Assessment of the safety and anti-inflammatory effects of three Bacillus strains in the respiratory tract.

Chronic respiratory diseases are part of accumulating health problems partly due to worldwide increase in air pollution. By their antimicrobial and immunomodulatory properties, some probiotics constitute promising alternatives for the prevention and treatment of chronic respiratory diseases. We have isolated Bacillus strains from Korean fermented foods and selected three potentially probiotic strains (two Bacillus subtilis and one Bacillus amyloliquefaciens) based on safety, antimicrobial efficacy, activity against airborne pathogens and their immunomodulatory properties in vivo. Safety evaluation included in silico analysis for confirming absence of virulence genes. Safety for the respiratory tract was confirmed by an in vivo pathogenicity test using a murine model. Antimicrobial activity was displayed against several airborne pathogens. Potential antimicrobial metabolites such as 2,3-butanediol and propylene glycol were identified as possible antagonistic agents. Immunomodulatory properties in vitro were confirmed by upregulation of IL-10 expression in a macrophage cell line. Intranasal instillation and inhalation in an ovalbumin (OVA)-induced lung inflammation murine model reduced T helper type 2 (Th2) cytokines at transcriptional and protein levels in the lungs. The safety and potentially beneficial role of these Bacillus strains could be demonstrated for the respiratory tract of a murine model.

Technological properties of beneficial bacteria from the dairy environment and development of a fermented milk with the beneficial strain Lactobacillus casei MRUV6.

In this research paper we describe the technological properties of beneficial lactic acid bacteria (LAB) obtained from a dairy production chain and the development of a fermented milk produced with Lactobacillus casei MRUV6. Fifteen LAB isolates (Lactobacillus sp., Pediococcus sp. and Weissela sp.) presented acidifying abilities (pH ranges from 0.73 to 2.11), were able to produce diacetyl (except by 5 isolates) and exopolysaccharides, and two were proteolytic. L. casei MRUV6 was selected for producing a fermented milk, stored up to 35 d at 4 and 10°C. Counts on MRS agar with added vancomycin (10 mg/l) and MRS agar with added bile salts (1.5% w/v) ranged from 9.7 to 9.9 log CFU/g, independently of the tested conditions, indicating stability and intestinal resistance of L. casei MRUV6, despite some significant differences (P < 0.05). The study demonstrated the technological potential of a potential probiotic candidate strain, L. casei MRUV6, to be used as a starter culture in the dairy industry.

Beneficial properties of lactic acid bacteria naturally present in dairy production.

BACKGROUND: Consumers are increasingly demanding for natural and beneficial foods, in order to improve their health and well-being. Probiotics play an important role in such demand, and dairy foods are commonly used as vehicles for such bacteria, represented predominantly by lactic acid bacteria. Due to consumers demand, food industry is constantly looking for novel bacterial strains, leading to studies that aims the isolation and characterization of their beneficial features. This study aimed to characterize the naturally occurring lactic acid bacteria obtained from a dairy environment, in order to assess their potential use as probiotics. RESULTS: Preliminary screening and PCR analysis, based on 16S rRNA sequencing, were applied to select and identify 15 LAB strains from the genera Lactobacillus (n = 11), Pediococcus (n = 2) and Weissella (n = 2). All strains showed resistance to low pH and the evaluated bile salt concentrations in vitro. The API ZYM test characterized the enzymatic activity of the strains, and a high ß-galactosidase activity was observed in 13 strains. All strains presented resistance to simulated gastric (3 h) and intestinal (4 h) conditions in vitro, the ability to auto- and co-aggregate with indicator microorganisms and a high cell surface hydrophobicity. Most of the strains were positive for map and EFTu beneficial genes. All strains exhibited strong deconjugation of bile salts in vitro and all assimilated lactose. CONCLUSIONS: The phenotypes exhibited in vitro and the presence of beneficial genes revealed the beneficial potential of the studied strains, demanding further analyses in a food matrix and in vivo to allow the development of a functional product, with health-related properties.

Combined effect of bacteriocin produced by Lactobacillus plantarum ST8SH and vancomycin, propolis or EDTA for controlling biofilm development by Listeria monocytogenes.

The Listeria monocytogenes strains selected in the present study exhibited similar behavior in biofilm formation, independently of the tested conditions (bacteriocin from L. plantarum ST8SH, vancomycin, propolis (a natural antimicrobial product) and EDTA (chelating agent)), individual or in associations. The individual application of vancomycin had better inhibitory activity than that of propolis and EDTA; however, the association of the previously mentioned antimicrobial agents with bacteriocins resulted in better performance. However, when we compared the effects of vancomycin, propolis and EDTA, we could clearly observe that the combined application of bacteriocin and vancomycin was more effective than the combination of bacteriocin and propolis, and bacteriocin and EDTA. Considering the current need to reduce the use of antimicrobials and chemical substances in food processing, propolis can represent an alternative to improve the inhibitory effect of bacteriocins against L. monocytogenes biofilm formation, based on the obtained results. In general, high concentrations of bacteriocin produced by L. plantarum ST8SH were more effective in biofilm inhibition, and similar results were observed for vancomycin and propolis; however, all tested EDTA concentrations had similar effect on biofilm formation.

Evaluation of the probiotic potential and effect of encapsulation on survival for Lactobacillus plantarum ST16Pa isolated from papaya.

Capability to produce antilisterial bacteriocins by lactic acid bacteria (LAB) can be explored by the food industry as a tool to increase the safety of foods. Furthermore, probiotic activity of bacteriogenic LAB brings extra advantages to these strains, as they can confer health benefits to the consumer. Beneficial effects depend on the ability of the probiotic strains to maintain viability in the food during shelf-life and to survive the natural defenses of the host and multiply in the gastrointestinal tract (GIT). This study evaluated the probiotic potential of a bacteriocinogenic Lactobacillus plantarum strain (Lb. plantarum ST16Pa) isolated from papaya fruit and studied the effect of encapsulation in alginate on survival in conditions simulating the human GIT. Good growth of Lb. plantarum ST16Pa was recorded in MRS broth with initial pH values between 5.0 and 9.0 and good capability to survive in pH 4.0, 11.0 and 13.0. Lb. plantarum ST16Pa grew well in the presence of oxbile at concentrations ranging from 0.2 to 3.0%. The level of auto-aggregation was 37%, and various degrees of co-aggregation were observed with different strains of Lb. plantarum, Enterococcus spp., Lb. sakei and Listeria, which are important features for probiotic activity. Growth was affected negatively by several medicaments used for human therapy, mainly anti-inflammatory drugs and antibiotics. Adhesion to Caco-2 cells was within the range reported for other probiotic strains, and PCR analysis indicated that the strain harbored the adhesion genes mapA, mub and EF-Tu. Encapsulation in 2, 3 and 4% alginate protected the cells from exposure to 1 or 2% oxbile added to MRS broth. Studies in a model simulating the transit through the GIT indicated that encapsulated cells were protected from the acidic conditions in the stomach but were less resistant when in conditions simulating the duodenum, jejunum, ileum and first section of the colon. To our knowledge, this is the first report on a bacteriocinogenic LAB isolated from papaya that presents application in food biopreservation and may be beneficial to the consumer health due to its potential probiotic characteristics.

Research Trends in the Study of Edible Mushrooms: Nutritional Properties and Health Benefits.

The consumption of edible-culinary mushrooms for the prevention and treatment of chronic disease has gained increasing attention. This review summarizes trends in the biotechnological and medicinal potential of edible mushrooms cultivated worldwide. Macronutrients (fatty acids, amino acids, and carbohydrates), bioactive compounds (phenolics, flavonoids, and carotenoids), and health benefits (antioxidant, antimicrobial, antifungal, anticancer, and pre-biotics properties) of mushrooms are described, including their cultivation, industrial processing, and consumption. In general, edible-culinary mushrooms present a rich nutritional composition with beneficial properties for human health. Indeed, the consumption of edible mushrooms is associated with a reduction in the risk of cancer and diabetes. Furthermore, mushrooms can be incorporated into different food formulations and used as a medicinal substance due to their mycochemicals with antioxidant capacity. Edible mushrooms are considered a "superfood" and can be recommended as a valuable constituent in the daily diet. In conclusion, this review describes trends, future decision-making, and guidance on the health benefits of edible mushrooms.

Characterization of a bacteriocin produced by Lactobacillus sakei R1333 isolated from smoked salmon.

Strain R1333, isolated from commercially available smoked salmon, was identified as Lactobacillus sakei based on biochemical tests, sugar fermentation reactions (API 50 CHL), PCR with species-specific primers and sequencing of the 16S rRNA gene. Strain R1333 produces a 3811 kDa class IIa bacteriocin, active against Streptococcus caprinus, Streptococcus macedonicus, Streptococcus spp., L. sakei, Lactococcus lactis subsp. lactis, Listeria innocua, Listeria ivanovii subsp. ivanovii and Listeria monocytogenes. The mode of activity against L. innocua 2030C and L. ivanovii subsp. ivanovii ATCC 19119 was bactericidal, resulting in cell lysis and enzyme- and DNA-leakage. The highest level of activity (1600 AU/mL) was recorded when cells were grown at 30°C in MRS broth (initial pH 6.5). Only 800 AU/mL was recorded when strain R1333 was grown in MRS without Tween 80. Lower levels of bacteriocin production were recorded when strain R1333 was grown in MRS at 20°C. Peptide R1333 adsorbs at low levels (200 AU/mL) to producer cells. Purification of bacteriocin R1333 was performed by 60% ammonium sulfate precipitation, followed by separation on a SepPak C(18) column and reverse-phase HPLC on a Nucleosil C(18) column with a linear gradient from 0.1% TFA to 90% acetonitryl. A molecular mass of 3811 kDa was determined by mass spectrometry. Based on mass spectrometry and sequencing of the PCR amplified fragment targeting the sakG gene, L. sakei R1333 is a potential producer of sakacin G. This is the first report of the identification of sakacin G produced by L. sakei isolated from smoked salmon.

Formation of Free Amino Acids and Bioactive Peptides During the Ripening of Bulgarian White Brined Cheeses.

Bioactive peptides and free amino acids obtained from Bulgarian goat, sheep and cow white brined cheeses, produced with same starter culture, during ripening were evaluated. The concentration of total free amino acids was increasing in all tested cheeses in the first 30 days of ripening. In the next 30 days in sheep cheeses, the concentration increased as recorded for most of the amino acids. Amino acids with highest levels detected throughout the whole ripening period in goat, sheep and cow cheese types were leucine, phenylalanine, arginine, valine and lysine. MALDI-TOF analysis of evaluated cheeses resulted in detection of production of bioactive peptide derivates from milk proteins: 51 peptides in cow, 31 peptides in sheep and 22 peptides in goat cheeses. Peptide αs1-CN (f35-40) was found only in cow cheese. In cow cheese, higher intensity was detected for αs1-CN (f1-9) and ß-CN (f194-203 and f203-219) peptides. In goat cheese was recorded αs1-CN peptides, and there was a tendency to increase the peptides released from ß-CN, with the highest intensity of fragments αs1-CN (f1-9 and f24-30) and ß-CN (f194-209 and f203-219). In sheep cheese, the recorded primarily peptides were αs1-CN and peptides released from ß-CN. Different bioactive peptides, derivate from casein, were detected as follows: 6 peptides were ACE inhibitory peptides, 3 peptides were αS1-casokinins, 1 peptide was caseinophopeptide, 1 peptide was immunopeptide. Twelve bioactive peptides were recorded to be derivates from ß-casein: 1 peptide was ACE peptide, 4 peptides were caseino-phosphopeptides, 1 peptide was immunopeptide, 1 peptide ß-casokinin, 1 antibacterial peptide and 4 multifunctional peptides. Of peptides released by proteolysis of αS2-CN was found 1 bioactive peptide with antimicrobial activity. On our best knowledge, this paper contributes new data about free amino acids and bioactive peptides in the connection between type of milk and period for cheese ripening in the Bulgarian goat, sheep and cow white brined cheeses.

Safety Evaluation and In vivo Strain-Specific Functionality of Bacillus Strains Isolated from Korean Traditional Fermented Foods.

Unveiling and understanding differences in physiological features below the species level may serve as an essential fast-screening tool for selecting strains that can promote a specific probiotic effect. To study the intra-species diversity of Bacillus, a genus with a wide range of enzyme activities and specificity, 190 Bacillus strains were isolated from traditional Korean fermented food products. Altogether, in the preliminary safety screening, 8 of these strains were found negative for lecithinase and hemolysis activity and were selected for further investigations. On the basis of different levels of enzyme functionalities (high or low proteolytic, amylolytic, and lipolytic (PAL) activities), two Bacillus subtilis strains were selected for an in vivo study. Each of the two strains was separately administered at a level of 1 × 108 CFU per day to C57BL/6 mice that were fed 60% high-fat diet ad libitum for 8 weeks, while Xenical, an anti-obesity drug, was used as a positive control in the experimental setup. B. subtilis M34 and B. subtilis GS40a with low and high amylolytic activities, respectively, induced significantly different and contrasting physiological effects. The production of short-chain fatty acids appeared to be closely associated with a shift in the gut microbiota.

Bacteriocins - exploring alternatives to antibiotics in mastitis treatment.

Mastitis is considered to be the most costly disease affecting the dairy industry. Management strategies involve the extensive use of antibiotics to treat and prevent this disease. Prophylactic dosages of antibiotics used in mastitis control programmes could select for strains with resistance to antibiotics. In addition, a strong drive towards reducing antibiotic residues in animal food products has lead to research in finding alternative antimicrobial agents. In this review we have focus on the pathogenesis of the mastitis in dairy cows, existing antibiotic treatments and possible alternative for application of bacteriocins from lactic acid bacteria in the treatment and prevention of this disease.

Mode of action and In Vitro susceptibility of mastitis pathogens to macedocin ST91KM and preparation of a teat seal containing the bacteriocin.

Mastitis is considered to be the most economically costly disease affecting the dairy industry. Regular dosage of animals with antibiotics, including use of prophylactic concentrations, may select for resistant strains. The purpose of this study was to determine the mode of action of a new bacteriocin (macedocin ST91KM), to evaluate the antimicrobial resistance of mastitis pathogens to antibiotics commonly used in treatment remedies, and to introduce the possible use of an alternative antimicrobial agent. The bacteriocin macedocin ST91KM, produced by Streptococcus gallolyticus subsp. macedonicus ST91KM, is bactericidal to Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus uberis and Staphylococcus aureus associated with mastitis infections, including strains resistant to methicillin and oxacillin. Sensitive cells were deformed and secreted nucleotides, K(+) and ß-galactosidase when exposed to macedocin ST91KM. Adsorption of the peptide to target cells decreased in the presence of solvents, suggesting that receptors on the cell surfaces have lipid moieties. No adsorption was recorded in the presence of MgCl2, KI and Na2CO3, suggesting that ionic strength plays an important role. A teat seal preparation containing macedocin ST91KM effectively released the peptide and inhibited the growth of S. agalactiae. Macedocin ST91KM could form the basis for alternative dry cow therapy to prevent mastitis infections in dairy cows as it is effective against pathogens that display resistance to conventional antibiotic therapy.

Bacteriocins From LAB and Other Alternative Approaches for the Control of Clostridium and Clostridiodes Related Gastrointestinal Colitis.

The gut microbiome is considered as a promising target for future non-conventional therapeutic treatment of inflammatory and infectious diseases. The search for appropriate safe and beneficial (lactic acid bacterial and other) putative probiotic strains and/or their antimicrobial metabolites represents a challenging approach for combating several problematic and emerging infections. The process of selecting suitable strains, especially of lactic acid bacteria (LAB) with superior properties, has been accelerated and intensified during the past two decades, also thanks to recent developments in lab techniques. Currently, special focus is on the potential of antimicrobial metabolites produced by some LAB strains and their application as active therapeutic agents. The vision is to develop a scientific basis for 'biotherapeutics' as alternative to conventional approaches in both human and veterinary medicine. Consequently, innovative and promising applications of LAB to the therapeutic practice are presently emerging. An overview of the existing literature indicates that some antimicrobial metabolites such as bacteriocins, widely produced by different bacterial species including LAB, are promising biotherapeutic agents for controlling infections caused by potential pathogens, such as Clostridium and Clostridiodes. Non-conventional, safe and well designed therapeutic treatments may contribute to the improvement of gut dysbiotic conditions. Thereby gut homeostasis can be restored and inflammatory conditions such as gastrointestinal colitis ameliorated. Combining the knowledge on the production, characterization and application of bacteriocins from probiotic LAB, together with their antibacterial properties, appears to be a promising and novel approach in biotherapy. In this overview, different scenarios for the control of Clostridium spp. by application of bacteriocins as therapeutic agents, also in synergistic combination with antibiotics, will be discussed.

Effect of modified MRS medium on production and purification of antimicrobial peptide ST4SA produced by Enterococcus mundtii.

Highest antimicrobial activity of peptide ST4SA (51,200 AU/mL) was recorded after 14 h of growth in MRS broth with optimal production at pH 6.0 or 6.5. Growth of strain ST4SA in the presence of tryptone, yeast extract, or a combination of the two, yielded 102,400 AU/mL. An increase in production of peptide ST4SA to 102,400 AU/mL was recorded in the presence of 20.0 g/L fructose, but decreased to 25,600 AU/mL in the presence of lactose (20.0 g/L) or mannose (20.0 g/L) as sole carbon source. Lower activity (25,600 AU/mL) was recorded when 2.0 g/L K(2)HPO(4) was replaced by 2.0 g/L KH(2)PO(4) in MRS broth. An increase of K(2)HPO(4) to 10.0 g/L and 20.0g/L resulted in higher activity (102,400 AU/mL). Addition of glycerol to MRS broth had a negative effect on peptide ST4SA production. Production of peptide ST4SA required the presence of magnesium sulphate, manganese sulphate and 5.0 g/L sodium acetate. Exclusion of tri-ammonium citrate from the medium resulted in reduction of activity to 3,200 AU/mL. Maximum activity (102,400 AU/mL) was recorded in MRS supplemented with 1.0 ppm Vit. C, DL-6,8-thioctic acid or thiamine, respectively. Growth of Listeria ivanovii susbp. ivanovii ATCC 19119 in the presence of peptide ST4SA (12,800 AU/mL) resulted in 99% cell lysis after 18 h. Improved production of peptide ST4SA was recorded in MRS broth (Biolab) pre-treated with Amberlite XAD-1180. Precipitation with ammonium sulphate, followed by gel filtration chromatography, yielded the highest level of peptide ST4SA. This paper describes the partially deproteination of growth medium to facilitate peptide ST4SA purification.

Phenotypic and genetic heterogeneity of lactic acid bacteria isolated from "Alheira", a traditional fermented sausage produced in Portugal.

The aim of this study was to evaluate the phenotypic and genetic heterogeneity of lactic acid bacteria (LAB) isolated from "Alheira", a fermented sausage produced in Portugal. LAB were identified to genus and species level by phenotypic characteristics, using genus or species-specific primers and sequencing of the gene encoding 16S rRNA. Two-hundred and eighty-three isolates were grouped into 14 species. Lactobacillus plantarum was isolated from all sausages and Enterococcusfaecalis from most of the samples. Low numbers of Lactobacillus paraplantarum, Lactobacillus brevis, Lactobacillus rhamnosus, Lactobacillus sakei, Lactobacillus zeae, Lactobacillus paracasei, Leuconostoc mesenteroides, Pediococcus pentosaceus, Pediococcus acidilactici, Weissella cibaria, Weissella viridescens and Enterococcus faecium were recorded. The genetic heterogeneity of L. plantarum and E. faecalis strains were determined by numerical analysis of DNA banding patterns obtained by RAPD-PCR. Strains of L. plantarum and E. faecalis were different from different producers. This study forms the basis from which starter cultures could be selected for production of "Alheira".

Bacteriocins from Lactobacillus plantarum - production, genetic organization and mode of action: produção, organização genética e modo de ação.

Bacteriocins are biologically active proteins or protein complexes that display a bactericidal mode of action towards usually closely related species. Numerous strains of bacteriocin producing Lactobacillus plantarum have been isolated in the last two decades from different ecological niches including meat, fish, fruits, vegetables, and milk and cereal products. Several of these plantaricins have been characterized and the aminoacid sequence determined. Different aspects of the mode of action, fermentation optimization and genetic organization of the bacteriocin operon have been studied. However, numerous of bacteriocins produced by different Lactobacillus plantarum strains have not been fully characterized. In this article, a brief overview of the classification, genetics, characterization, including mode of action and production optimization for bacteriocins from Lactic Acid Bacteria in general, and where appropriate, with focus on bacteriocins produced by Lactobacillus plantarum, is presented.

Bacteriocin production by Lactobacillus plantarum AMA-K isolated from Amasi, a Zimbabwean fermented milk product and study of the adsorption of bacteriocin AMA-K TO Listeria sp.

Bacteriocin AMA-K produced by Lactobacillus plantarum AMA-K inhibits the growth of Enterococcus spp., Escherichia coli, Klebsiella pneumoniae and Listeria spp. Growth of strain AMA-K in BHI, M17, soy milk and molasses was similar to growth in MRS. The effect of organic nitrogen sources, carbohydrates, glycerol, K2HPO4 and KH2PO4, MgSO4, MnSO4, tri-ammonium citrate, Tween 80, vitamins and initial pH on bacteriocin AMA-K was determined. The mode of action of bacteriocin AMA-K was studied. The effect of bacteriocin AMA-K to actively growing Listeria innocua LMG13568, L. ivanovii subsp. ivanovii ATCC19119 and L. monocytogenes ScottA was determined. Adsorption of bacteriocin AMA-K to target cells at different temperatures, pH and in presence of Tween 20, Tween 80, ascorbic acid, potassium sorbate, sodium nitrate and sodium chloride were studied. Bacteriocin AMA-K shares high homology to pediocin PA-1.

Lactic Acid Bacteria (LAB) and Their Bacteriocins as Alternative Biotechnological Tools to Control Listeria monocytogenes Biofilms in Food Processing Facilities.

Bacteriocins are antimicrobial peptides produced by bacteria Gram-negative and Gram-positive, including lactic acid bacteria (LAB), organisms that are traditionally used in food preservation practices. Bacteriocins have been shown to have an aptitude as biofilm controlling agents in Listeria monocytogenes biofilms, a major risk for consumers and the food industry. Biofilms protect pathogens from sanitization procedures, allowing them to survive and persist in processing facilities, resulting in the cross-contamination of the end products. Studies have been undertaken on bacteriocinogenic LAB, their bacteriocins, and bioengineered bacteriocin derivatives for controlling L. monocytogenes biofilms on different surfaces through inhibition, competition, exclusion, and displacement. These alternative strategies can be considered promising in preventing the development of resistance to conventional sanitizers and disinfectants. Bacteriocins are "friendly" antimicrobial agents, and with high prevalence in nature, they do not have any known associated public health risk. Most trials have been carried out in vitro, on food contact materials such as polystyrene and stainless steel, while there have been few studies performed in situ to consolidate the results observed in vitro. There are strategies that can be employed for prevention and eradication of L. monocytogenes biofilms (such as the establishment of standard cleaning procedures using the available agents at proper concentrations). However, commercial cocktails using alternatives compounds recognized as safe and environmental friendly can be an alternative approach to be applied by the industries in the future.

Characterization of two bacteriocins produced by Pediococcus acidilactici isolated from "Alheira", a fermented sausage traditionally produced in Portugal.

Lactic acid bacteria were isolated from "Alheira" sausages that have been sampled from different regions in Portugal. The sausages were produced according to different recipes and with traditional starter cultures. Two isolates (HA-6111-2 and HA-5692-3) from different sausages were identified as strains of Pediococcus acidilactici. Each strain produces a bacteriocin, designated as bacHA-6111-2 and bacHA-5692-3. Both bacteriocins are produced at low levels after 18 h of growth in MRS broth (3200 AU/ml against Enterococcus faecium HKLHS and 1600 AU/ml against Listeria innocua N27). BacHA-6111-2 and bacHA-5692-3 are between 3.5 kDa and 6.5 kDa in size, as determined by tricine-SDS-PAGE. Complete inactivation or significant reduction in antimicrobial activity was observed after treatment of cell-free supernatants with proteinase K, pronase and trypsin. No change in activity was recorded when treated with catalase. Both bacteriocins are sensitive to treatment with Triton X-114 and Triton X-100, but resistant to Tween 20, Tween 80, SDS, Oxbile, NaCl, urea and EDTA. The bacteriocins remained stable after 2 h at pH 6.0. A decrease in antibacterial activity was recorded after 60 min at 100 degrees C. After 60 min at 80 degrees C, 60 degrees C and 25 degrees C the antibacterial activity against L. innocua N27 decreased by 25%. Addition of bacHA-6111-2 and bacHA-5692-3 (1600 AU/ml) to a mid-log (5-h-old) culture of L. innocua N27 inhibited growth for 7 h. Addition of the bacteriocins (3200 AU/ml) to a mid-log (5-h-old) culture of E. faecium HKLHS repressed cell growth. The bacteriocins did not adhere to the surface of the producer cells. Both strains contain a 1044 bp DNA fragment corresponding in size to that recorded for pediocin PA-1. Sequencing of the fragments from both bacteriocins revealed homology to large sections of pedA (188 bp), pedB (338 bp) and pedC (524 bp) of pediocin PA-1 and the bacteriocins are considered similar to pediocin PA-1.

An antibacterial and antiviral peptide produced by Enterococcus mundtii ST4V isolated from soya beans.

Enterococcus mundtii ST4V, isolated from soya beans, produces a 3950Da antibacterial peptide active against Gram-positive and Gram-negative bacteria, including Enterococcus faecalis, Streptococcus spp., Pseudomonas aeruginosa, Klebsiella pneumoniae, Streptococcus pneumoniae and Staphylococcus aureus. The peptide also inactivated the herpes simplex viruses HSV-1 (strain F) and HSV-2 (strain G), a polio virus (PV3, strain Sabin) and a measles virus (strain MV/BRAZIL/001/91, an attenuated strain of MV). MV, HSV-1 and HSV-2 were 95.5%-99.9% inactivated by peptide ST4V at 400 microg/ml. Monkey kidney Vero cells were not inactivated, even at four times the level peptide ST4V displayed antiviral activity, indicating that the effect was not due to cytotoxicity. Complete inactivation or significant reduction in antimicrobial activity was observed after treatment of peptide ST4V with Proteinase K, pronase, pepsin and trypsin. No change in antimicrobial activity was recorded after treatment with alpha-amylase, suggesting that peptide ST4V was not glycosylated. This is the first description of an antibacterial and antiviral peptide with such broad-spectrum of activity, produced by a lactic acid bacterium.

Optimization of bacteriocin ST311LD production by Enterococcus faecium ST311LD, isolated from spoiled black olives.

Bacteriocin ST311LD is approximately 2.3 kDa in size. Low levels of bacteriocin activity were recorded in BHI and M17 broth (800 AU/ml) and in 10% (w/v) soy milk (3,200 AU/ml). No bacteriocin production was recorded in 10% (w/v) molasses, despite good growth. Optimal levels (12,800 AU/ml) were detected in MRS broth which had been supplemented with tryptone (20.0 g/l), saccharose (5.0 or 10.0 g/l) or vitamin C (1 ppm). Increased potassium levels did not result in higher levels of activity, and glycerol (1.0 g/l) inhibited the production of bacteriocin ST311LD.