Unveiling the culturable Bacillus sp. from honey in Burkina Faso as a potential source for antimicrobial peptides in plant disease control

Unveiling the culturable Bacillus sp. from honey in Burkina Faso as a potential source for antimicrobial peptides in plant disease control | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Unveiling the culturable Bacillus sp. from honey in Burkina Faso as a potential source for antimicrobial peptides in plant disease control Sonagnon H.S. Kouhounde, Hilarie Bonane, Ulysse Daa Kpode, Soufiane Sanou, and 8 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7280305/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background: Bees and honey are garnering attention as potential sources for isolating beneficial microorganisms. However, the diversity of antibacterial peptides from microorganisms in honey produced in Burkina Faso remains poorly documented. This study aims to explore the diversity of genes detected in Bacillus sp. isolated from honey in Burkina Faso. Results: Here 11 honey samples were used as a matrix to isolate Bacillus spp. bacteria. A total of 40 presumptive Bacillus spp. isolates, preliminarily characterized for Gram staining, catalase tests, and survival under thermal stress, were selected. The RT-PCR with universal 16S primers revealed that all 40 isolates belonged to the genus Bacillus and the presence of genes involved in iturine A, flagellin, TasA, surfactin, subtilin, and mersacidin synthesis. Among the isolates, 65% carry the Bacillus cereus gene. The rest consists (about 40%) of beneficial strains possessing at least one out of six genes and five strains exhibited at least three genes simultaneously. Associations of more than three genes have detected in Bacillus sp. strains, however they are almost absent in Bacillus cereus . Conclusion: Honey can be considered a natural source of beneficial bacteria belonging to Bacillus genus with a broad spectrum of antimicrobial activities useful for plant bioprotection applications. Biological sciences/Biotechnology Biological sciences/Microbiology Biological sciences/Molecular biology Honey Burkina Faso Bacillus sp. genetic diversity RT-PCR antimicrobial peptides Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Background Beekeeping, commonly found in all regions of the globe 1 , is the science and practice of raising bees to exploit their products. It contributes to improving the incomes of populations in the Global South 2 . Beyond honey and its multiple virtues, the maintenance of biodiversity and the pollination of flowering plants 3 are undoubtedly the most valuable ecosystem services provided by bees. Bees, therefore, have both ecological and economic stakes (Varol & Yücel, 2019). In Africa, beekeeping is often a secondary activity but serves as a means of combating poverty through job creation and providing supplementary income (Sawadogo, 2021). Indeed, honey, which is a viscous sweetener 6 produced by bees from nectar or secretion of flowering plants 7 , 8 , is a highly valued commodity nowadays. Its major constituents include 80% of carbohydrates (35% glucose, 40% fructose and 5% sucrose), and water (20%) 9 – 11 , while its minor components include minerals, proteins, vitamins, organic acids, flavonoids, phenolic acids, enzymes and other phytochemicals 12 . The nutritional value of honey depends on its content of mineral substances that are beneficial to humans 13 . Several studies reveal that honey presents many therapeutic traits 14 . Furthermore, honey is an important element in traditional African medicine, where it is used in the treatment of respiratory ailments, memory disorders, fever, burns, gastrointestinal diseases, asthma, infected wounds, and skin ulcers 15 , 16 in both human and animal medicine. Generally, it is considered free of microorganisms. However, it has been reported that yeasts, molds, and aerobic mesophilic bacteria can survive in such a matrix 17 . Studies report that Cladosporium , Aspergillus spp., Penicillium spp., and Bacillus spp. are the main microbial groups found as dominant microorganisms in honey (Terbagou et al., 2024). These microorganisms can originate from nectar, pollen, and other parts of the plant flower 12 . Their presence in honey has been linked to the water content, which is likely to encourage the growth of existing microorganisms. Recent studies conducted in other countries around the world report the presence of a diversity of Bacillus species such as ( Bacillus cereus, Bacillus thuringiensis, Bacillus licheniformis, Bacillus pumilus, Bacillus megaterium and Bacillus coagulans ) on flowers foraged by bees and even in the produced honey 19 – 21 . However, little data exists on the microbial population of honey produced in Burkina Faso, and even less specifically on the Bacillus spp. The objective of the present study was to determine the bacterial population of the genus Bacillus sp. found in honey produced in Burkina Faso. Methods Sampling Sampling was carried out in three provinces (Houet, Tuy, and Kenedougou), chosen to be among the top five honey-producing provinces in Burkina Faso ( Fig. 1 ) . A total of 11 honey samples were collected from producers. In detail, two samples were collected from Houet province, six samples from Kenedougou province, and three samples from Tuy province. The samples were collected in sterile jars and transported in a cooler container to the laboratory at New Dawn University in Bobo. Isolation and purification of presumptive Bacillus sp. Honey samples were treated according to the standard procedure 22 . For each sample, 10 g was taken and diluted into 90 mL of diluent and gently homogenized, then decimal dilutions were obtained. Bacillus sp. were isolated and purified according to Ouoba and co-workers 1 mL of each dilution was plated onto Luria Bertani agar medium (Powder, Liofilchem srl, ITALY) and plates incubated at 37°C for 24 hours 23 . Bacillus cereus ATCC 14579 was used in this study as control strain. The resulting colonies were purified by streaking on the same agar medium several times by successive subculturing on nutrient agar. The isolates were stored at -20°C in nutrient broth supplemented with 20% glycerol. Preliminary characterization of isolates The isolates were described for colony shape, size, color, consistency, and outline, and cultured cells were observed under the microscope (B-292, Optika, Italy) for cell shape, motility, and mode of cell association after Gram staining. Catalase test was performed following the conventional procedure, using a few drops of 3% (v/v) hydrogen peroxide and observing the production of gas bubbles. The sporulation test was carried out according to the method described by Gounina-Allouane and co-workers 24 . In detail, a 24-hour culture was subcultured in nutrient broth and subjected to heat shock at 90°C for 10 minutes in a water bath. The tubes were then placed in an incubator at 37°C for 24 hours. The isolate was declared sporogenous if visual growth was observed. To determine the salt tolerance, the isolates were cultured on LB broth medium (Powder, Liofilchem srl, Italy) supplemented with NaCl at different final concentrations (2.5, 5.0, and 10%), and the results were read by observing bacterial growth after incubation. Detection and quantification of genes following the RT-PCR reaction Fresh bacterial cultures were revived in nutrient broth for 18 hours. Then, 200 µL of chelex (Chelex 100, BIORAD, USA) was dispensed into Eppendorf tubes, and 1 µL of the fresh bacterial culture was added. The cultures were incubated at 56°C for 30–60 min. The mixture was vortexed for 10 sec and returned to incubation at 90°C for 10 min. before being centrifuged at 13,000 rpm for 5 min. Once the extraction was completed, the reaction medium was prepared for the amplification 25 . Eight (8) primer pairs ( Table 1 ) targeting genes (16S RNA, ituA, hag, tasA, srf, sps, mrsH, Bacillus cereus ) described by 26 as beneficial genes for the prevention and control of fungal contamination were used to characterize the 40 selected isolates. Two different amplification programs were used. All PCR amplifications were performed in a 20 µL reaction volume, including 2 µL of genomic DNA and 18 µL of Mix. PCR amplification was performed using a T100 PCR thermocycler (My Cycler, Bio-Rad Laboratories). For amplification of B. cereus gene, PCR was carried out following a 32-cycle program including one (01) cycle of initial DNA denaturation at 95°C for 10 min followed by a set of 30 cycles with denaturation for 40 s at 95°C, hybridization at 59°C for 40 s and elongation for 2 min at 72°C. Then a final elongation (1 cycle) war performed for 6 min at 72°C. Regarding the antifungal genes, the PCR program consists of 32 cycles with an initial denaturation (1 cycle) at 95°C for 3 min, followed by 30 cycles of denaturation for 30 s at 95°C, hybridization at 55°C for 60 s, and elongation for 90 s at 72°C. A final elongation (1 cycle) was performed at 72°C for 5 min. Table 1 Primers used for RT-PCR Sequences (5’ 3’) Genus (pb) Reference AGAGTTTGATCATGGCTCAG ARN 16S 1520 26 , 30 ACGGTTACCTTGTTACGACTT ATGAAAATTTACGGAGTATATATG itu A 743 TTATAACAGCTCTTCATACGTT ATGAGAATCAACCACAATATCGC hag 700 TTAACCTTTAAGCAATTGAAGAAC ATGGGTATGAAAAAGAAATTAAG tas A 719 TTAGTTTTTATCCTCACTGTGA ATGAAGATTTACGGAATTTATATG srf 743 TTATAAAAGCTCTTCGTACGAG ATGTCAAAGTTCGATGATTTCACA sps 707 TTATTTAGAGATTTTGCAGTTACA ATGAGTCAAGAAGCTATCATTCG mrs H 707 TTAACAAATACATTCAGAAGTTAG GCGTTTGTGCTGGTGTAAGT Bacillus cereus 621 CTGCAATCGGAAGGAAGAAG The PCR products were run on a 2% agarose gel at 120 volts for 1 hour and visualized on a transilluminator (TM-20 UV, UVP, France). The 100 bp DNA Ladder ready to load (Solis Biodyne, Estonia) was used as a control size. Antimicrobial resistance The antibiogram based on the determination of minimum inhibitory concentration (MIC) was performed by diffusion using Kirby-Bauer method 27 . The tested antibiotics (Table 2 ) were selected according to the EUCAST (European Committee on Antimicrobial Susceptibility Testing) recommendations for Bacillus species 28 . The inoculum was prepared from an 18-hour culture on nutrient agar. One to two well-isolated and perfectly identical colonies were scraped and transferred into 9 mL of sterile 0.9% saline solution. The entire bacterial suspension was well homogenized and standardized to 0.5 Mac Farland 27 . The bacterial suspension was seeded on the surface of Mueller-Hinton agar (Powder, Liofilchem srl, Italy). Discs pre-impregnated with a known dose of antimicrobial were placed on the surface of the MH agar medium. The distance respected was 1.5 centimeters from a disc to the wall of the Petri dish and 3 cm between two discs. Finally, the dishes were incubated at 37°C for 18 hours aerobically 27 . Table 2 Antibiotics used in the present study Antibiotics Family Code Disc concentration (µg) Critical diameter (mm) S ≥ R˂ Penicillin G Oxacilline Béta-lactamine P Ox 10 1 21 14 22 17 Ciprofloxacine Quinolones CIP 10 21 21 Erythromycine Macrolides E 5 21 18 Vancomycine VA 30 16 16 Gentamicine Amikacine Aminosides CN AK 10 30 18 18 22 22 Trimethoprime-sulfaméthoxazole SXT 25 17 14 After 24 hours of incubation, the reading was performed by measuring the inhibition diameters. The susceptibility profile of each bacterium to the tested antimicrobial compounds was determined using a reading table according to EUCAST, providing the correlation between the inhibition diameter and the MIC, which allowed the bacteria to be classified as susceptible, intermediate, or resistant 28 , 29 . Data Analysis The results were analyzed using R Studio software, after developing an appropriate input mask for the purpose. Multivariate analyses were used to reveal potential differences and associations between the amplified gene fragments. Based on the size of the amplified fragments corresponding to the targeted genes, a hierarchical diagram was generated, allowing the strains to be classified according to the genes they possess. Results Isolation and phenotypic characterization of bacterial isolates A total of 40 isolates belonging to the Bacillus genus were obtained. Identification was based on macroscopic, microscopic, physiological, and molecular characteristics. Macroscopic observation ( Fig. 2 ) of the bacterial colonies, after 24 hours, revealed two colony types: whitish, creamy colonies with regular outlines and large, sticky, rounded colonies. After Gram staining, isolates exhibited Gram-positive bacillus-shaped cells grouped in chains. All isolates were positive for catalase and able to grow after the heat shock. Results on tolerance to NaCl revealed that all isolates were capable of growing in the presence of NaCl up to a concentration of 10%. Amplification of the 16S gene region and genes involved in the synthesis of peptides with antimicrobial activity The identification based on the 16S used primer confirmed that all isolates belong to Bacillus genus. Among them, some possess the ability to synthesize several peptides with antimicrobial activity ( Fig. 3 ) . According to Fig. 3 , it is clear that the isolated strains can be divided into four (04) classes concerning the presence of genes involved in the production of peptides with antimicrobial activity sought. Cluster 1 includes 28 strains. This group, within which B. cereus is strongly represented, is characterized by strains presenting itu A, tas A, Srf , and hag genes (from the most extreme to the least extreme). Cluster 2 included strains positive for the presence of tas A, Srf , and itu A genes. Cluster 3 is composed of strains characterized by high itu A peptide synthesis values. Cluster 4 is composed of strains characterized by the ability to synthesize peptides with antimicrobial activity, such as tas A, Hag , itu A, and Srf . Using the universal primers for Bacillus cereus , 26 of the identified strains (65% of the strains) carried the Bacillus cereus -specific gene ( Fig. 4 ) . The remaining 14 strains were Bacillus sp. strains whose species could not be detected. The search for genes such as itu A (IturinA), hag (Flagellin), tas A (TasA), srf (Surfactin), spa S (Subtilin), and mrs A (Mersacidin) related to the production of antifungal peptides generated the results shown in Table 3 . Overall, each of these genes responsible for the synthesis of bioactive microbial secondary metabolites was identified in our strains. However, a negative PCR result for spa S was obtained in almost all strains. Ten out of the 40 strains (BH1.0, BH1.1, BH1.2, BH2.1, HT3.0, HT3.1, HT3.2, OK9.0, OK10.0, and KK12.2) did not show antifungal genes. Table 3 Distribution of antimicrobial genes in isolated strains Isolates Bacillus cereus Bacillus sp. Detected genes ituA hag tasA srf spS mrsH BH1.0, BH1.1, BH1.2, BH2.1, HT3.0, HT3.1, HT3.2, BT7.0, OK9.0, OK10.0, KK11.2, KK12.2 + - - - - - - - BH2.0 + - + + + - - - BH2.2 + - - - - + - - HT4.0 - + + + - + - - HT4.1 - + + + + + - + HT4.2 - + + + + - - - HT5.0, KK11.3 + - - - + + - - HT5.1 - + + + - - - - HT5.2 - + + + + + - - BT6.0, BT8.2, BT8.3 + - - - + - - - BT6.1 + - - + - - - - BT6.2 - + - - - + - - BT7.1, OK9.1 + - + - - - - - BT7.2 - + + - - - - - BT8.0 + - - + - - - + BT8.1 + - - + + - - - OK9.2, KK12.0 - + - - - - - + OK9.3, KK11.1 - + + + - - - - OK10.1 - + - + + + - - OK10.2, KK12.1 + - - - - - - + OK10.3 - + + + + - - + KK11.0 - + - - - - - - However, gene associations were noted for some strains. These were BH2.0, HT4.1, HT4.2, HT5.2, and OK10.3, which exhibited coproduction of an antifungal substance composed of the macromolecular proteins flagellin, TasA, and the micromolecular lipopeptide (iturinA). Based on the results of this study, we detected the catalase-positive, Gram-positive Bacillus cereus in some of our samples. These different Bacillus cereus strains were tested against eight antibiotics ( Fig. 5 ; Fig. 6 ) selected according to EUCAST recommendations. The Bacillus strains exhibited a variable antimicrobial resistance pattern ( Table 4 ) . Indeed, high resistance (100%) was observed against oxacillin and 84.61% resistance against penicillin. At least 65% of Bacillus strains were resistant to amikacin and trimethoprim-sulfamethoxazole. Low resistance was observed against gentamicin (15.38%), erythromycin (30.76%), vancomycin (19.23%), and ciprofloxacin (1%) against sensitivity rates of 84.61%, 69.23%, 80.76%, and 96.15% respectively. Table 4 Susceptibility of B. cereus strains to tested antibiotics Strains Antibiotics P OX CN VA SXT E AK CIP BH1.0, BT8.1 R R S R R S S S BH1.1 S R S R S R S S BH1.2 R R S S S R S S BH2.0, BH2.1 R R S S S S S S BH2.2 S R S S R S S S HT3.0, OK10.0 R R R S R S R S HT3.1 R R R S S S R S HT3.2, HT5.0, BT8.2 R R S S R S R S BT6.0, OK9.1 R R S S R R R S BT6.1, BT8.0 R R S R R R R S BT7.0, BT7.1 R R S S R R R S BT8.3, OK9.0, KK11.3 R R S S S S R S OK10.2, KK12.2 R R S S R S S S KK11.2 S R S S R S R S KK12.1 S R R S S S S R Legend: (S), Sensitive; (R), Resistant; (P), Penicillin; (CIP), Ciprofloxacin; (OX), Oxacillin; (CN), Gentamicin; (AK), Amikacin; (E), Erythromycin; (VA), Vancomycin; (SXT), Trimethoprim-sulfamethoxazole. Discussion The genus Bacillus sp., a sporulating, Gram-positive bacterium was detected in honey accordingly to previous findings 19 – 21 . Moreover, as already reported, the Bacillus isolated from honey in Asia has been described as able to secrete various peptides and bacteriocins with antifungal activities 26 , 30 . Bacillus spp. devote a large portion of their genome to secondary metabolism, as an adaptive response to environmental competition. Bacillus spores have been detected in soil and the ocean, where complex microbial communities have been described 31 , 32 . By producing secondary metabolites that can inhibit closely related species and other microorganisms in the ecological niche, Bacillus spp. have gained significant survival advantages. In a previous study a variety of secondary metabolites with antibacterial and antifungal properties from Bacillus species, including some non-ribosomal peptides (NRPs) has been found. NRPs are synthesized by non-ribosomal peptide synthetases (NRPSs) and are independent of RNA messenger. NRPs typically undergo extensive modifications, including glycosylation, acylation, and hydroxylation. Due to these modifications, some NRPs are amphiphilic and able to insert into the cell membrane to form pores, such as gramicidin, surfactin, flagellin, iturin, and other lipopeptides. In the present study, 6 pairs carrying genes, such as itu A (IturinA), hag (Flagellin), tas A (TasA), srf (Surfactin), spa S (Subtilin) ​​and mrs A (Mersacidin), which are related to the production of antifungal peptides, were investigated. Although the coproduction of three or more antibacterial molecules is not common, two or three lipopeptides simultaneously produced by the same strain have been described. Our results are consistent with those of previous studies 26 who identified tas A, itu A and hag simultaneously in the same strain of Bacillus sp isolated from honey and demonstrated that these genes conferred properties to control certain fungal species such as Fusarium graminearum , Rhizoctonia solani , and Botrytis cinerea . According to Zhou and co-workers, the first three antifungal peptides have their own characteristics and antifungal spectrum 26 . The iturin family is a cyclic lipopeptide type containing 7 amino acids, with strong antifungal activity. Iturin's antifungal mechanism places its hydrophobic tail into the plasma membrane of indicator cells and automatically assembles to form an ion channel, thus allowing leakage from the cytoplasm. Additionally, iturin A can release electrolytes and polymer aggregates, increase the electrical conductivity and permeability of the biomass membrane, resulting in surface tension effects on the cell membrane, inhibiting pathogen formation 33 . The flagellar filament, formed by a single protein subunit, flagellin, covers 98% of the total length of the flagellum. The flagellin gene is a highly conserved sequence used as a bacterial classification marker. The presence of the gene has been noted in a few strains. According to Nihorimbere and Ongena, lipopeptide production by Bacillus species depends on environmental factors, including temperature, pH, carbon source, and oxygen availability 34 . Antifungal lipopeptides produced by Bacillus spp. may have additional applications. These lipopeptides can modify biofilm formation, motility, and virulence gene expression in various microorganisms 35 , 36 . The TasA protein is one of these lipopeptides. It is involved in sporulation and biofilm formation in Bacillus 37 . These lipopeptides can colonize plant roots, provide plant defenses, and promote plant growth 33 . With the increasing need for biopesticides with high specificity, low environmental persistence, and low toxicity, the industrial exploitation of these biological or naturally derived products as food preservatives and plant protection agents is constantly expanding. Over the past few decades, the incidence of antimicrobial resistance has increased significantly due to the overuse of antibiotics worldwide. According to the results of our study, Bacillus cereus strains isolated from honey exhibited variable antibiotic resistance profiles against the tested molecules. A recent study performed by Noha and co-workers on B. cereus revealed higher susceptibility than our results against gentamicin (93.3%), ciprofloxacin (86.6%), and vancomycin (83.3%) 38 . Here, such resistance found against beta-lactams could be explained by the large prescription of these antibacterials for infection treatments, and therefore for their misuse. Bacteria have therefore been able to develop natural mechanisms to resist beta-lactams 39 , 40 . Generally, Bacillus species secrete beta-lactamase, which hydrolyzes antibiotics 41 , 42 of the beta-lactam class, contributing to the development of resistance. Conclusion The objectives that prompted the present study can be considered achieved. First, the microbiological analysis of the honey revealed the presence of Bacillus . Honey produced in Burkina Faso is a valuable source of interesting bacteria with applications in agronomy and the food industry, where an increasing interest in natural, safe, new, and effective solutions against the harmful effects of global warming has been detected. Results revealed that honey represents a valuable matrix for the isolation of probiotic Bacillus strains for interesting technological applications, being able to synthesize up to five peptides with antifungal effects simultaneously, to be proposed in agriculture to ensure sustainable protection of living beings on the planet. Declarations Funding No funding has been received for this research as financial support. Author Contribution SHSK contributed to the conceptualization and visualization of the study and contributed to the software analysis of the data. HB handled laboratory manipulations, studied, and wrote most of the paper. UDK contributed to collect and software analysis of data. SS contributed to methodology and collect data during the work. VY, RNTM, RCY, YO, WS, KA, MHD and CC reviewed, corrected, and approved the draft article before submission. Acknowledgement The research leading to these results was the result of a collaboration between several entities, including the Applied Biological Sciences Laboratory, the Muraz Center, and the Animal Health and Biotechnology Research and Teaching Laboratory. Data Availability Yes. The datasets generated and analysed during the current study are available in the [PCR-amplified fragments on 2% agarose gel of Bacillus sp. strains using 16S DNA primers] repository, [Data DOI: 10.57760/sciencedb.28857]”. References Abrol, D. P., Shankar, U., Nitin, K. S. & Gowda, G. B. Honeybees and beekeeping: The global scenario. Arthropod Divers. Conserv. Tropics Sub-Tropics . 345–372 10.1007/978-981-10-1518-2_22/TABLES/7 (2016). Jaffé, R. et al. Bees for development: Brazilian survey reveals how to optimize stingless beekeeping. PLoS One 10 , (2015). Hill, R. et al. Biocultural approaches to pollinator conservation. Nat. Sustain. 2 , 214–222 (2019). Varol, E. & Yücel, B. The Effects of Environmental Problems on Honey Bees in View of Sustainable Life . Mellifera vol. 19 (2019). Sawadogo Souhaïbou. Pratiques Apicoles Endogènes et Essai de Co-Développement de Ruches Accessibles Aux Petits Apiculteurs Du Burkina Faso Laboratoire d’Entomologie Fondamentale et Appliquée. (2021). Moriano, M. E. & Alamprese, C. Honey, trehalose and erythritol as sucrose-alternative sweeteners for artisanal ice cream. A pilot study. LWT 75 , 329–334 (2017). Alqarni, A. S. & Honeybee Foraging Nectar Secretion, and Honey Potential of Wild Jujube Trees, Ziziphus nummularia. Neotrop. Entomol. 44 , 232–241 (2015). Adgaba, N. et al. Nectar secretion dynamics and honey production potentials of some major honey plants in Saudi Arabia. Saudi J. Biol. Sci. 24 , 180–191 (2017). Dua, A., Bahman, S., Kelly, S., Dogra, S. & Sharma, K. Authentication of Indian Honey Based on Carbon Stable Isotope Ratio Analysis—Verification of Indian Regulatory Criteria. Foods 14 , 1289 (2025). Faltus, A., Pawłuszkiewicz, K., Mikołajczak, M. & Korgiel, M. Gaweł-Dąbrowska, D. Benefits of Honey Supplementation for Enhancing Health and Athlete Performance. Qual. Sport . 40 , 59354 (2025). Ikegbunam, N. C., Walter, O. J. & Osayi, E. E. Evaluation of the Floral Diversity and Physicochemical Composition of Honey Procured from some Markets in Anambra State, Nigeria. J. Appl. Sci. Environ. Manage. 29 , 961–971 (2025). Cuvillier Miel, Propolis, Gelée Royale: Les Abeilles Alliées de Notre Système Immunitaire. (2015). http://pharmacie.univ-lille2.fr Inaudi, P. et al. Inorganic Composition as Possible Marker for Botanical and Geological Assignment. Molecules vol. 30 Preprint at (2025). https://doi.org/10.3390/molecules30071466 Ogwu, M. C. & Izah, S. C. Honey as a Natural Antimicrobial. Antibiotics 14 , 255 (2025). Abou Auda, M. M. An updated comprehensive review, systematics and biological activity of medicinal plants used in traditional medecines and their common uses in the Gaza Strip, Palestine. Adv. Environ. Biol. 19 , 1–43 (2025). Roy, S., Mishra, B. K., Pattnaik, M. S. & Mahapatra, S. S. Utilization of Apiary Product for Boosting Human Health. (2025). Alippi, A. M., Andez, F., Opez, A. C. L. & L. A. & Diversity of aerobic spore-forming bacteria isolated from fresh bee pollen intended for human consumption in Argentina. J. Apic. Res. 1–8 10.1080/00218839.2021.1960747 (2021). Alippi, A. M., Fernández, L. A. & López, A. C. Diversity of aerobic spore-forming bacteria isolated from fresh bee pollen intended for human consumption in Argentina. J. Apic. Res. 61 , 392–399 (2022). Błónska, D. & Buszewski, B. Characterization of Honey Microbiome Using MALDI-TOF Mass Spectrometry and Physicochemical Study. (2025). 10.3390/molecules30061266 Cirtwill, A. R. & Wirta, H. DNA in honey could describe the changes in flower visits and microbe encounters of honey bees over decades. Sci. Rep. 10.1038/s41598-025-93315-8 (2025). Wang, M., Zhao, W., Zhou, D. & Huang, J. Bacterial diversity in the honey sac during bee foraging on winter-flowering flora and dominant Bacillus subtilis inhibits Hafnia alvei. Front. Insect Sci. 5 , 1555434 (2025). ISO 6887-1. International Organization for Standardization. No.6887-1. Microbiology of the Food Chain — Preparation of Test Samples, Initial Suspension and Decimal Dilutions for Microbiological Examination — Part 1: General Rules for the Preparation of the Initial Suspension and Decimal Dilutions.. (2017). Ouoba, L. I. I., Parkouda, C., Diawara, B., Scotti, C. & Varnam, A. H. Identification of Bacillus spp. from Bikalga, fermented seeds of Hibiscus sabdariffa: Phenotypic and genotypic characterization. J. Appl. Microbiol. 104 , 122–131 (2008). Gounina-Allouane, R., Broussolle, V. & Carlin, F. Influence of the sporulation temperature on the impact of the nutrients inosine and L-alanine on Bacillus cereus spore germination. Food Microbiol. 25 , 202–206 (2008). Susilowati, R., Sabdono, A. & Widowati, I. Isolation and Characterization of Bacteria Associated with Brown Algae Sargassum spp. from Panjang Island and their Antibacterial Activities. Procedia Environ. Sci. 23 , 240–246 (2015). Zhou, Z. et al. Investigation of antibacterial activity of Bacillus spp. isolated from the feces of Giant Panda and characterization of their antimicrobial gene distributions. World J. Microbiol. Biotechnol. 30 , 3129–3136 (2014). CASFM. Comité de l ’ antibiogramme de la Société Française de Microbiologie Recommandations 2022. (2022). EUCAST. Bacillus spp. Calibration of zone diameter breakpoints to MIC values. (2023). https://www.eucast.org/fileadmin/src/media/PDFs/EUCAST_files/Disk_criteria/Validation_2023/Bacillus_v_2.1_February_2023. pdf 1–17. Mills, E., Sullivan, E. & Kovac, J. Comparative Analysis of Bacillus cereus Group Isolates’ Resistance Using Disk Diffusion and Broth Microdilution and the Correlation between Antimicrobial Resistance Phenotypes and Genotypes. (2022). Zhao, X. et al. Isolation and identification of antifungal peptides from bacillus BH072, a novel bacterium isolated from honey. Microbiol. Res. 168 , 598–606 (2013). Chen, M., Takano, C., Nakashima, K., Gowthaman, S. & Kawasaki, S. Exploration of ureolytic airborne bacteria for biocementation applications from different climate zones in Japan. Sci. Rep. 15 , 7536 (2025). Al-Abdulsalam, H., Almalki, M. & Khalifa, A. Isolation and characterization of antimicrobial metabolites producing bacteria from soils in Al-Ahsa, Saudi Arabia. Journal Microbiol. Biotechnol. Food Sciences 14 , (2025). Markelova, N. & Chumak, A. Antimicrobial Activity of Bacillus Cyclic Lipopeptides and Their Role in the Host Adaptive Response to Changes in Environmental Conditions. Int. J. Mol. Sci. 26 , 336 (2025). Nihorimbere, V. & Ongena, M. Isolation of Plant growth-promoting Bacillus Strains with Biocontrol Activity In vitro. Merit Res. J. Microbiol. Biol. Sci. 5 , 2408–7076 (2017). Kumar, M. et al. Unraveling the Bioactivities and Immunomodulatory Potential of Postbiotics Derived from Bacillus subtilis and B. amyloliquefaciens for Aquaculture. Probiotics Antimicrob. Proteins 1–15 (2025). Roque-Borda, C. A. et al. Antimicrobial Peptides: A Promising Alternative to Conventional Antimicrobials for Combating Polymicrobial Biofilms. Adv. Sci. 12 , 2410893 (2025). Kim et al. Production of Biosurfactant Lipopeptides Iturin A, Fengycin, and Surfactin A from Bacillus subtilis CMB32 for Control of Colletotrichum gloeosporioides.pdf. Preprint at (2010). Noha et al. Identification, characterization and antibiotic susceptibility testing for Bacillus species. 14 , 484–494 (2022). Elshobary, M. E. et al. Combating Antibiotic Resistance: Mechanisms, Multidrug-Resistant Pathogens, and Novel Therapeutic Approaches: An Updated Review. Pharmaceuticals 18 , 402 (2025). Vivekanandan, K. E., Kumar, P. V., Jaysree, R. C. & Rajeshwari, T. Exploring Molecular Mechanisms of Drug Resistance in Bacteria and Progressions in CRISPR/Cas9-Based Genome Expurgation Solutions. Glob Med. Genet 100042 (2025). Alemayehu, T. et al. Phenotypic identification of Metallo-ß-lactamase resistance Gram negative bacteria from a clinical specimen in Sidama, Ethiopia. PLoS One . 20 , e0313431 (2025). Geremia, N. et al. Rare or Unusual Non-Fermenting Gram-Negative Bacteria: Therapeutic Approach and Antibiotic Treatment Options. Antibiotics 14 , 306 (2025). Additional Declarations No competing interests reported. Supplementary Files Supportinginformation2.docx Cite Share Download PDF Status: Posted Version 1 posted 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. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-7280305","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":500864793,"identity":"e0cc2787-7083-42c7-b1d7-37d46725814f","order_by":0,"name":"Sonagnon H.S. Kouhounde","email":"data:image/png;base64,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","orcid":"","institution":"Université Aube Nouvelle","correspondingAuthor":true,"prefix":"","firstName":"Sonagnon","middleName":"H.S.","lastName":"Kouhounde","suffix":""},{"id":500864794,"identity":"d0d516ed-50a3-4892-8c5d-12a65e73e57b","order_by":1,"name":"Hilarie Bonane","email":"","orcid":"","institution":"Université Aube Nouvelle","correspondingAuthor":false,"prefix":"","firstName":"Hilarie","middleName":"","lastName":"Bonane","suffix":""},{"id":500864795,"identity":"6c839ab0-abc4-4434-bf68-db978e122e17","order_by":2,"name":"Ulysse Daa Kpode","email":"","orcid":"","institution":"University of Abomey-Calavi","correspondingAuthor":false,"prefix":"","firstName":"Ulysse","middleName":"Daa","lastName":"Kpode","suffix":""},{"id":500864796,"identity":"56176a1c-724f-4d39-ba9d-4ea2e5c993de","order_by":3,"name":"Soufiane Sanou","email":"","orcid":"","institution":"Centre Muraz","correspondingAuthor":false,"prefix":"","firstName":"Soufiane","middleName":"","lastName":"Sanou","suffix":""},{"id":500864797,"identity":"a30faedd-3d5d-49e7-95b0-cbd3a933103c","order_by":4,"name":"Victorien Yameogo","email":"","orcid":"","institution":"Nazi Boni University","correspondingAuthor":false,"prefix":"","firstName":"Victorien","middleName":"","lastName":"Yameogo","suffix":""},{"id":500864798,"identity":"086f32a6-695d-4ff8-8490-d0790321e8ae","order_by":5,"name":"Roland Nag-Tiero Meda","email":"","orcid":"","institution":"Nazi Boni University","correspondingAuthor":false,"prefix":"","firstName":"Roland","middleName":"Nag-Tiero","lastName":"Meda","suffix":""},{"id":500864799,"identity":"bc79eb98-072c-4cc5-b62e-04b9121737f0","order_by":6,"name":"Rene Comlan Yaovi","email":"","orcid":"","institution":"Nazi Boni University","correspondingAuthor":false,"prefix":"","firstName":"Rene","middleName":"Comlan","lastName":"Yaovi","suffix":""},{"id":500864800,"identity":"0835c5b0-a3ee-4fd9-b4fa-f906df59acd0","order_by":7,"name":"Yaser Ouattara","email":"","orcid":"","institution":"Université Aube Nouvelle","correspondingAuthor":false,"prefix":"","firstName":"Yaser","middleName":"","lastName":"Ouattara","suffix":""},{"id":500864801,"identity":"b72f8967-78e7-4ade-9bc0-f9fff1ecd438","order_by":8,"name":"William A. Sib","email":"","orcid":"","institution":"Université Aube Nouvelle","correspondingAuthor":false,"prefix":"","firstName":"William","middleName":"A.","lastName":"Sib","suffix":""},{"id":500864802,"identity":"ef9e7152-ab41-476d-a2b8-dfac15a6ed97","order_by":9,"name":"Kifouli Adeoti","email":"","orcid":"","institution":"University of Abomey-Calavi","correspondingAuthor":false,"prefix":"","firstName":"Kifouli","middleName":"","lastName":"Adeoti","suffix":""},{"id":500864803,"identity":"96a992ab-26f9-4bc3-9436-bef921afdf5f","order_by":10,"name":"Mamoudou H. Dicko","email":"","orcid":"","institution":"Université Joseph Ki-Zerbo","correspondingAuthor":false,"prefix":"","firstName":"Mamoudou","middleName":"H.","lastName":"Dicko","suffix":""},{"id":500864804,"identity":"c2aa4c0a-c78c-42bc-a2f8-7a29fa2c1fa4","order_by":11,"name":"Cinzia Caggia","email":"","orcid":"","institution":"University of Catania","correspondingAuthor":false,"prefix":"","firstName":"Cinzia","middleName":"","lastName":"Caggia","suffix":""}],"badges":[],"createdAt":"2025-08-02 20:53:11","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7280305/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7280305/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":89634578,"identity":"95f2c8d4-b011-4091-befc-ce17cabe0ed4","added_by":"auto","created_at":"2025-08-22 07:07:38","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":112147,"visible":true,"origin":"","legend":"\u003cp\u003eSample collection sites\u003c/p\u003e","description":"","filename":"1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7280305/v1/455d4b5b0c2fb9d247d1c98c.jpg"},{"id":89634590,"identity":"9403a4dd-a88e-46a7-8ea2-65968ce33e31","added_by":"auto","created_at":"2025-08-22 07:07:38","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":524391,"visible":true,"origin":"","legend":"\u003cp\u003eMacroscopic appearance of Bacillus colonies. Panel \u003cstrong\u003eA: \u003c/strong\u003eSlimy colonies; Panel \u003cstrong\u003eB:\u003c/strong\u003e Creamy colonies\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-7280305/v1/a4cdba45d666a9a42312ff74.png"},{"id":89635785,"identity":"09582307-cb67-46ef-ac35-ae255766c2fd","added_by":"auto","created_at":"2025-08-22 07:15:38","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":32849,"visible":true,"origin":"","legend":"\u003cp\u003eDendrogram showing a hierarchical classification of strains into four clusters based on the size of amplified genes and their potential to synthesize one or more antifungal peptides. Cluster 1 groups the KK12.0, OK9.2, KK11.0, KK12.1, OK10.2, BT8.0, BT6.1, BH1.1, BH1.0, HT3.2, HT3.1, BT7.0, OK9.0, OK10.0, KK11.2, KK12.2, HT3.0, BH2.1, BH1.2 strains; Cluster 2 groups BT6.2, BH2.2, KK11.3, HT5.0, BT8.2, BT6.0, BT8.3, BT8.1 strains; Cluster 3 groups OK9.3, HT5.1, KK11.1, HT4.0, OK9.1, BT7.1, BT7.0 strains, and Cluster 4 groups HT4.1, HT5.2, OK10.1, HT4.2, BH2.0, OK10.3 strains.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-7280305/v1/95ad77f8465480896cfedace.png"},{"id":89635787,"identity":"54a4b21f-788a-41a1-9db4-4b63d597314a","added_by":"auto","created_at":"2025-08-22 07:15:38","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":450628,"visible":true,"origin":"","legend":"\u003cp\u003ePCR-amplified fragments on 2% agarose gel of \u003cem\u003eB. cereus\u003c/em\u003e isolated from honey\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-7280305/v1/59e04e57aca1221573b8f01d.png"},{"id":89634584,"identity":"63cec3cb-a4a9-4507-92cd-141f7578edab","added_by":"auto","created_at":"2025-08-22 07:07:38","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":276778,"visible":true,"origin":"","legend":"\u003cp\u003eAntibiogram of B. cereus against the tested antibiotics\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-7280305/v1/e41ea59e1104fd29afa0709f.png"},{"id":89634579,"identity":"698015af-391d-4061-a74d-76ecbf4cc6fb","added_by":"auto","created_at":"2025-08-22 07:07:38","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":99198,"visible":true,"origin":"","legend":"\u003cp\u003eSusceptibility of tested antibiotics with isolated Bacillus cereus strains\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-7280305/v1/bc5af256ad31482ce849ba89.png"},{"id":103720845,"identity":"26ba6fe2-a90e-4d8e-955e-074f7229e8a7","added_by":"auto","created_at":"2026-03-02 07:11:58","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2856775,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7280305/v1/9eaa2e2e-212c-4995-ac89-fd0ad4f27138.pdf"},{"id":89636677,"identity":"d2ea2976-3cbd-46fa-b33e-9ccd3e69bfca","added_by":"auto","created_at":"2025-08-22 07:23:38","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":389570,"visible":true,"origin":"","legend":"","description":"","filename":"Supportinginformation2.docx","url":"https://assets-eu.researchsquare.com/files/rs-7280305/v1/42f66cd359300f5aa8515366.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"\u003cp\u003eUnveiling the culturable \u003cem\u003eBacillus \u003c/em\u003esp. from honey in Burkina Faso as a potential source for antimicrobial peptides in plant disease control\u003c/p\u003e","fulltext":[{"header":"Background","content":"\u003cp\u003eBeekeeping, commonly found in all regions of the globe \u003csup\u003e\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e, is the science and practice of raising bees to exploit their products. It contributes to improving the incomes of populations in the Global South \u003csup\u003e\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e\u003c/sup\u003e. Beyond honey and its multiple virtues, the maintenance of biodiversity and the pollination of flowering plants \u003csup\u003e\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e\u003c/sup\u003e are undoubtedly the most valuable ecosystem services provided by bees. Bees, therefore, have both ecological and economic stakes (Varol \u0026amp; Yücel, 2019). In Africa, beekeeping is often a secondary activity but serves as a means of combating poverty through job creation and providing supplementary income (Sawadogo, 2021).\u003c/p\u003e\u003cp\u003eIndeed, honey, which is a viscous sweetener \u003csup\u003e\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u003c/sup\u003e produced by bees from nectar or secretion of flowering plants \u003csup\u003e\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e,\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e\u003c/sup\u003e, is a highly valued commodity nowadays. Its major constituents include 80% of carbohydrates (35% glucose, 40% fructose and 5% sucrose), and water (20%) \u003csup\u003e\u003cspan additionalcitationids=\"CR10\" citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e–\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u003c/sup\u003e, while its minor components include minerals, proteins, vitamins, organic acids, flavonoids, phenolic acids, enzymes and other phytochemicals \u003csup\u003e\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003eThe nutritional value of honey depends on its content of mineral substances that are beneficial to humans \u003csup\u003e\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e\u003c/sup\u003e. Several studies reveal that honey presents many therapeutic traits \u003csup\u003e\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e\u003c/sup\u003e. Furthermore, honey is an important element in traditional African medicine, where it is used in the treatment of respiratory ailments, memory disorders, fever, burns, gastrointestinal diseases, asthma, infected wounds, and skin ulcers \u003csup\u003e\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e,\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e\u003c/sup\u003e in both human and animal medicine.\u003c/p\u003e\u003cp\u003eGenerally, it is considered free of microorganisms. However, it has been reported that yeasts, molds, and aerobic mesophilic bacteria can survive in such a matrix \u003csup\u003e\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e\u003c/sup\u003e. Studies report that \u003cem\u003eCladosporium\u003c/em\u003e, \u003cem\u003eAspergillus\u003c/em\u003e spp., \u003cem\u003ePenicillium\u003c/em\u003e spp., and \u003cem\u003eBacillus\u003c/em\u003e spp. are the main microbial groups found as dominant microorganisms in honey (Terbagou et al., 2024). These microorganisms can originate from nectar, pollen, and other parts of the plant flower \u003csup\u003e\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e\u003c/sup\u003e. Their presence in honey has been linked to the water content, which is likely to encourage the growth of existing microorganisms.\u003c/p\u003e\u003cp\u003eRecent studies conducted in other countries around the world report the presence of a diversity of \u003cem\u003eBacillus\u003c/em\u003e species such as (\u003cem\u003eBacillus cereus, Bacillus thuringiensis, Bacillus licheniformis, Bacillus pumilus, Bacillus megaterium\u003c/em\u003e and \u003cem\u003eBacillus coagulans\u003c/em\u003e) on flowers foraged by bees and even in the produced honey \u003csup\u003e\u003cspan additionalcitationids=\"CR20\" citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e–\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e\u003c/sup\u003e. However, little data exists on the microbial population of honey produced in Burkina Faso, and even less specifically on the \u003cem\u003eBacillus\u003c/em\u003e spp.\u003c/p\u003e\u003cp\u003eThe objective of the present study was to determine the bacterial population of the genus \u003cem\u003eBacillus\u003c/em\u003e sp. found in honey produced in Burkina Faso.\u003c/p\u003e"},{"header":"Methods","content":"\u003cp\u003e\u003cb\u003eSampling\u003c/b\u003e\u003c/p\u003e\u003cp\u003eSampling was carried out in three provinces (Houet, Tuy, and Kenedougou), chosen to be among the top five honey-producing provinces in Burkina Faso \u003cb\u003e(\u003c/b\u003eFig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e\u003cb\u003e)\u003c/b\u003e. A total of 11 honey samples were collected from producers. In detail, two samples were collected from Houet province, six samples from Kenedougou province, and three samples from Tuy province. The samples were collected in sterile jars and transported in a cooler container to the laboratory at New Dawn University in Bobo.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003eIsolation and purification of presumptive\u003c/b\u003e \u003cb\u003eBacillus\u003c/b\u003e \u003cb\u003esp.\u003c/b\u003e\u003c/p\u003e\u003cp\u003eHoney samples were treated according to the standard procedure \u003csup\u003e\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e\u003c/sup\u003e. For each sample, 10 g was taken and diluted into 90 mL of diluent and gently homogenized, then decimal dilutions were obtained.\u003c/p\u003e\u003cp\u003e\u003cem\u003eBacillus\u003c/em\u003e sp. were isolated and purified according to Ouoba and co-workers 1 mL of each dilution was plated onto Luria Bertani agar medium (Powder, Liofilchem srl, ITALY) and plates incubated at 37°C for 24 hours \u003csup\u003e\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e\u003c/sup\u003e. \u003cem\u003eBacillus cereus\u003c/em\u003e ATCC 14579 was used in this study as control strain.\u003c/p\u003e\u003cp\u003eThe resulting colonies were purified by streaking on the same agar medium several times by successive subculturing on nutrient agar. The isolates were stored at -20°C in nutrient broth supplemented with 20% glycerol.\u003c/p\u003e\u003cp\u003e\u003cb\u003ePreliminary characterization of isolates\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe isolates were described for colony shape, size, color, consistency, and outline, and cultured cells were observed under the microscope (B-292, Optika, Italy) for cell shape, motility, and mode of cell association after Gram staining. Catalase test was performed following the conventional procedure, using a few drops of 3% (v/v) hydrogen peroxide and observing the production of gas bubbles. The sporulation test was carried out according to the method described by Gounina-Allouane and co-workers \u003csup\u003e\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e\u003c/sup\u003e. In detail, a 24-hour culture was subcultured in nutrient broth and subjected to heat shock at 90°C for 10 minutes in a water bath. The tubes were then placed in an incubator at 37°C for 24 hours. The isolate was declared sporogenous if visual growth was observed.\u003c/p\u003e\u003cp\u003eTo determine the salt tolerance, the isolates were cultured on LB broth medium (Powder, Liofilchem srl, Italy) supplemented with NaCl at different final concentrations (2.5, 5.0, and 10%), and the results were read by observing bacterial growth after incubation.\u003c/p\u003e\u003cp\u003e\u003cb\u003eDetection and quantification of genes following the RT-PCR reaction\u003c/b\u003e\u003c/p\u003e\u003cp\u003eFresh bacterial cultures were revived in nutrient broth for 18 hours. Then, 200 µL of chelex (Chelex 100, BIORAD, USA) was dispensed into Eppendorf tubes, and 1 µL of the fresh bacterial culture was added. The cultures were incubated at 56°C for 30–60 min. The mixture was vortexed for 10 sec and returned to incubation at 90°C for 10 min. before being centrifuged at 13,000 rpm for 5 min. Once the extraction was completed, the reaction medium was prepared for the amplification \u003csup\u003e\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003eEight (8) primer pairs \u003cb\u003e(\u003c/b\u003eTable\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e\u003cb\u003e)\u003c/b\u003e targeting genes (16S RNA, \u003cem\u003eituA, hag, tasA, srf, sps, mrsH, Bacillus cereus\u003c/em\u003e) described by\u003csup\u003e\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e\u003c/sup\u003e as beneficial genes for the prevention and control of fungal contamination were used to characterize the 40 selected isolates. Two different amplification programs were used. All PCR amplifications were performed in a 20 µL reaction volume, including 2 µL of genomic DNA and 18 µL of Mix. PCR amplification was performed using a T100 PCR thermocycler (My Cycler, Bio-Rad Laboratories). For amplification of \u003cem\u003eB. cereus\u003c/em\u003e gene, PCR was carried out following a 32-cycle program including one (01) cycle of initial DNA denaturation at 95°C for 10 min followed by a set of 30 cycles with denaturation for 40 s at 95°C, hybridization at 59°C for 40 s and elongation for 2 min at 72°C. Then a final elongation (1 cycle) war performed for 6 min at 72°C. Regarding the antifungal genes, the PCR program consists of 32 cycles with an initial denaturation (1 cycle) at 95°C for 3 min, followed by 30 cycles of denaturation for 30 s at 95°C, hybridization at 55°C for 60 s, and elongation for 90 s at 72°C. A final elongation (1 cycle) was performed at 72°C for 5 min.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cdiv class=\"gridtable\"\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=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\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\u003ePrimers used for RT-PCR\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"4\"\u003e\u003c/colgroup\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSequences (5’ 3’)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGenus\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003e(pb)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eReference\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eAGAGTTTGATCATGGCTCAG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003e\u003cb\u003eARN 16S\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003e\u003cb\u003e1520\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\" morerows=\"15\" rowspan=\"16\"\u003e\u003cp\u003e\u003csup\u003e\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e,\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eACGGTTACCTTGTTACGACTT\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eATGAAAATTTACGGAGTATATATG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003e\u003cb\u003eitu\u003c/b\u003e\u003cb\u003eA\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003e\u003cb\u003e743\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTTATAACAGCTCTTCATACGTT\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eATGAGAATCAACCACAATATCGC\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003e\u003cb\u003ehag\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003e\u003cb\u003e700\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTTAACCTTTAAGCAATTGAAGAAC\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eATGGGTATGAAAAAGAAATTAAG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003e\u003cb\u003etas\u003c/b\u003e\u003cb\u003eA\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003e\u003cb\u003e719\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTTAGTTTTTATCCTCACTGTGA\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eATGAAGATTTACGGAATTTATATG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003e\u003cb\u003esrf\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003e\u003cb\u003e743\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTTATAAAAGCTCTTCGTACGAG\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eATGTCAAAGTTCGATGATTTCACA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003e\u003cb\u003esps\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003e\u003cb\u003e707\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTTATTTAGAGATTTTGCAGTTACA\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eATGAGTCAAGAAGCTATCATTCG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003e\u003cb\u003emrs\u003c/b\u003e\u003cb\u003eH\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003e\u003cb\u003e707\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTTAACAAATACATTCAGAAGTTAG\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGCGTTTGTGCTGGTGTAAGT\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003e\u003cb\u003eBacillus cereus\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003e\u003cb\u003e621\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eCTGCAATCGGAAGGAAGAAG\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/table\u003e\u003c/div\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eThe PCR products were run on a 2% agarose gel at 120 volts for 1 hour and visualized on a transilluminator (TM-20 UV, UVP, France). The 100 bp DNA Ladder ready to load (Solis Biodyne, Estonia) was used as a control size.\u003c/p\u003e\u003cp\u003e\u003cb\u003eAntimicrobial resistance\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe antibiogram based on the determination of minimum inhibitory concentration (MIC) was performed by diffusion using Kirby-Bauer method \u003csup\u003e\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e\u003c/sup\u003e. The tested antibiotics (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e) were selected according to the EUCAST (European Committee on Antimicrobial Susceptibility Testing) recommendations for \u003cem\u003eBacillus\u003c/em\u003e species \u003csup\u003e\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e\u003c/sup\u003e. The inoculum was prepared from an 18-hour culture on nutrient agar. One to two well-isolated and perfectly identical colonies were scraped and transferred into 9 mL of sterile 0.9% saline solution. The entire bacterial suspension was well homogenized and standardized to 0.5 Mac Farland \u003csup\u003e\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e\u003c/sup\u003e. The bacterial suspension was seeded on the surface of Mueller-Hinton agar (Powder, Liofilchem srl, Italy). Discs pre-impregnated with a known dose of antimicrobial were placed on the surface of the MH agar medium. The distance respected was 1.5 centimeters from a disc to the wall of the Petri dish and 3 cm between two discs. Finally, the dishes were incubated at 37°C for 18 hours aerobically \u003csup\u003e\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cdiv class=\"gridtable\"\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=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\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\u003eAntibiotics used in the present study\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"5\"\u003e\u003c/colgroup\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eAntibiotics\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eFamily\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eCode\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eDisc concentration (µg)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eCritical diameter (mm)\u003c/p\u003e\u003cp\u003eS ≥ R˂\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePenicillin G\u003c/p\u003e\u003cp\u003eOxacilline\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eBéta-lactamine\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eP\u003c/p\u003e\u003cp\u003eOx\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e10\u003c/p\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e21 14\u003c/p\u003e\u003cp\u003e22 17\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eCiprofloxacine\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eQuinolones\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eCIP\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e21 21\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eErythromycine\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eMacrolides\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eE\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e21 18\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eVancomycine\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eVA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e30\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e16 16\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGentamicine\u003c/p\u003e\u003cp\u003eAmikacine\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eAminosides\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eCN\u003c/p\u003e\u003cp\u003eAK\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e10\u003c/p\u003e\u003cp\u003e30\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e18 18\u003c/p\u003e\u003cp\u003e22 22\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTrimethoprime-sulfaméthoxazole\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eSXT\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e25\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e17 14\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/table\u003e\u003c/div\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eAfter 24 hours of incubation, the reading was performed by measuring the inhibition diameters. The susceptibility profile of each bacterium to the tested antimicrobial compounds was determined using a reading table according to EUCAST, providing the correlation between the inhibition diameter and the MIC, which allowed the bacteria to be classified as susceptible, intermediate, or resistant \u003csup\u003e\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e,\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\u003ch2\u003eData Analysis\u003c/h2\u003e\u003cp\u003eThe results were analyzed using R Studio software, after developing an appropriate input mask for the purpose. Multivariate analyses were used to reveal potential differences and associations between the amplified gene fragments. Based on the size of the amplified fragments corresponding to the targeted genes, a hierarchical diagram was generated, allowing the strains to be classified according to the genes they possess.\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003e\u003cb\u003eIsolation and phenotypic characterization of bacterial isolates\u003c/b\u003e\u003c/p\u003e\u003cp\u003eA total of 40 isolates belonging to the \u003cem\u003eBacillus\u003c/em\u003e genus were obtained. Identification was based on macroscopic, microscopic, physiological, and molecular characteristics. Macroscopic observation \u003cb\u003e(\u003c/b\u003eFig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e\u003cb\u003e)\u003c/b\u003e of the bacterial colonies, after 24 hours, revealed two colony types: whitish, creamy colonies with regular outlines and large, sticky, rounded colonies. After Gram staining, isolates exhibited Gram-positive bacillus-shaped cells grouped in chains. All isolates were positive for catalase and able to grow after the heat shock. Results on tolerance to NaCl revealed that all isolates were capable of growing in the presence of NaCl up to a concentration of 10%.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003eAmplification of the 16S gene region and genes involved in the synthesis of peptides with antimicrobial activity\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe identification based on the 16S used primer confirmed that all isolates belong to \u003cem\u003eBacillus\u003c/em\u003e genus. Among them, some possess the ability to synthesize several peptides with antimicrobial activity \u003cb\u003e(\u003c/b\u003eFig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e\u003cb\u003e)\u003c/b\u003e. According to Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e, it is clear that the isolated strains can be divided into four (04) classes concerning the presence of genes involved in the production of peptides with antimicrobial activity sought. Cluster 1 includes 28 strains. This group, within which \u003cem\u003eB. cereus\u003c/em\u003e is strongly represented, is characterized by strains presenting \u003cem\u003eitu\u003c/em\u003eA, \u003cem\u003etas\u003c/em\u003eA, \u003cem\u003eSrf\u003c/em\u003e, and \u003cem\u003ehag\u003c/em\u003e genes (from the most extreme to the least extreme). Cluster 2 included strains positive for the presence of \u003cem\u003etas\u003c/em\u003eA, \u003cem\u003eSrf\u003c/em\u003e, and \u003cem\u003eitu\u003c/em\u003eA genes. Cluster 3 is composed of strains characterized by high \u003cem\u003eitu\u003c/em\u003eA peptide synthesis values. Cluster 4 is composed of strains characterized by the ability to synthesize peptides with antimicrobial activity, such as \u003cem\u003etas\u003c/em\u003eA, \u003cem\u003eHag\u003c/em\u003e, \u003cem\u003eitu\u003c/em\u003eA, and \u003cem\u003eSrf\u003c/em\u003e.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eUsing the universal primers for \u003cem\u003eBacillus cereus\u003c/em\u003e, 26 of the identified strains (65% of the strains) carried the \u003cem\u003eBacillus cereus\u003c/em\u003e-specific gene \u003cb\u003e(\u003c/b\u003eFig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e\u003cb\u003e)\u003c/b\u003e. The remaining 14 strains were \u003cem\u003eBacillus\u003c/em\u003e sp. strains whose species could not be detected.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eThe search for genes such as \u003cem\u003eitu\u003c/em\u003eA (IturinA), \u003cem\u003ehag\u003c/em\u003e (Flagellin), \u003cem\u003etas\u003c/em\u003eA (TasA), \u003cem\u003esrf\u003c/em\u003e (Surfactin), \u003cem\u003espa\u003c/em\u003eS (Subtilin), and \u003cem\u003emrs\u003c/em\u003eA (Mersacidin) related to the production of antifungal peptides generated the results shown in Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e. Overall, each of these genes responsible for the synthesis of bioactive microbial secondary metabolites was identified in our strains. However, a negative PCR result for \u003cem\u003espa\u003c/em\u003eS was obtained in almost all strains. Ten out of the 40 strains (BH1.0, BH1.1, BH1.2, BH2.1, HT3.0, HT3.1, HT3.2, OK9.0, OK10.0, and KK12.2) did not show antifungal genes.\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\u003eDistribution of antimicrobial genes in isolated strains\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"9\"\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\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eIsolates\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003e\u003cem\u003eBacillus cereus\u003c/em\u003e\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003e\u003cem\u003eBacillus\u003c/em\u003e sp.\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colspan=\"6\" nameend=\"c9\" namest=\"c4\"\u003e\u003cp\u003eDetected genes\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cem\u003eituA\u003c/em\u003e\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u003cem\u003ehag\u003c/em\u003e\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003e\u003cem\u003etasA\u003c/em\u003e\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003e\u003cem\u003esrf\u003c/em\u003e\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c8\"\u003e\u003cp\u003e\u003cem\u003espS\u003c/em\u003e\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c9\"\u003e\u003cp\u003e\u003cem\u003emrsH\u003c/em\u003e\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eBH1.0, BH1.1, BH1.2, BH2.1, HT3.0, HT3.1, HT3.2, BT7.0, OK9.0, OK10.0, KK11.2, KK12.2\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\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eBH2.0\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\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eBH2.2\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\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eHT4.0\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\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eHT4.1\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\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eHT4.2\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\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eHT5.0, KK11.3\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\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eHT5.1\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\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eHT5.2\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\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eBT6.0, BT8.2, BT8.3\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\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eBT6.1\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\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eBT6.2\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\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eBT7.1, OK9.1\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\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eBT7.2\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\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eBT8.0\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\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eBT8.1\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\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eOK9.2, KK12.0\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\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eOK9.3, KK11.1\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\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eOK10.1\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\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eOK10.2, KK12.1\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\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eOK10.3\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\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eKK11.0\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\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-\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\u003eHowever, gene associations were noted for some strains. These were BH2.0, HT4.1, HT4.2, HT5.2, and OK10.3, which exhibited coproduction of an antifungal substance composed of the macromolecular proteins flagellin, TasA, and the micromolecular lipopeptide (iturinA).\u003c/p\u003e\u003cp\u003eBased on the results of this study, we detected the catalase-positive, Gram-positive \u003cem\u003eBacillus cereus\u003c/em\u003e in some of our samples. These different \u003cem\u003eBacillus cereus\u003c/em\u003e strains were tested against eight antibiotics \u003cb\u003e(\u003c/b\u003eFig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e; Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e6\u003c/span\u003e\u003cb\u003e)\u003c/b\u003e selected according to EUCAST recommendations.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eThe \u003cem\u003eBacillus\u003c/em\u003e strains exhibited a variable antimicrobial resistance pattern \u003cb\u003e(\u003c/b\u003eTable\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e\u003cb\u003e)\u003c/b\u003e. Indeed, high resistance (100%) was observed against oxacillin and 84.61% resistance against penicillin. At least 65% of \u003cem\u003eBacillus\u003c/em\u003e strains were resistant to amikacin and trimethoprim-sulfamethoxazole. Low resistance was observed against gentamicin (15.38%), erythromycin (30.76%), vancomycin (19.23%), and ciprofloxacin (1%) against sensitivity rates of 84.61%, 69.23%, 80.76%, and 96.15% respectively.\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\u003eSusceptibility of \u003cem\u003eB. cereus\u003c/em\u003e strains to tested antibiotics\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"9\"\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\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eStrains\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colspan=\"8\" nameend=\"c9\" namest=\"c2\"\u003e\u003cp\u003eAntibiotics\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eP\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eOX\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eCN\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eVA\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eSXT\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003eE\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c8\"\u003e\u003cp\u003eAK\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c9\"\u003e\u003cp\u003eCIP\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\u003eBH1.0, BT8.1\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eBH1.1\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eBH1.2\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eBH2.0, BH2.1\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eBH2.2\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eHT3.0, OK10.0\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eHT3.1\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eHT3.2, HT5.0, BT8.2\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eBT6.0, OK9.1\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eBT6.1, BT8.0\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eBT7.0, BT7.1\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eBT8.3, OK9.0, KK11.3\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eOK10.2, KK12.2\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eKK11.2\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eKK12.1\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eR\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eLegend:\u0026nbsp;\u003c/strong\u003e(S), Sensitive; (R), Resistant; (P), Penicillin; (CIP), Ciprofloxacin; (OX), Oxacillin; (CN), Gentamicin; (AK), Amikacin; (E), Erythromycin; (VA), Vancomycin; (SXT), Trimethoprim-sulfamethoxazole.\u0026nbsp;\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eThe genus \u003cem\u003eBacillus\u003c/em\u003e sp., a sporulating, Gram-positive bacterium was detected in honey accordingly to previous findings \u003csup\u003e\u003cspan additionalcitationids=\"CR20\" citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e\u003c/sup\u003e. Moreover, as already reported, the \u003cem\u003eBacillus\u003c/em\u003e isolated from honey in Asia has been described as able to secrete various peptides and bacteriocins with antifungal activities \u003csup\u003e\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e,\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e\u003c/sup\u003e. \u003cem\u003eBacillus\u003c/em\u003e spp. devote a large portion of their genome to secondary metabolism, as an adaptive response to environmental competition. \u003cem\u003eBacillus\u003c/em\u003e spores have been detected in soil and the ocean, where complex microbial communities have been described \u003csup\u003e\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e,\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e\u003c/sup\u003e. By producing secondary metabolites that can inhibit closely related species and other microorganisms in the ecological niche, \u003cem\u003eBacillus\u003c/em\u003e spp. have gained significant survival advantages. In a previous study a variety of secondary metabolites with antibacterial and antifungal properties from \u003cem\u003eBacillus\u003c/em\u003e species, including some non-ribosomal peptides (NRPs) has been found. NRPs are synthesized by non-ribosomal peptide synthetases (NRPSs) and are independent of RNA messenger. NRPs typically undergo extensive modifications, including glycosylation, acylation, and hydroxylation. Due to these modifications, some NRPs are amphiphilic and able to insert into the cell membrane to form pores, such as gramicidin, surfactin, flagellin, iturin, and other lipopeptides.\u003c/p\u003e\u003cp\u003eIn the present study, 6 pairs carrying genes, such as \u003cem\u003eitu\u003c/em\u003eA (IturinA), \u003cem\u003ehag\u003c/em\u003e (Flagellin), \u003cem\u003etas\u003c/em\u003eA (TasA), \u003cem\u003esrf\u003c/em\u003e (Surfactin), \u003cem\u003espa\u003c/em\u003eS (Subtilin) ​​and \u003cem\u003emrs\u003c/em\u003eA (Mersacidin), which are related to the production of antifungal peptides, were investigated. Although the coproduction of three or more antibacterial molecules is not common, two or three lipopeptides simultaneously produced by the same strain have been described. Our results are consistent with those of previous studies \u003csup\u003e\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e\u003c/sup\u003e who identified \u003cem\u003etas\u003c/em\u003eA, \u003cem\u003eitu\u003c/em\u003eA and \u003cem\u003ehag\u003c/em\u003e simultaneously in the same strain of \u003cem\u003eBacillus\u003c/em\u003e sp isolated from honey and demonstrated that these genes conferred properties to control certain fungal species such as \u003cem\u003eFusarium graminearum\u003c/em\u003e, \u003cem\u003eRhizoctonia solani\u003c/em\u003e, and \u003cem\u003eBotrytis cinerea\u003c/em\u003e. According to Zhou and co-workers, the first three antifungal peptides have their own characteristics and antifungal spectrum \u003csup\u003e\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e\u003c/sup\u003e. The iturin family is a cyclic lipopeptide type containing 7 amino acids, with strong antifungal activity. Iturin's antifungal mechanism places its hydrophobic tail into the plasma membrane of indicator cells and automatically assembles to form an ion channel, thus allowing leakage from the cytoplasm.\u003c/p\u003e\u003cp\u003eAdditionally, iturin A can release electrolytes and polymer aggregates, increase the electrical conductivity and permeability of the biomass membrane, resulting in surface tension effects on the cell membrane, inhibiting pathogen formation \u003csup\u003e\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003eThe flagellar filament, formed by a single protein subunit, flagellin, covers 98% of the total length of the flagellum. The flagellin gene is a highly conserved sequence used as a bacterial classification marker. The presence of the gene has been noted in a few strains. According to Nihorimbere and Ongena, lipopeptide production by \u003cem\u003eBacillus\u003c/em\u003e species depends on environmental factors, including temperature, pH, carbon source, and oxygen availability \u003csup\u003e\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e\u003c/sup\u003e. Antifungal lipopeptides produced by \u003cem\u003eBacillus\u003c/em\u003e spp. may have additional applications. These lipopeptides can modify biofilm formation, motility, and virulence gene expression in various microorganisms \u003csup\u003e\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e,\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e\u003c/sup\u003e. The TasA protein is one of these lipopeptides. It is involved in sporulation and biofilm formation in \u003cem\u003eBacillus\u003c/em\u003e \u003csup\u003e\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e\u003c/sup\u003e. These lipopeptides can colonize plant roots, provide plant defenses, and promote plant growth \u003csup\u003e\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e\u003c/sup\u003e. With the increasing need for biopesticides with high specificity, low environmental persistence, and low toxicity, the industrial exploitation of these biological or naturally derived products as food preservatives and plant protection agents is constantly expanding.\u003c/p\u003e\u003cp\u003eOver the past few decades, the incidence of antimicrobial resistance has increased significantly due to the overuse of antibiotics worldwide. According to the results of our study, \u003cem\u003eBacillus cereus\u003c/em\u003e strains isolated from honey exhibited variable antibiotic resistance profiles against the tested molecules. A recent study performed by Noha and co-workers on \u003cem\u003eB. cereus\u003c/em\u003e revealed higher susceptibility than our results against gentamicin (93.3%), ciprofloxacin (86.6%), and vancomycin (83.3%) \u003csup\u003e\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003eHere, such resistance found against beta-lactams could be explained by the large prescription of these antibacterials for infection treatments, and therefore for their misuse. Bacteria have therefore been able to develop natural mechanisms to resist beta-lactams \u003csup\u003e\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e,\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e\u003c/sup\u003e. Generally, \u003cem\u003eBacillus\u003c/em\u003e species secrete beta-lactamase, which hydrolyzes antibiotics \u003csup\u003e\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e,\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e\u003c/sup\u003e of the beta-lactam class, contributing to the development of resistance.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThe objectives that prompted the present study can be considered achieved. First, the microbiological analysis of the honey revealed the presence of \u003cem\u003eBacillus\u003c/em\u003e. Honey produced in Burkina Faso is a valuable source of interesting bacteria with applications in agronomy and the food industry, where an increasing interest in natural, safe, new, and effective solutions against the harmful effects of global warming has been detected. Results revealed that honey represents a valuable matrix for the isolation of probiotic \u003cem\u003eBacillus\u003c/em\u003e strains for interesting technological applications, being able to synthesize up to five peptides with antifungal effects simultaneously, to be proposed in agriculture to ensure sustainable protection of living beings on the planet.\u003c/p\u003e"},{"header":"Declarations","content":"\u003c/p\u003e\u003ch2\u003eFunding\u003c/h2\u003e\u003cp\u003eNo funding has been received for this research as financial support.\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eSHSK contributed to the conceptualization and visualization of the study and contributed to the software analysis of the data. HB handled laboratory manipulations, studied, and wrote most of the paper. UDK contributed to collect and software analysis of data. SS contributed to methodology and collect data during the work. VY, RNTM, RCY, YO, WS, KA, MHD and CC reviewed, corrected, and approved the draft article before submission.\u003c/p\u003e\u003ch2\u003eAcknowledgement\u003c/h2\u003e\u003cp\u003eThe research leading to these results was the result of a collaboration between several entities, including the Applied Biological Sciences Laboratory, the Muraz Center, and the Animal Health and Biotechnology Research and Teaching Laboratory.\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003eYes. The datasets generated and analysed during the current study are available in the [PCR-amplified fragments on 2% agarose gel of Bacillus sp. strains using 16S DNA primers] repository, [Data DOI: 10.57760/sciencedb.28857]\u0026rdquo;.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAbrol, D. P., Shankar, U., Nitin, K. S. \u0026amp; Gowda, G. B. Honeybees and beekeeping: The global scenario. \u003cem\u003eArthropod Divers. Conserv. Tropics Sub-Tropics\u003c/em\u003e. \u003cb\u003e345\u0026ndash;372\u003c/b\u003e \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1007/978-981-10-1518-2_22/TABLES/7\u003c/span\u003e\u003cspan address=\"10.1007/978-981-10-1518-2_22/TABLES/7\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2016).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eJaff\u0026eacute;, R. et al. Bees for development: Brazilian survey reveals how to optimize stingless beekeeping. \u003cem\u003ePLoS One\u003c/em\u003e \u003cb\u003e10\u003c/b\u003e, (2015).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eHill, R. et al. Biocultural approaches to pollinator conservation. \u003cem\u003eNat. Sustain.\u003c/em\u003e \u003cb\u003e2\u003c/b\u003e, 214\u0026ndash;222 (2019).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eVarol, E. \u0026amp; Y\u0026uuml;cel, B. \u003cem\u003eThe Effects of Environmental Problems on Honey Bees in View of Sustainable Life\u003c/em\u003e. \u003cem\u003eMellifera\u003c/em\u003e vol. 19 (2019).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eSawadogo Souha\u0026iuml;bou. Pratiques Apicoles Endog\u0026egrave;nes et Essai de Co-D\u0026eacute;veloppement de Ruches Accessibles Aux Petits Apiculteurs Du Burkina Faso Laboratoire d\u0026rsquo;Entomologie Fondamentale et Appliqu\u0026eacute;e. (2021).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eMoriano, M. E. \u0026amp; Alamprese, C. Honey, trehalose and erythritol as sucrose-alternative sweeteners for artisanal ice cream. A pilot study. \u003cem\u003eLWT\u003c/em\u003e \u003cb\u003e75\u003c/b\u003e, 329\u0026ndash;334 (2017).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAlqarni, A. S. \u0026amp; Honeybee Foraging Nectar Secretion, and Honey Potential of Wild Jujube Trees, Ziziphus nummularia. \u003cem\u003eNeotrop. Entomol.\u003c/em\u003e \u003cb\u003e44\u003c/b\u003e, 232\u0026ndash;241 (2015).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAdgaba, N. et al. Nectar secretion dynamics and honey production potentials of some major honey plants in Saudi Arabia. \u003cem\u003eSaudi J. Biol. Sci.\u003c/em\u003e \u003cb\u003e24\u003c/b\u003e, 180\u0026ndash;191 (2017).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eDua, A., Bahman, S., Kelly, S., Dogra, S. \u0026amp; Sharma, K. Authentication of Indian Honey Based on Carbon Stable Isotope Ratio Analysis\u0026mdash;Verification of Indian Regulatory Criteria. \u003cem\u003eFoods\u003c/em\u003e \u003cb\u003e14\u003c/b\u003e, 1289 (2025).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eFaltus, A., Pawłuszkiewicz, K., Mikołajczak, M. \u0026amp; Korgiel, M. Gaweł-Dąbrowska, D. Benefits of Honey Supplementation for Enhancing Health and Athlete Performance. \u003cem\u003eQual. Sport\u003c/em\u003e. \u003cb\u003e40\u003c/b\u003e, 59354 (2025).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eIkegbunam, N. C., Walter, O. J. \u0026amp; Osayi, E. E. Evaluation of the Floral Diversity and Physicochemical Composition of Honey Procured from some Markets in Anambra State, Nigeria. \u003cem\u003eJ. Appl. Sci. Environ. Manage.\u003c/em\u003e \u003cb\u003e29\u003c/b\u003e, 961\u0026ndash;971 (2025).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eCuvillier \u003cem\u003eMiel, Propolis, Gel\u0026eacute;e Royale: Les Abeilles Alli\u0026eacute;es de Notre Syst\u0026egrave;me Immunitaire.\u003c/em\u003e (2015). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://pharmacie.univ-lille2.fr\u003c/span\u003e\u003cspan address=\"http://pharmacie.univ-lille2.fr\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eInaudi, P. et al. Inorganic Composition as Possible Marker for Botanical and Geological Assignment. \u003cem\u003eMolecules\u003c/em\u003e vol. 30 Preprint at (2025). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/molecules30071466\u003c/span\u003e\u003cspan address=\"10.3390/molecules30071466\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eOgwu, M. C. \u0026amp; Izah, S. C. Honey as a Natural Antimicrobial. \u003cem\u003eAntibiotics\u003c/em\u003e \u003cb\u003e14\u003c/b\u003e, 255 (2025).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAbou Auda, M. M. An updated comprehensive review, systematics and biological activity of medicinal plants used in traditional medecines and their common uses in the Gaza Strip, Palestine. \u003cem\u003eAdv. Environ. Biol.\u003c/em\u003e \u003cb\u003e19\u003c/b\u003e, 1\u0026ndash;43 (2025).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eRoy, S., Mishra, B. K., Pattnaik, M. S. \u0026amp; Mahapatra, S. S. Utilization of Apiary Product for Boosting Human Health. (2025).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAlippi, A. M., Andez, F., Opez, A. C. L. \u0026amp; L. A. \u0026amp; Diversity of aerobic spore-forming bacteria isolated from fresh bee pollen intended for human consumption in Argentina. \u003cem\u003eJ. Apic. Res.\u003c/em\u003e \u003cb\u003e1\u0026ndash;8\u003c/b\u003e \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1080/00218839.2021.1960747\u003c/span\u003e\u003cspan address=\"10.1080/00218839.2021.1960747\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2021).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAlippi, A. M., Fern\u0026aacute;ndez, L. A. \u0026amp; L\u0026oacute;pez, A. C. Diversity of aerobic spore-forming bacteria isolated from fresh bee pollen intended for human consumption in Argentina. \u003cem\u003eJ. Apic. Res.\u003c/em\u003e \u003cb\u003e61\u003c/b\u003e, 392\u0026ndash;399 (2022).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eBł\u0026oacute;nska, D. \u0026amp; Buszewski, B. Characterization of Honey Microbiome Using MALDI-TOF Mass Spectrometry and Physicochemical Study. (2025). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3390/molecules30061266\u003c/span\u003e\u003cspan address=\"10.3390/molecules30061266\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eCirtwill, A. R. \u0026amp; Wirta, H. DNA in honey could describe the changes in flower visits and microbe encounters of honey bees over decades. \u003cem\u003eSci. Rep.\u003c/em\u003e \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1038/s41598-025-93315-8\u003c/span\u003e\u003cspan address=\"10.1038/s41598-025-93315-8\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2025).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eWang, M., Zhao, W., Zhou, D. \u0026amp; Huang, J. Bacterial diversity in the honey sac during bee foraging on winter-flowering flora and dominant Bacillus subtilis inhibits Hafnia alvei. \u003cem\u003eFront. Insect Sci.\u003c/em\u003e \u003cb\u003e5\u003c/b\u003e, 1555434 (2025).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eISO 6887-1. International Organization for Standardization. No.6887-1. Microbiology of the Food Chain \u0026mdash; Preparation of Test Samples, Initial Suspension and Decimal Dilutions for Microbiological Examination \u0026mdash; Part 1: General Rules for the Preparation of the Initial Suspension and Decimal Dilutions.. (2017).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eOuoba, L. I. I., Parkouda, C., Diawara, B., Scotti, C. \u0026amp; Varnam, A. H. Identification of Bacillus spp. from Bikalga, fermented seeds of Hibiscus sabdariffa: Phenotypic and genotypic characterization. \u003cem\u003eJ. Appl. Microbiol.\u003c/em\u003e \u003cb\u003e104\u003c/b\u003e, 122\u0026ndash;131 (2008).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eGounina-Allouane, R., Broussolle, V. \u0026amp; Carlin, F. Influence of the sporulation temperature on the impact of the nutrients inosine and L-alanine on Bacillus cereus spore germination. \u003cem\u003eFood Microbiol.\u003c/em\u003e \u003cb\u003e25\u003c/b\u003e, 202\u0026ndash;206 (2008).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eSusilowati, R., Sabdono, A. \u0026amp; Widowati, I. Isolation and Characterization of Bacteria Associated with Brown Algae Sargassum spp. from Panjang Island and their Antibacterial Activities. \u003cem\u003eProcedia Environ. Sci.\u003c/em\u003e \u003cb\u003e23\u003c/b\u003e, 240\u0026ndash;246 (2015).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eZhou, Z. et al. Investigation of antibacterial activity of Bacillus spp. isolated from the feces of Giant Panda and characterization of their antimicrobial gene distributions. \u003cem\u003eWorld J. Microbiol. Biotechnol.\u003c/em\u003e \u003cb\u003e30\u003c/b\u003e, 3129\u0026ndash;3136 (2014).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eCASFM. Comit\u0026eacute; de l \u0026rsquo; antibiogramme de la Soci\u0026eacute;t\u0026eacute; Fran\u0026ccedil;aise de Microbiologie Recommandations 2022. (2022).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eEUCAST. Bacillus spp. Calibration of zone diameter breakpoints to MIC values. (2023). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.eucast.org/fileadmin/src/media/PDFs/EUCAST_files/Disk_criteria/Validation_2023/Bacillus_v_2.1_February_2023.\u003c/span\u003e\u003cspan address=\"https://www.eucast.org/fileadmin/src/media/PDFs/EUCAST_files/Disk_criteria/Validation_2023/Bacillus_v_2.1_February_2023.\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003cem\u003epdf\u003c/em\u003e 1\u0026ndash;17.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eMills, E., Sullivan, E. \u0026amp; Kovac, J. Comparative Analysis of Bacillus cereus Group Isolates\u0026rsquo; Resistance Using Disk Diffusion and Broth Microdilution and the Correlation between Antimicrobial Resistance Phenotypes and Genotypes. (2022).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eZhao, X. et al. Isolation and identification of antifungal peptides from bacillus BH072, a novel bacterium isolated from honey. \u003cem\u003eMicrobiol. Res.\u003c/em\u003e \u003cb\u003e168\u003c/b\u003e, 598\u0026ndash;606 (2013).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eChen, M., Takano, C., Nakashima, K., Gowthaman, S. \u0026amp; Kawasaki, S. Exploration of ureolytic airborne bacteria for biocementation applications from different climate zones in Japan. \u003cem\u003eSci. Rep.\u003c/em\u003e \u003cb\u003e15\u003c/b\u003e, 7536 (2025).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAl-Abdulsalam, H., Almalki, M. \u0026amp; Khalifa, A. Isolation and characterization of antimicrobial metabolites producing bacteria from soils in Al-Ahsa, Saudi Arabia. \u003cem\u003eJournal Microbiol. Biotechnol. Food Sciences\u003c/em\u003e \u003cb\u003e14\u003c/b\u003e, (2025).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eMarkelova, N. \u0026amp; Chumak, A. Antimicrobial Activity of Bacillus Cyclic Lipopeptides and Their Role in the Host Adaptive Response to Changes in Environmental Conditions. \u003cem\u003eInt. J. Mol. Sci.\u003c/em\u003e \u003cb\u003e26\u003c/b\u003e, 336 (2025).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eNihorimbere, V. \u0026amp; Ongena, M. Isolation of Plant growth-promoting Bacillus Strains with Biocontrol Activity In vitro. \u003cem\u003eMerit Res. J. Microbiol. Biol. Sci.\u003c/em\u003e \u003cb\u003e5\u003c/b\u003e, 2408\u0026ndash;7076 (2017).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eKumar, M. et al. Unraveling the Bioactivities and Immunomodulatory Potential of Postbiotics Derived from Bacillus subtilis and B. amyloliquefaciens for Aquaculture. \u003cem\u003eProbiotics Antimicrob. Proteins\u003c/em\u003e \u003cb\u003e1\u0026ndash;15\u003c/b\u003e (2025).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eRoque-Borda, C. A. et al. Antimicrobial Peptides: A Promising Alternative to Conventional Antimicrobials for Combating Polymicrobial Biofilms. \u003cem\u003eAdv. Sci.\u003c/em\u003e \u003cb\u003e12\u003c/b\u003e, 2410893 (2025).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eKim et al. Production of Biosurfactant Lipopeptides Iturin A, Fengycin, and Surfactin A from Bacillus subtilis CMB32 for Control of Colletotrichum gloeosporioides.pdf. Preprint at (2010).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eNoha et al. Identification, characterization and antibiotic susceptibility testing for Bacillus species. \u003cb\u003e14\u003c/b\u003e, 484\u0026ndash;494 (2022).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eElshobary, M. E. et al. Combating Antibiotic Resistance: Mechanisms, Multidrug-Resistant Pathogens, and Novel Therapeutic Approaches: An Updated Review. \u003cem\u003ePharmaceuticals\u003c/em\u003e \u003cb\u003e18\u003c/b\u003e, 402 (2025).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eVivekanandan, K. E., Kumar, P. V., Jaysree, R. C. \u0026amp; Rajeshwari, T. Exploring Molecular Mechanisms of Drug Resistance in Bacteria and Progressions in CRISPR/Cas9-Based Genome Expurgation Solutions. \u003cem\u003eGlob Med. Genet\u003c/em\u003e \u003cb\u003e100042\u003c/b\u003e (2025).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAlemayehu, T. et al. Phenotypic identification of Metallo-\u0026szlig;-lactamase resistance Gram negative bacteria from a clinical specimen in Sidama, Ethiopia. \u003cem\u003ePLoS One\u003c/em\u003e. \u003cb\u003e20\u003c/b\u003e, e0313431 (2025).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eGeremia, N. et al. Rare or Unusual Non-Fermenting Gram-Negative Bacteria: Therapeutic Approach and Antibiotic Treatment Options. \u003cem\u003eAntibiotics\u003c/em\u003e \u003cb\u003e14\u003c/b\u003e, 306 (2025).\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Honey, Burkina Faso, Bacillus sp., genetic diversity, RT-PCR, antimicrobial peptides","lastPublishedDoi":"10.21203/rs.3.rs-7280305/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7280305/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground: \u003c/strong\u003eBees and honey are garnering attention as potential sources for isolating beneficial microorganisms. However, the diversity of antibacterial peptides from microorganisms in honey produced in Burkina Faso remains poorly documented. This study aims to explore the diversity of genes detected in \u003cem\u003eBacillus\u003c/em\u003e sp. isolated from honey in Burkina Faso.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults:\u003c/strong\u003e Here 11 honey samples were used as a matrix to isolate \u003cem\u003eBacillus\u003c/em\u003espp. bacteria. A total of 40 presumptive \u003cem\u003eBacillus\u003c/em\u003e spp. isolates, preliminarily characterized for Gram staining, catalase tests, and survival under thermal stress, were selected. The RT-PCR with universal 16S primers revealed that all 40 isolates belonged to the genus \u003cem\u003eBacillus \u003c/em\u003eand the presence of genes involved in iturine A, flagellin, TasA, surfactin, subtilin, and mersacidin synthesis. Among the isolates, 65% carry the \u003cem\u003eBacillus\u003c/em\u003e \u003cem\u003ecereus\u003c/em\u003egene. The rest consists (about 40%) of beneficial strains possessing at least one out of six genes and five strains exhibited at least three genes simultaneously. Associations of more than three genes have detected in \u003cem\u003eBacillus\u003c/em\u003e sp. strains, however they are almost absent in \u003cem\u003eBacillus cereus\u003c/em\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion:\u003c/strong\u003e Honey can be considered a natural source of beneficial bacteria belonging to \u003cem\u003eBacillus\u003c/em\u003e genus with a broad spectrum of antimicrobial activities useful for plant bioprotection applications.\u003c/p\u003e","manuscriptTitle":"Unveiling the culturable Bacillus sp. from honey in Burkina Faso as a potential source for antimicrobial peptides in plant disease control","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-08-22 07:07:33","doi":"10.21203/rs.3.rs-7280305/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"0ce75d0f-45e7-4e04-9eff-aa87bc1c3c31","owner":[],"postedDate":"August 22nd, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":53550267,"name":"Biological sciences/Biotechnology"},{"id":53550268,"name":"Biological sciences/Microbiology"},{"id":53550269,"name":"Biological sciences/Molecular biology"}],"tags":[],"updatedAt":"2026-03-02T07:11:10+00:00","versionOfRecord":[],"versionCreatedAt":"2025-08-22 07:07:33","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7280305","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7280305","identity":"rs-7280305","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}