOnDateReadable":"April 24th, 2025"},"versionCreatedAt":"2024-10-07 17:09:24","video":"","vorDoi":"10.1007/s44187-025-00347-0","vorDoiUrl":"https://doi.org/10.1007/s44187-025-00347-0","workflowStages":[]},"version":"v1","identity":"rs-5005817","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5005817","identity":"rs-5005817","version":["v1"]},"buildId":"FbvkV6FR0MCFSLy54lSbu","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}