Bacteriocin-producing Lactic Acid Bacteria Strains Research Articles

Bacteriocin-Producing Lactic Acid Bacteria Strains with Antimicrobial Activity Screened from Bamei Pig Feces.

Lactic acid bacteria (LAB), which are characterized by producing various functional metabolites, including antioxidants, organic acids, and antimicrobial compounds, are widely used in the food industry to improve gut health and prevent the growth of spoilage microorganisms. With the continual incidence of foodborne disease and advocacy of consumers for gut health, LAB have been designated as vital biopreservative agents in recent years. Therefore, LAB with excellent antimicrobial properties and environmental tolerance should be explored further. In this study, we focus on screening the LAB strains from a specialty pig (Bamei pig) feces of the Tibetan plateau region and determine their antimicrobial properties and environmental tolerance to evaluate their potential probiotic values. A total of 116 LAB strains were isolated, from which the LAB strain Qinghai (QP)28-1 was identified as Lactiplantibacillus (L.) plantarum subsp. plantarum using 16S rDNA sequencing and recA amplification, showing the best growth capacity, acid production capacities, environmental tolerance, hydrophobicity, antibiotic susceptibility, and bacteriocin production capacity. Furthermore, this strain inhibited the growth of multiple pathogens by producing organic acids and bacteriocin. These bacteriocin-encoding genes were identified using PCR amplification, including plnS, plnN, and plnW. In conclusion, bacteriocin-producing L. plantarum subsp. plantarum QP28-1 stands out among these 116 LAB strains, and was considered to be a promising strain used for LAB-related food fermentation. Moreover, this study provides a convenient, comprehensive, and shareable profile for screening of superior functional and bacteriocin-producing LAB strains, which can be used in the food industry.

Evaluation of bacteriocin-producing lactic acid bacteria strains isolated from “nem chua Thu Duc”

This study aimed to select some lactic acid bacteria (LAB) strains to produce bacteriocin isolated from two different local brand names of nem chua Thu Duc, a Vietnamese traditional fermented meat product. Eight different LAB strains were isolated from products. However, only Pediococcus pentosaceus isolated from 72-h fermented “Ba Chin” products and Weissella paramesenteroides isolated from 120-h fermented “Diem” products were selectively screened for their bacteriocin production. The antimicrobial activity was detected by the agar diffusion method against Escherichia coli ATCC 25922, Salmonella enterica serovar Typhimurium ATCC 13076, Staphylococcus aureus ATCC 6538, and Bacillus cereus ATCC 25924 used as target strains. Bacteriocin of Pediococcus pentosaceus strain showed a high antibacterial ability with a sterile ring diameter that produces an average of 12.20 - 16.07 mm. Bacteriocin of Weissella paramesenteroides strain only showed inhibitory activity on the growth of indicator microorganisms with an average inhibitory ring diameter of 9.75 - 15.27 mm. In conclusion, it seems like both bacteriocin-producing P. pentosaceus and W. paramesenteroides give potential applications as a starter culture, bio-preservation and bio-safety control in fermented meat as well as other kinds of fermented foods.

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.

Application of Bacteriocins in Meat and Meat Products: An Update

: Being an important source of human enteric diseases, microbiological safety is one of the major risk concerns in the meat industry. In order to inhibit and inactivate microbial contamination and extend the shelf life of meat products, different procedures have been practiced, including the addition of bacteriocins as proteinaceous antagonistic preservatives. This article discusses the application of bacteriocins which are capable of controlling the growth of pathogenic and spoilage microorganisms in meat and meat products. We identify possible ways to improve the performance of bacteriocins ensuring food safety and toxicity. We first provide a brief introduction to the classification of bacteriocins and then discuss their antimicrobial properties and mechanism of action alone and in combination with other hurdles in meat and meat products. Moreover, application methods of bacteriocins in meat products are described and cross-compared, introducing emerging meat products containing bacteriocins. : Despite the existence of many reports related to the application of bacteriocin-producing strains of lactic acid bacteria in meat products, very few review articles have attempted at evaluating the application of bacteriocins in the red meat while observing their antimicrobial mechanism of action as well as evaluating their applications in meat products. The application of these proteins in meat products has received considerable attention; however, there are still some drawbacks and limitations for their application. Characterization, identification, toxicity evaluation and investigating application level of bacteriocins produced by meat borne/non-meat borne bacteria appears to be necessary in order to increase the efficiency of extending shelf life and improving the microbial stability of meat products.

The Potential of Class II Bacteriocins to Modify Gut Microbiota to Improve Host Health.

Production of bacteriocins is a potential probiotic feature of many lactic acid bacteria (LAB) as it can help prevent the growth of pathogens in gut environments. However, knowledge on bacteriocin producers in situ and their function in the gut of healthy animals is still limited. In this study, we investigated five bacteriocin-producing strains of LAB and their isogenic non-producing mutants for probiotic values. The LAB bacteriocins, sakacin A (SakA), pediocin PA-1 (PedPA-1), enterocins P, Q and L50 (enterocins), plantaricins EF and JK (plantaricins) and garvicin ML (GarML), are all class II bacteriocins, but they differ greatly from each other in terms of inhibition spectrum and physicochemical properties. The strains were supplemented to mice through drinking water and changes on the gut microbiota composition were interpreted using 16S rRNA gene analysis. In general, we observed that overall structure of the gut microbiota remained largely unaffected by the treatments. However, at lower taxonomic levels, some transient but advantageous changes were observed. Some potentially problematic bacteria were inhibited (e.g., Staphylococcus by enterocins, Enterococcaceae by GarML, and Clostridium by plantaricins) and the proportion of LAB was increased in the presence of SakA-, plantaricins- and GarML-producing bacteria. Moreover, the treatment with GarML-producing bacteria co-occurred with decreased triglyceride levels in the host mice. Taken together, our results indicate that several of these bacteriocin producers have potential probiotic properties at diverse levels as they promote favorable changes in the host without major disturbance in gut microbiota, which is important for normal gut functioning.

Protection of honeybee Apis mellifera by its endogenous and exogenous lactic flora against bacterial infections

Abstract Lactic acid bacteria (LAB) symbionts of honeybees are certainly playing key roles in their host's colony functioning. The defense against bacterial pathogen invasion by endogenous LAB has been considered as promising and usable phenomenon. This study addresses bacteriocinogeny as one of antibacterial action mode displayed by bacteria. The honeybee endogenous LAB isolated from worker honeybee intestines (61 strains), queen honeybee intestines (16 strains) and beebread (25 strains) were tested for bacteriocin production ability. We checked also well characterized bacteriocin producing LAB strains against bacteria causing American foulbrood (AFB) – Paenibacillus larvae aiming possible use of exogenous LAB for control AFB in honeybees and in the same time to observe the vulnerability of endogenous bacteria exposed to bacteriocin producers. We demonstrated that none of 102 studied LAB strains, isolated from worker honeybee intestines, from queen honeybee intestines and from beebread, produced bacteriocins detectable by the well diffusion method (WDM). All of them failed to inhibit two strains of P. larvae . Three exogenous bacteriocin-producing LAB strains were tested against the same pathogens and against 25 endogenous bacterial isolates representing 11 different LAB species. The screening showed that all the tested exogenous bacteriocin-producing strains inhibited the tested P. larvae strains. The endogenous LAB strains exhibited varied sensitivity profiles when treated with bacteriocin-producing strains. This raises similar challenges to those observed in antibiotic applications leading to dysbacteriosis, even though the efficacy of these bacteriocins against P. larvae in an in vitro system is evident.

Quantitative Appraisal of the Probiotic Attributes and In Vitro Adhesion Potential of Anti-listerial Bacteriocin-producing Lactic Acid Bacteria.

Estimation of bile tolerance, endurance to gastric and intestinal environment and adhesion potential to intestinal cells are significant selection criteria for probiotic lactic acid bacteria (LAB). In this paper, the probiotic potential of native bacteriocin-producing LAB isolated previously from indigenous source has been determined through quantitative approaches. Among fifteen anti-listerial bacteriocin-producing native LAB, ten strains were found to be bile tolerant. The presence of bile salt hydrolase (bsh) gene in native Lactobacillus plantarum strains was detected by PCR and confirmed by nucleic acid sequencing of a representative amplicon. Interestingly, three native LAB strains exhibited significant viability in simulated gastric fluid, analogous to the standard LAB Lactobacillus rhamnosus GG, while an overwhelming majority of the native LAB strains demonstrated the ability to survive and remain viable in simulated intestinal fluid. Quantitative adhesion assays based on conventional plating method and a fluorescence-based method revealed that the LAB isolates obtained from dried fish displayed significant in vitro adhesion potential to human adenocarcinoma HT-29 cells, and the adhesion level was comparable to some of the standard probiotic LAB strains. The present study unravels putative probiotic attributes in certain bacteriocin-producing LAB strains of non-human origin, which on further in vivo characterization could find specific applications in probiotic food formulations targeted for health benefits.

Intracellular pH of Mycobacterium avium subsp. paratuberculosis following exposure to antimicrobial compounds monitored at the single cell level

Mycobacterium avium subsp. paratuberculosis (MAP) is the etiologic agent of Johne's disease; moreover, it seems to be implicated in the development of Crohn's disease in humans. In the present study, fluorescence ratio imaging microscopy (FRIM) was used to assess changes in intracellular pH (pH i) of one strain of MAP after exposure to nisin and neutralized cell-free supernatants (NCSs) from five bacteriocin-producing lactic acid bacteria (LAB) with known probiotic properties. The evaluation of pH i by FRIM provides information about the physiological state of bacterial cells, bypassing the long and problematic incubations needed for methods relying upon growth of MAP such as determination of colony forming units. The FRIM results showed that both nisin and the cell-free supernatant from Lactobacillus plantarum PCA 236 affected the pH i of MAP within a few hours. However, monitoring the population for 24 h revealed the presence of a subpopulation of cells probably resistant to the antimicrobial compounds tested. Use of nisin and bacteriocin-producing LAB strains could lead to new intervention strategies for the control of MAP based on in vivo application of probiotic cultures as feed additives at farm level.

Changes in the microbial composition of raw milk induced by thermization treatments applied prior to traditional Greek hard cheese processing.

The microbiological quality, safety, and composition of mixtures of ewe's and goat's milk (90:10) used for cheesemaking were evaluated before and after thermization at 60 and 67 degrees C for 30 s. Such mild thermal treatments are commonly applied to reduce natural contaminants of raw milk before processing for traditional hard Greek cheeses. Raw milk samples had an average total bacterial count of 7.3 log CFU/ml; most of these bacteria were lactic acid bacteria (LAB) and pseudomonads. The LAB flora of raw milk was dominated by enterococci (40.8%), followed by lactococci (20.4%), leuconostocs (18.4%), and mesophilic lactobacilli (10.2%). Enterococcus faecalis (30.1%) and Enterococcus faecium (13.7%) were the most common LAB isolates, followed by Enterococcus durans, Lactococcus lactis subsp. lactis, Lactobacillus plantarum, and Leuconostoc lactis. Thermization at 60 degrees C for 30 s was effective for reducing raw milk contamination by enterobacteria (5.1 log CFU/ml), coagulase-positive staphylococci (3.3 log CFU/ml), and Listeria (present in 25-ml samples) to safe levels, but it also reduced mesophilic lactococci, leuconostocs, lactobacilli, and selected enterococci (72.0%) in thermized milk. Thermization at 67 degrees C for 30 s had a major inactivation effect on all bacterial groups. Two nisin-producing L. lactis subsp. lactis strains (M78 and M104) were isolated from raw milk, but neither nisin-producing nor other bacteriocin-producing LAB strains were isolated from thermized milk. Thus, thermization treatments control harmful bacteria but also may have a negative impact on milk quality by reducing desirable LAB and the biodiversity of raw milk bacteria overall, inactivating potentially protective LAB strains and enhancing the ability of potentially pathogenic enterococci to grow in fresh cheese curds.

Preliminary characterization of bacteriocins from Lactococcus lactis, Enterococcus faecium and Enterococcus mundtii strains isolated from turbot ( Psetta maxima)

The aim of this work was the characterization of new strains of lactic acid bacteria (LAB) from farmed fish and with potential application as biopreservatives against both Listeria monocytogenes and Staphylococcus aureus. Twenty-five strains of LAB isolated from the muscle of farmed turbot were investigated. Genetic identification of the bacteriocin-producing LAB strains was performed by means of a PCR method using novel BAL1/BAL2 16S ribosomal-RNA-targeted primers. Maximum bacteriocin production by Lactococcus lactis ssp. lactis USC-39, Enterococcus faecium USC-46 and Enterococcus mundtii USC-51 was detected in the stationary phase of growth. Both acidification and the production of hydrogen peroxide by LAB were ruled out as the source of the inhibition. In contrast, the antimicrobial activity of all three LAB strains was inactivated by the addition of proteinase K, thus confirming the proteinaceous nature of the inhibition. The activity against L. monocytogenes was maintained in the 3.5–5.5 or 3.5–6.5 pH range, depending on the LAB strain. Likewise, inhibition of S. aureus strains was observed in the 3.5–4.5 and in the 3.5–5.5 pH ranges, depending on the LAB strain and on the S. aureus strain tested. Bacteriocin activity was stable in all three strains after heating the cell-free extract for 60 min at 100 °C, or even for 15 min at 121 °C, in all the three LAB strains. The acidic and heat-resistant bacteriocins produced by the three LAB strains isolated from turbot, able to inhibit the growth of both L. monocytogenes and S. aureus may find application as biopreservatives in fermented and/or heated food products.

FUNDAMENTALS AND PERSPECTIVES FOR THE USE OF BACTERIOCINS PRODUCED BY LACTIC ACID BACTERIA IN MEAT PRODUCTS

Bacteriocins of lactic acid bacteria (LAB) are proteinaceous compounds that may present antimicrobial activity towards important foodborne pathogens and spoilage-related microflora. Due to these properties, bacteriocin-producing strains or purified bacteriocins have a great potential of use in biologically based food preservation systems. Despite the growing number of articles describing the isolation of bacteriocinogenic strains, genetic determinants for production, as well as the purification and biochemical characterization of these inhibitory substances, there are only limited reports of successful application of bacteriocins to meats. This paper presents a critical review of the methods available for screening of bacteriocin-producing LAB strains from meats and also discusses the proposed mechanisms of action for LAB bacteriocins. Additionally, an overview of the Brazilian experience in the application of LAB bacteriocins to meats and meat products is given.

Review: Bacteriocins of Lactic Acid Bacteria

During the last few years, a large number of new bacteriocins produced by lactic acid bacteria (LAB) have been identified and characterized. LAB-bacteriocins comprise a heterogeneous group of physicochemically diverse ribosomally-synthesized peptides or proteins showing a narrow or broad antimicrobial activity spectrum against Gram-positive bacteria. Bacteriocins are classified into separate groups such as the lantibiotics (Class I); the small (<10 kDa) heat-stable postranslationally unmodified non-lantibiotics (Class II), further subdivided in the pediocin-like and anti Listeria bacteriocins (subclass IIa), the two-peptide bacteriocins (subclass IIb), and the sec-dependent bacteriocins (subclass IIc); and the large (>30 kDa) heat-labile non-lantibiotics (Class III). Most bacteriocins characterized to date belong to Class II and are synthesized as precursor peptides (preprobacteriocins) containing an N-terminal double-glycine leader peptide, which is cleaved off concomitantly with externalization of biologically active bacteriocins by a dedicated ABC-transporter and its accessory protein. However, the recently identified sec-dependent bacteriocins contain an N-terminal signal peptide that directs bacteriocin secretion through the general secretory pathway (GSP). Most LAB-bacteriocins act on sensitive cells by destabilization and permeabilization of the cytoplasmic membrane through the formation of transitory poration complexes or ionic channels that cause the reduction or dissipation of the proton motive force (PMF). Bacteriocin producing LAB strains protect themselves against the toxicity of their own bacteriocins by the expression of a specific immunity protein which is generally encoded in the bacteriocin operon. Bacteriocin production in LAB is frequently regulated by a three-component signal transduction system consisting of an induction factor (IF), and histidine protein kinase (HPK) and a response regulator (RR). This paper presents an updated review on the general knowledge about physicochemical properties, molecular mode of action, biosynthesis, regulation and genetics of LAB-bacteriocins.

Control of Listeria monocytogenes by bacteriocins and monitoring of bacteriocin-producing lactic acid bacteria by colony hybridization in semi-hard raw milk cheese.

The ability of Listeria monocytogenes to grow at low temperatures and increase to hazardous levels during storage of dairy products is of great concern to the dairy industry. List. monocytogenes was detected in 3·6% of 774 cows' milk samples from 114 farms in Central Spain (Gaya et al. 1998). Ewes' and goats' milks produced in Spain had contamination levels of 2·2 and 2·6% respectively (Rodríguez et al. 1994; Gaya et al. 1996). List. monocytogenes may survive during the manufacture and ripening of most cheese varieties, with higher risk in soft surface-ripened cheeses owing to the rise in pH during ripening (Maisnier-Patin et al. 1992; Giraffa & Carminati, 1997).The efficacy with which actively growing bacteriocin-producing cultures inhibit List. monocytogenes in some cheese varieties has been demonstrated. Nisin-producing starters inhibited List. monocytogenes V7 in Camembert cheese (Maisnier-Patin et al. 1992) and List. innocua BL86/26 in raw ewes' milk Manchego cheese (Rodríguez et al. 1998). A lacticin 3147-producing starter culture (McAuliffe et al. 1999) reduced List. monocytogenes Scott A levels in cottage cheese. Inoculation of milk with enterocin AS-48-producing Enterococcus faecalis INIA 4 inhibited List. monocytogenes strains Ohio and Scott A during the manufacture and ripening of Manchego cheese (Nuñez et al. 1997).Semi-hard cheese varieties from raw or pasteurized cows' milk mixed with different proportions of ewes' and/or goats' milk represent ∼ 50% of Spanish cheese production. The objective of the present work was to evaluate the antilisterial activity of three bacteriocin-producing lactic acid bacteria strains isolated from milk (Rodríguez et al. 2000) during the manufacture and ripening of a semi-hard cheese manufactured from raw milk.