Bacteriocinogenic Potential Research Articles

Bacteriocinogenic anti-listerial properties and safety assessment of Enterococcus faecium and Lactococcus garvieae strains isolated from Brazilian artisanal cheesemaking environment.

This study aimed to prospect and isolate lactic acid bacteria (LAB) from an artisanal cheese production environment, to assess their safety, and to explore their bacteriocinogenic potential against Listeria monocytogenes. Samples were collected from surfaces of an artisanal-cheese production facility and after rep-PCR and 16S rRNA sequencing analysis, selected strains were identified as to be belonging to Lactococcus garvieae (1 strain) and Enterococcus faecium (14 isolates, grouped into three clusters) associated with different environments (worktables, cheese mold, ripening wooden shelves). All of them presented bacteriocinogenic potential against L. monocytogenes ATCC 7644 and were confirmed as safe (γ-hemolytic, not presenting antibiotic resistance, no mucus degradation properties, and no proteolytic or gelatinase enzyme activity). Additionally, cell growth, acidification and bacteriocins production kinetics, bacteriocin stability in relation to different temperatures, pH, and chemicals were evaluated. According to performed PCR analysis all studied strains generated positive evidence for the presence of entA and entP genes (for production of enterocins A and enterocins P, respectively). However, pediocin PA-1 associated gene was recorded only in DNA obtained from E. faecium ST02JL and Lc. garvieae ST04JL. It is worth considering the application of these safe LAB or their bacteriocins in situ as an alternative means of controlling L. monocytogenes in cheese production environments, either alone or in combination with other antimicrobials.

IN VITRO evaluation of probiotic properties and bacteriocinogenic potential of strains of microorganisms L. Acidophilus 13, E. Faecium K-50 and their compositions

It is known that the gut microbiota plays a central role in the health and biology of the animal, and is therefore important for scientific research. Microorganisms of the genus Lactobacllus are widespread in nature and are considered representatives of the microflora of the macroorganism of the animal. Lactobacllus acidophilus is widely used as a probiotic, and is known for its immunostimulating effect, it colonizes the intestines of a newborn and accompanies the animal throughout its life. Enterococcus faecium is also used as a probiotic, due to its high antagonistic activity against pathogenic microbiota, and also participates in the formation and maintenance of immunity. The purpose of this study was to evaluate the probiotic component of L. acidophilus 13, E. faecium k-50 in conditions simulating the gastrointestinal tract, to study the bacteriocinogenic potential and antagonistic properties of lactobacilli and enterococci and their composition. In this review, we summarize recent advances in understanding the mechanisms involved in bacterial tolerance to bile, with particular emphasis on research contributions. In addition, the physiological and ecological role of enzymes and their importance for animal health, as well as the function of bile acid as a regulator of metabolism, are discussed. Samples were used in the study.

Bioactivity Screening and Genomic Analysis Reveals Deep-Sea Fish Microbiome Isolates as Sources of Novel Antimicrobials.

With the increase in antimicrobial resistance and the subsequent demand for novel therapeutics, the deep-sea fish microbiome can be a relatively untapped source of antimicrobials, including bacteriocins. Previously, bacterial isolates were recovered from the gut of deep-sea fish sampled from the Atlantic Ocean.In this study, we used in vitro methods to screen a subset of these isolates for antimicrobial activity, and subsequently mined genomic DNA from isolates of interest for bacteriocin and other antimicrobial metabolite genes. We observed antimicrobial activity against foodborne pathogens, including Staphylococcus aureus, Listeria monocytogenes, Enterococcus faecalis and Micrococcus luteus. In total, 147 candidate biosynthetic gene clusters were identified in the genomic sequences, including 35 bacteriocin/RiPP-like clusters. Other bioactive metabolite genes detected included non-ribosomal peptide synthases (NRPS), polyketide synthases (PKS; Types 1 and 3), beta-lactones and terpenes. Moreover, four unique bacteriocin gene clusters were annotated and shown to encode novel peptides: a class IIc bacteriocin, two class IId bacteriocins and a class I lanthipeptide (LanM subgroup). Our dual in vitro and in silico approach allowed for a more comprehensive understanding of the bacteriocinogenic potential of these deep-sea isolates and an insight into the antimicrobial molecules that they may produce.

Genomic insights into the safety and bacteriocinogenic potential of isolates from artisanal fermented milk Matsoni

Matsoni is a fermented milk common in the Caucasian region. In this study, whole-genome sequencing was performed with the aim of determining the taxonomic position of 25 previously selected candidate Matsoni starter cultures, identifying bacteriocin genes and prophages in their genomes, and evaluating their safety. In silico prediction of antibiotic resistance genes, virulence factors, bacteriocin genes and prophages was performed using dedicated databases and bioinformatics tools. With the exception of Enterococcus faecalis, which contained the mobile genetic element Tn916 with tetM and virulence factors, and Streptococcus thermophilus with mefE, the isolates were found to be safe for use as starter cultures. Thermophilin A, thermophilin 13, streptide and Blp genes were predicted in isolates of Streptococcus thermophilus, and genes for enterocins A, B, and X were found in Enterococcus faecium, but inhibition between strains was not observed. Several complete prophages were also detected in the genomes of Lactobacillus delbrueckii ssp. lactis. Lacticaseibacillus rhamnosus, Limosilactobacillus fermentum, Streptococcus thermophilus, Enterococcus faecalis, and Enterococcus faecium. Some of these isolates are good candidates for customised autochthonous Matsoni starter cultures which are an alternative to the commercial starter cultures currently used in the industry.

Genomic, metabolomic, and functional characterisation of beneficial properties of Pediococcus pentosaceus ST58, isolated from human oral cavity.

Bacteriocins produced by lactic acid bacteria are proteinaceous antibacterial metabolites that normally exhibit bactericidal or bacteriostatic activity against genetically closely related bacteria. In this work, the bacteriocinogenic potential of Pediococcus pentosaceus strain ST58, isolated from oral cavity of a healthy volunteer was evaluated. To better understand the biological role of this strain, its technological and safety traits were deeply investigated through a combined approach considering physiological, metabolomic and genomic properties. Three out of 14 colonies generating inhibition zones were confirmed to be bacteriocin producers and, according to repPCR and RAPD-PCR, differentiation assays, and 16S rRNA sequencing it was confirmed to be replicates of the same strain, identified as P. pentosaceus, named ST58. Based on multiple isolation of the same strain (P. pentosaceus ST58) over the 26 weeks in screening process for the potential bacteriocinogenic strains from the oral cavity of the same volunteer, strain ST58 can be considered a persistent component of oral cavity microbiota. Genomic analysis of P. pentosaceus ST58 revealed the presence of operons encoding for bacteriocins pediocin PA-1 and penocin A. The produced bacteriocin(s) inhibited the growth of Listeria monocytogenes, Enterococcus spp. and some Lactobacillus spp. used to determine the activity spectrum. The highest levels of production (6400 AU/ml) were recorded against L. monocytogenes strains after 24 h of incubation and the antimicrobial activity was inhibited after treatment of the cell-free supernatants with proteolytic enzymes. Noteworthy, P. pentosaceus ST58 also presented antifungal activity and key metabolites potentially involved in these properties were identified. Overall, this strain can be of great biotechnological interest towards the development of effective bio-preservation cultures as well as potential health promoting microbes.

Safety Assessment of Fecal, Bacteriocin-Producing Strains Enterococcus mundtii from Horses

Backround: Knowing and optimizing the host microbiota is important regarding the maintenance of horse`s health. For this approach beneficial bacteria have been usually used. However, to be the most effective in use, their safety, tolerability and efficacy needs to be assessed. Therefore, fecal strains Enterococcus mundtii from horses with promising bacteriocinogenic potential have to be evaluated for their safety. Methodology: E. mundtii strains were isolated from feces of 47 horses (n=47; 40 mares and seven stallions), the Norik breed from Muráň in eastern Slovakia. MALDTI-TOF spectrometry and sequencing were used. Phenotypic characteristics were assessed in accordance with those for the reference strain E. mundtii ATCC43186. Biofilm-forming ability was tested using plate assay. For enzyme production commercial tests were applied and virulence factor genes were tested using PCR and primers. Antibiotic profile was tested with diffusion method. In vivo safety was tested using Balb/c breed mice. Results: E. mundtii strains did not produce the enzyme β-glucuronidase; however, most E. mundtii strains produced β-galactosidase. The strains were absent of virulence factors such as gelatinase, aggregation substance, cytolysin A, enterococcal superficial protein, adhesins, hyaluronidase and IS16 element, except efaAfs gene in the strain EMKD 24/1. Six strains were found with lowgrade (0.1 ≤ A570< 1) biofilm - forming ability. E. mundtii were mostly susceptible to antibiotics. Bacteriocinogenic strain EMKD41/3 reached high counts in feces (5.12 ± 0.26 CFU/g log 10) of Balb/c mice during its 30 days application. No mortality of mice was noted during whole period of EMKD41/3 strain application. Conclusion: Bacteriocin-producing strains E. mundtii should not threaten horses because they were mostly susceptible to antibiotics, they were virulence factor genes absent and with low-grade biofilm formation ability. Bacteriocinproducing strain EMKD41/3 even indicates its sufficient implementation in host organism with any side effect.

Natural Bioactive Compounds in Organic and Conventional Fermented Food.

Compared to conventional agriculture, organic farming is believed to provide a higher nutritional and health value in its products due to the elimination of harmful contaminants (pesticides, nitrates, heavy metals, etc.). Numerous studies have been conducted to show how the production system affects the quality of food in terms of the content of bioactive compounds. The aim of this study was to compare the content of some bioactive compounds (vitamin C, β-carotene, Ca content) and lactic acid bacteria (LAB) number and their bacteriocinogenic activity in organic and conventional fermented food. Although the results do not provide an unambiguous conclusion regarding the superiority of one production system over the other, the LAB number in organic pickled carrot juice, sauerkraut, yogurt, and kefir was higher than in their conventional counterparts. Their bacteriocinogenic potential against selected pathogens was also higher in most organic products. Organic vegetables contained significantly more vitamin C, and the calcium content in the organic yogurt was higher compared to the conventional version of the product. Relatively similar concentrations of ß-carotene for both production systems were found in carrot juice, while in organic pickled beet juice, there was five-fold less ß-carotene than in conventional juice.

Effects of lactic acid bacteria with bacteriocinogenic potential on the chemical composition and fermentation profile of forage peanut (Arachis pintoi) silage

The objectives of this study were to evaluate the effect of lactic acid bacteria (LAB) with bacteriocinogenic activity isolated from stylo silages on the microbial population, chemical composition and fermentation profile of forage peanut silage (FPS) and to identify LAB species in forage peanut and its silages based on the 16 S rRNA sequence analysis. The experiment was conducted in a completely randomized design with a 5 × 6 factorial arrangement (five inoculants (I) × six fermentation periods (P)) and three replicates. The following inoculants were evaluated: without inoculant (CTRL); a commercial inoculant at a dose of 107 colony forming units (cfu) g-1 fresh matter (CI1); a commercial inoculant at a dose of 105 cfu g-1 fresh matter (CI2); Pediococcus acidilactici strain 10.6 (S1); and Pool of bacteriocinogenic strains (POOL), including S1, P. pentosaceus strain 6.16 and P. acidilactici strain 10.4. The fermentation periods were 1, 3, 7, 14, 28 and 56 days. The largest LAB populations were observed in silages treated with S1 (8.66 log cfu g-1) and POOL (8.69 log cfu g-1). At the end of the fermentation period, a higher content of dry matter (222.53 g kg-1) and lactic acid (66.30 g kg-1) and lower values of pH (4.76) and acid detergent insoluble nitrogen (9.50% of total nitrogen) were observed for silage treated with S1. The screening of LAB from forage peanut silage revealed that P. pentosaceus was the predominant species. In addition, the wild strain of P. acidilactici 10.6 established a desirable microbial population during the fermentation process reflecting a highest lactic acid concentration and lowest pH values after 56 days of fermentation.

Bacteriocinogenic Potential of Bacillus amyloliquefaciens Isolated from Kimchi, a Traditional Korean Fermented Cabbage.

Bacteriocin production is considered a favorable property for various beneficial cultures. In addition to their potential as biopreservatives, bacteriocins are also promising alternatives for the control of multidrug-resistant pathogens and the inhibition of some viruses and cancer cells. The objective of this study was to screen and characterize a bacteriocin-producing strain with the aim of its future application for control of Listeria monocytogenes, an important food-borne pathogen. A total of 22 potentially bacteriocinogenic strains active against L. monocytogenes ATCC15313 were isolated from locally produced kimchi through a three-level approach. Pure cultures were obtained according to good microbiological practices and differentiated through RAPD-PCR using the primers OPL01, OPL09, and OPL11. Altogether, 5 strains were selected for further study. Specific focus was given to strain ST05DL based on its specific inhibitory activity against L. monocytogenes ATCC15313, while not affecting different strains belonging to the genera Lactobacillus, Pediococcus, Leuconostoc, and Weissella, most of which are beneficial microorganisms. The strain ST05DL was identified as Bacillus amyloliquefaciens based on its sugar fermentation profile obtained through API50CHB analysis and 16S rRNA partial sequencing. The antimicrobial compound produced by B. amyloliquefaciens ST05DL was found to be sensitive to pepsin and α-chymotrypsin, evidence of its proteinaceous nature. The presence of skim milk, NaCl, Tween 80, glycerol, and SDS did not affect the antimicrobial activity. The addition of 20% cell-free supernatant (CFS) obtained from a 24-h culture of B. amyloliquefaciens ST05DL to an exponentially growing culture of L. monocytogenes ATCC15313 successfully inhibited the test microorganisms during the monitored 10-h incubation. Optimal bacteriocin production by B. amyloliquefaciens ST05DL was observed during the stationary phase at 12 h (800 AU/mL) and remained stable for the next 15 h. The ratio between live and dead cells during this period was 74.37% and 25.66%, respectively, as determined by flow cytometry. The presence of the virulence genes hblA, hblB, hblC, nheA, nheB, and nheC was not detected in the total DNA of B. amyloliquefaciens ST05DL, and the strain was resistant only to ampicillin out of 10 tested antibiotics. Future evaluation of expressed bacteriocin/s by B. amyloliquefaciens ST05DL (amino acid sequence, molecular mass, cytotoxicity, detailed mode of action, etc.), will be the next step in the characterization and its potential application as biopreservative and/or pharmaceutical product.

Enterococcus mundtii Isolated from Slovak Raw Goat Milk and Its Bacteriocinogenic Potential.

Enterococci are lactic acid bacteria. Most of them can adapt well to the food system due to their salt and acid-tolerance. Moreover, many enterococcal species have been found to produce antimicrobial substances of proteinaceous character, i.e., bacteriocins/enterocins. In this study, Enterococcus mundtii EM ML2/2 with bacteriocinogenic potential was identified in Slovak raw goat milk. This strain demonstrated inhibition activity against up to 36% of Gram-positive indicator bacteria, and in concentrated form the bacteriocin substance (pH 6.3) showed the highest inhibition activity (1600 AU/mL) against the principal indicator strain E. avium EA5. Semi-purified substance (SPS) EM ML2/2 produced inhibition activity up to 3200 AU/mL. Concentrated bacteriocin substance and SPS maintained active (inhibition activity up to 100 AU/mL) for three months under −20 °C storage conditions. The strain showed susceptible antibiotic profile, and it did not form biofilm. No production of damaging enzymes was noted. It was nonhemolytic, as well as DNase, and gelatinase-negative. It grew well in skim milk, and it was salt and acid-tolerant. The bacteriocin potential of E. mundtii species isolated from Slovak raw goat milk has not previously been detected, so this is an original contribution which may stimulate addtitional research and application studies.

Safety evaluation and bacteriocinogenic potential of Pediococcus acidilactici strains isolated from artisanal cheeses

Pediococcus acidilactici ST1607V, ST2104V and ST3105V, isolated from a Brazilian artisanal cheese expressed bacteriocins with high inhibitory activity against several strains of Listeria monocytogenes and Enterococcus spp., but not against most of the other tested lactic acid bacteria (LAB). Bacteriocinogenic activity against L. monocytogenes ATCC7644 was expressed with a high titre of 51200 AU/mL (ST1607V and ST2104V) and 25600 AU/mL (ST3105V), when cultured in MRS. Bacteriocins produced by strains ST1607V, ST2104V and ST3105V presented a bactericidal mode of action against L. monocytogenes ATCC7644 and Enterococcus faecium ATCC19434. DNA of the studied LAB strains was obtained and subjected to PCR assays targeting more than 50 genes related to biogenic amines production (histidine, tyrosine and ornithine decarboxylases), virulence (sex pheromones, gelatinase, cytolysin, hyaluronidase, aggregation substance, enterococcal surface protein, endocarditis antigen, adhesion of collagen, integration factors) and antibiotic resistance (vancomycin, tetracycline, erythromycin, gentamicin, chloramphenicol, bacitracin). A few safety-related genes were detected in some strains, indicating their safety for application in fermented food products. Special attention should be given to the presence of possible virulence factors, in addition to the production of biogenic amines and antibiotic resistance. Additional in vitro and in vivo experiments could be designed for future evaluation of possible expression of detected virulence genes.

Pastırmadaki Potansiyel Bakteriyosinojenik Laktik Asit Bakterileri

Recent researches conducted have focused on the possibility of using bacteriocin produced by lactic acid bacteria both (LAB) as protective cultures and as natural antimicrobial agents in dry cured meat products such as pastirma. In the present study, 50 lactic acid bacteria strains, previously isolated from various traditional pastirma samples, were tested for their antimicrobial potentials. Determination of antagonistic activities against food-borne pathogenic strains (Listeria monocytogenes, Staphylococcus aureus and Bacillus cereus) were done by using agar-spot and well diffusion assays. Then, active isolates were identified by using 16S rRNA gene sequence analysis. According to the results, 11 of tested LAB were determined with significant bacteriocinogenic potential and these were assigned to as 1 Lactobacillus plantarum, 9 L. plantarum subsp. plantarum and 1 Pediococcus pentosaceus. In conclusion, it was reached that the isolated/identified bacteriocin-producing lactic acid bacteria strains from pastirma samples have a significant potential to prepare pure or crude bacteriocin preparations.

Exploring Beneficial/Virulence Properties of Two Dairy-Related Strains of Streptococcus infantarius subsp. infantarius.

The genus Streptococcus includes various species, remarkably different in their behavior, applications, virulence, and safety. Taxonomically Streptococcus infantarius subsp. infantarius belonging to the Streptococcus bovis group, which includes several pathogen species, however, has been found as predominant species in some African dairy products that are widely consumed and considered to be safe. Streptococcus infantarius subsp. infantarius' safety may be questioned due to the association of this species with clinical cases. In this study, isolates from dairy origin were selected based on their bacteriocinogenic potential and differentiated by their RAPD-PCR profiles. Two strains were identified by 16S rRNA sequencing as St. infantarius subsp. infantarius and investigated regarding their potential beneficial properties and factors related to virulence and safety. A series of in vitro tests included properties related to survival in the gastrointestinal tract and beneficial intestinal activities. Production of bacteriocin/s, detection of related genes, and partial characterization of expressed antimicrobial protein were evaluated. Genes related to folate biosynthesis were detected in both studied strains. Evaluation of physiological tests related to strains virulence, adhesion, and resistance to antibiotics and detections of virulence and biogenic amines production-related genes were also investigated. Taking in consideration all the aspects of the specific nature of St. infantarius subsp. infantarius K1-4 and K5-1 (beneficial properties and virulence characteristics), both strains cannot be considered safe for human or other animals application, even though they have been isolated from dairy products. This study is highlighting the importance of evaluation for presence of potential virulence factors in newly characterized strains in order to be confident in their safety.

The ability of Lactococcus lactis subsp. lactis bv. diacetylactis strains in producing nisin.

Lactococcus lactis subsp. lactis bv. diacetylactis is a relevant microorganism for the dairy industry because of its role in the production of aromatic compounds. Despite this technological property, the identification of bacteriocinogenic potential of obtained strains can offer the additional positive aspect of biosafety. A panel of 15 L. lactis subsp. lactis bv. diacetylactis strains was characterised for the presence and expression of bacteriocin related genes, and further investigated regarding the nisin operon. Eight strains were positive only for nisA, and one strain (SBR4) presented a full nisin operon, with sequencing that was shown to be similar to nisin Z. Only SBR4 presented inhibitory activity against 16 microbial target strains. The growth curves of selected targets strains confirmed the inhibitory activity of SBR4 and consequently the nisin production. This research has demonstrated the inhibitory potential of L. lactis subsp. lactis bv. diacetylactis strain, SBR4, due to its ability to produce nisin Z. This biopreservative potential, associated to previously characterised technological properties, allow the indication of this strain as a promising candidate to be used by the dairy industry as a starter or adjunct culture.

Search and Selection of Bacteriocin-Producing Strains of Marine Bacteria in the Aquatic Ecosystems of the Sea of Japan

Given the increasing antibiotic resistance of pathogenic bacteria, alternative antimicrobial strategies are being investigated. One promising area is the study of the antibacterial activity of bacteriocins, special peptides produced by bacteria, which play an important role in regulating the competitive inter- and intraspecific relationships in natural microbial systems. Marine microorganisms are a rich and promising source of bacteriocins. Marine ecosystems contain a huge variety of organisms, up to 90% of which are bacteria living in highly competitive and extreme conditions. Isolation and study of the antibacterial activity of bacteriocins of marine origin is a promising and unexplored area of innovative biotechnology. This article presents the results of a pilot study on the search and selection of bacteriocin-producing strains of marine bacteria from ecosystems of the Sea of Japan. Pseudoalteromonas elyakovii, Cobetia amphilecti, Paraglaciecola mesophila, and Idiomarina abyssalis clusters contain genes similar to Escherichia coli K12: J62 DNA, which are involved in the biosynthesis of colicins (microcin V MccV). This suggests a bacteriocinogenic potential of the studied strains of marine bacteria. The subsequent isolation and study of biological activity of these peptides will make it possible to draw a conclusion about the biological properties and mechanisms of the antagonistic action of these important compounds and the prospects for their use in industrial breeding of seafood (mariculture), as well as for medical and veterinary purposes.

Preservation of Meat Products with Bacteriocins Produced by Lactic Acid Bacteria Isolated from Meat

Bacteriocins are ribosomal-synthesized antimicrobial peptides that inhibit the growing of pathogenic and/or deteriorating bacteria. The most studied bacteriocin-producing microorganisms are lactic acid bacteria (LAB), as they have great potential application in food biopreservation, since the majority have GRAS (Generally Recognized as Safe) status. The LAB-producing bacteriocins and/or bacteriocins produced by these bacteria have been widely studied, with the emphasis on those derived from milk and dairy products. On the other hand, isolates from meat and meat products are less studied. The objective of this review is to address the main characteristics, classification, and mechanism of action of bacteriocins and their use in food, to highlight studies on the isolation of LAB with bacteriocinogenic potential from meat and meat products and also to characterize, purify, and apply these bacteriocins in meat products. In summary, most of the microorganisms studied areLactococcus,Enterococcus,Pediococcus, andLactobacillus, which produce bacteriocins such as nisin, enterocin, pediocin, pentocin, and sakacin, many with the potential for use in food biopreservation.

In vitro Assessment of the Probiotic Properties and Bacteriocinogenic Potential of Pediococcus pentosaceus MZF16 Isolated From Artisanal Tunisian Meat "Dried Ossban".

Pediococcus pentosaceus MZF16 has been isolated from artisanal Tunisian meat so called “Dried Ossban,” an original ecological niche, and identified by MALDI-TOF mass spectrometry and 16S rDNA sequencing. This bacterium showed a high tolerance to gastric stress conditions, and toward bile salts. P. pentosaceus MZF16 also demonstrated a hydrophobic surface profile (high adhesion to xylene), autoaggregation, and adhesive abilities to the human intestinal Caco-2/TC7 cell line. These properties may help the bacterium colonizing the gut. Furthermore, MZF16 was found to be resistant to gentamycin and chloramphenicol but did not harbor any transferable resistance determinants and/or virulence genes. The data also demonstrated absence of cytotoxicity of this strain. Conversely, P. pentosaceus MZF16 can slightly stimulate the immune system and enhance the intestinal epithelial barrier function. Moreover, this bacterium has been shown to be highly active against Listeria spp. due to bacteriocin production. Characterization of the bacteriocin by PCR amplification, sequencing and bioinformatic analyses revealed that MZF16 produces a bacteriocin 100% identical to coagulin, a pediocin-like inhibitory substance produced by Bacillus coagulans. To our knowledge, this is the first report that highlights the production of a pediocin 100% identical to coagulin in a Pediococcus strain. As coagulin, pediocin MZF16 has the consensus sequence YYGNGVXCXXXXCXVXXXXA (X denotes any amino acid), which confirms its belonging to class IIa bacteriocins, and its suitability to preserve foods from Listeria monocytogenes development. According to these results, P. pentosaceus MZF16 can be proposed as a probiotic and bioprotective agent for fermented foods, including Tunisian dry meat and sausages. Further investigations will aim to study the behavior of this strain in meat products as a component of functional food.

Bacteriocinogenic properties of Escherichia coli P2C isolated from pig gastrointestinal tract: purification and characterization of microcin V.

The aim of this study was to isolate and investigate the bacteriocinogenic and probiotic potential of new Gram-negative isolates. Of 22 bacterial isolates from pig intestine and chicken crops, ten isolates had demonstrated a good activity, and the most potent five strains were identified as four E. coli and one as Proteus sp. No virulence factors were detected for E. coli strains isolated from pig intestine. The semi-purified microcins proved to be resistant to temperature and pH variation, but sensitive to proteolytic enzymes. Of particular interest, strain E. coli P2C was the most potent, free of virulence genes and sensitive to tested antibiotics. Purification procedure revealed the presence of a single pure peak having a molecular mass of 8733.94Da and matching microcin V (MccV). The sequence obtained by LC-MS/MS confirmed the presence of MccV. Purified MccV showed a good activity against pathogenic coliforms, especially E. coli O1K1H7 involved in avian colibacillosis. The present study provides evidence that E. coli strains isolated from pig intestine produce microcin-like substances. E. coli P2C is a safe MccV producer that could be a good candidate for its application as novel probiotic strain to protect livestock and enhance growth performance.

Properties of Probiotics Kocuria SM1 and Rhodococcus SM2 Isolated from Fish Guts.

This study characterized probiotics Kocuria SM1 and Rhodococcus SM2, which were recovered from the intestinal microbiota of rainbow trout (Oncorhynchus mykiss, Walbaum). The cultures were Gram-positive, non-motile, catalase-positive and oxidase-negative cocci or rods. Cell multiplication of SM1 and SM2 was observed at 4-37°C (45°C for SM1), in 0-20% (w/v) NaCl and at pH2-11. The viability was not affected when exposed to pepsin at pH2.0 and 3.0, and pancreatin at pH8.0. Neither isolates were chrome azurol S-positive for siderophore production. Of the 19 common enzymes analysed using the API-ZYM system, only 8 were evident in the culture of SM1 compared to 11 enzymes for SM2. The secondary metabolites of both probiotics were inhibitory to Acinetobacter baumannii, Vibrio anguillarum and V. ordalii; SM2 inhibited Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa and Staphylococcus aureus. SM2 was resistant to penicillin and sulphatriad, out of six antimicrobial agents; SM1 was resistant to sulphatriad. These results suggest that Kocuria SM1 and Rhodococcus SM2 are able to grow over a wide range of temperature, salinity and pH, including in conditions that mimic the gastrointestinal environment of fish and produce extracellular enzymes that may have a role in the host digestive processes. Importantly, Rhodococcus SM2 displays a high degree of bacteriocinogenic potential against multi-drug-resistant human pathogens that have never been documented among the gut microbiota of fish.

Bacteriocinogenic Potential of Enterococcus faecium Isolated from Wine.

A total of 145 lactic acid bacteria isolated from a variety of Turkish red wines during malolactic fermentation were screened to find bacteriocin-producing strains. Among them, 14 isolates of Enterococcus faecium were identified to produce bacteriocins. PCR screening revealed that some isolates harbored entA and entB genes while some harbored entA, entB and entP genes. An isolate designated as Ent. faecium H46 was selected to characterize its bacteriocins. The bacteriocins were purified to homogeneity from culture supernatant by Amberlite XAD-16, cation-exchange and reverse-phase chromatography. MALDI-TOF mass spectrometry analysis identified the bacteriocins as enterocin A and enterocin B. The presence of Ent. faecium is noteworthy since it is not associated with wine fermentation. However, it has been reported as an important wine spoilage organism due to its potential to produce tyramine. Although species of Enterococcus is not known as wine bacteria, contamination by Ent. faecium may arise from grapes or wineries equipments used for wine production.