Bacteriocins

Although several strains ofB. subtiliswith antifungal activity have been isolated worldwide, to date there are no published reports regarding the isolation of a nativeB. subtilisstrain from strawberry plants in Mexico. A native bacterium (Bacillus subtilis21) demonstratedin vitroantagonistic activity against different plant pathogenic fungi. Under greenhouse conditions, it was shown that plants infected withRhizoctonia solaniandFusarium verticillioidesand treated withB. subtilis21 produced augment in the number of leaves per plant and an increment in the length of healthy leaves in comparison with untreated plants. In addition,B. subtilis21 showed activity against pathogenic bacteria. Secreted proteins byB. subtilis21 were studied, detecting the presence of proteases and bacteriocin-like inhibitor substances that could be implicated in its antagonistic activity. Chitinases and zwittermicin production could not be detected. Then,B. subtilis21 could potentially be used to control phyt...

Biofilms have been of considerable interest in the context of food hygiene. Of special significance is the ability of microorganisms to attach and grow on food and food-contact surfaces under favourable conditions. Biofilm formation is a dynamic process and different mechanisms are involved in their attachment and growth. Extracellular polymeric substances play an important role in the attachment and colonization of microorganisms to food-contact surfaces. Various techniques have been adopted for the proper study and understanding of biofilm attachment and control. If the microorganisms from food-contact surfaces are not completely removed, they may lead to biofilm formation and also increase the biotransfer potential. Therefore, various preventive and control strategies like hygienic plant lay-out and design of equipment, choice of materials, correct use and selection of detergents and disinfectants coupled with physical methods can be suitably applied for controlling biofilm formation on food-contact surfaces. In addition, bacteriocins and enzymes are gaining importance and have an unique potential in the food industry for the effective biocontrol and removal of biofilms. These newer biocontrol strategies are considered important for the maintenance of biofilm-free systems, for quality and safety of foods.

We have shown previously that in the presence of inducer Bacillus cereus 183, significant increases in bacteriocin production and bactericidal activity of B. thuringiensis occur when the latter is cultivated at pH 7.2, 28°C, and 180 rpm. Here we show that this activity can be further improved when B. thuringiensis is induced with B. cereus 183 and then cultivated with modification of pH, temperature, and agitation. Five native strains of B. thuringiensis, LBIT 269, LBIT 287, LBIT 404, LBIT 420, and LBIT 524 which synthesize, respectively, morricin 269, kurstacin 287, kenyacin 404, entomocin 420, and tolworthcin 524, were cultivated in four different fermentation media. Of these, fermentation in tryptic soy broth (TSB) yielded the highest level of bacteriocin activity (~100–133 FU). Bacteria grown in TSB were induced with B. cereus 183 and cultivated at different pH (6.0, 7.2, 8.0), temperature (26, 28, 30°C), and agitation (150, 180, 210 rpm). Full factorial design was performed and results were analyzed with analysis of variance (ANOVA) and Tukey multiple comparison tests at significant level of α ≤ 0.05 to study the influence of the three variables on bacterial growth and bacteriocin production. Our data show that the highest bacteriocin activity was found with LBIT 269 and LBIT 404 with an increase of ~95–100% compared with induced B. thuringiensis strains cultivated under fixed conditions (pH 7.2, 28°C, 180 rpm), for which the data were set at 0%. The optimal conditions for morricin 269 and kenyacin 404 production were, respectively, pH 8, 30°C, 210 rpm and pH 7.2, 26°C, 210 rpm.

Fermenticin HV6b is a class IIa antimicrobial peptide produced byLactobacillus fermentumHV6b MTCC 10770 isolated from human vaginal ecosystem. It shows growth inhibition of a wide range of opportunistic pathogens of humans, for example,Bacteroides,Gardnerella vaginalis,Mobiluncus,Staphylococci,andStreptococci,associated with bacterial vaginosis in humans. It does possess an impressive sperm immobilization and spermicidal activity tested against human sperms which makes it an attractive proposition for formulating antibacterial vaginosis and contraceptive products. Apart from this,in vitrostudies conducted against four different tissue models have indicated its potential to be used as a component of anticancerous drug therapy as it is reported to induce apoptosis in cancerous cells. This information could be integrated in future studies focusing onin vivoassessment of anticancerous activity of lactic acid bacterial toxins or bacteriocins.

Five psychrotrophic strains of lactic acid bacteria (Lactobacillus casei, Lact. plantarum and Pediococcus spp.) were isolated from 22 samples of commercial salads. These strains were shown to inhibit Aeromonas hydrophila, Listeria monocytogenes, Salmonella typhimurium and Staphylococcus aureus on MRS agar, in salads and in juice prepared from vegetable salads. Lactobacillus casei IMPCLC34 was most effective in reducing total mesophilic bacteria and the coliform group; Aer. hydrophila, Salm. typhimurium and Staph. aureus disappeared after 6 d of storage, while the counts for L. monocytogenes remained constant. The potential application of antimicrobial-producing lactic acid bacteria as biopreservatives of ready-to-use vegetables is suggested.

Approximately 1000 lactobacillus strains were isolated and screened for the production of antimicrobial activity, using a target panel of spoilage organisms and pathogens. Only eight positive strains were found; two of these were studied in more detail. Lactobacillus salivarius M7 produces the new broad spectrum bacteriocin salivaricin B which inhibits the growth of Listeria monocytogenes, Bacillus cereus, Brochothrix thermosphacta, Enterococcus faecalis and many lactobacilli. A new atypical bacteriocin produced by Lact. acidophilus M46, acidocin B, combines the inhibition of Clostridium sporogenes with a very narrow activity spectrum within the genus Lactobacillus and was selected for further characterization. Acidocin B is sensitive to trypsin, heat-stable (80 degrees C for 20 min) and can be extracted from the culture supernatant fluid with butanol. Native acidocin B occurs as a large molecular weight complex (100 kDa), while with SDS-PAGE the partly purified activity migrates as...

M. ZAMFIR, R. CALLEWAERT, P.C. CORNEA, L. SAVU, I. VATAFU and L. DE VUYST.1999.Lactobacillus acidophilus IBB 801 produces a small bacteriocin, designated acidophilin 801, with an estimated molecular mass of less than 6·5 kDa. It displays a narrow inhibitory spectrum (only related lactobacilli but including the Gram-negative pathogenic bacteria Escherichia coli Row and Salmonella panama 1467) with a bactericidal activity. The antimicrobial activity of cell-free culture supernatant fluid was insensitive to catalase but sensitive to proteolytic enzymes such as trypsin, proteinase K and pronase, heat-stable (30 min at 121 °C), and maintained in a wide pH range. The proteinaceous compound was isolated from cell-free culture supernatant fluid and purified. Crude bacteriocin was isolated as a floating pellicle after ammonium sulphate precipitation (40% saturation) and partially purified by extraction/precipitation with chloroform/methanol (2/1, v/v). Further purification to homogeneity was performed by reversed phase Fast Performance Liquid Chromatography. The amino acid composition was determined. Amino acid sequencing revealed that the N-terminal end was blocked.

Enterococci are gram-positive bacteria and fit within the general definition of lactic acid bacteria. Modern classification techniques resulted in the transfer of some members of the genus Streptococcus, notably some of the Lancefield’s group D streptococci, to the new genus Enterococcus. Enterococci can be used as indicators of faecal contamination. They have been implicated in outbreaks of foodborne illness, and they have been ascribed a beneficial or detrimental role in foods. In processed meats, enterococci may survive heat processing and cause spoilage, though in certain cheeses the growth of enterococci contributes to ripening and development of product flavour. Some enterococci of food origin produce bacteriocins that exert anti-Listeria activity. Enterococci are used as probiotics to improve the microbial balance of the intestine, or as a treatment for gastroenteritis in humans and animals. On the other hand, enterococci have become recognised as serious nosocomial pathogens causing bacteraemia, endocarditis, urinary tract and other infections. This is in part explained by the resistance of some of these bacteria to most antibiotics that are currently in use. Resistance is acquired by gene transfer systems, such as conjugative or nonconjugative plasmids or transposons. Virulence of enterococci is not well understood but adhesins, haemolysin, hyaluronidase, aggregation substance and gelatinase are putative virulence factors. It appears that foods could be a source of vancomycin-resistant enterococci. This review addresses the issue of the health risk of foods containing enterococci.

Probiotics including Lactobacillus, Bifidobacteria, Enterococcus, Streptococcus, Bacillus, Lactococcus and some yeasts are usually present in the human gastrointestinal tract, fermented milk and other dairy products, human faeces and breast milk. Source of strains and the ability to reach the target site in the gastrointestinal tract are important. Strains must be able to survive on different physiochemical, enzymatic and microbial stresses throughout the gastrointestinal transit before they can exhibit beneficial effects on the host's health. They can improve the intestinal microbial equilibrium and stimulate butyrate production, which promotes the growth of epithelial cells that lead to an increase in the thickness of the cecal and colonic mucosa for a better absorption of nutrients. They also produce a variety of metabolic end products with antagonistic properties against pathogens. These products include bactericidal proteins and antibiotic-like metabolites termed bacteriocins (such as nisin). The inhibitory spectra of several bacteriocins include food spoilage microorganisms and/or food-borne pathogens. Bacteriocins are considered to be safe natural preservatives or bio-preservatives because it is assumed that they can be degraded by the proteases present in the gastrointestinal tract. The joint working group of Food Aid Organization/World Health Organization made guidelines that can be used for assessing probiotics in food. The minimum requirements include assessment of strain identity and safety, and studies of health benefits in the target host. The continuous search for novel probiotics of importance in medical, industrial and agricultural environments is ongoing around the word. This review summarizes the current state of evaluation criteria for determining the candidacy of novel probiotics.

Biofilms have been of considerable interest in the context of food hygiene. Of special significance is the ability of microorganisms to attach and grow on food and food-contact surfaces under favourable conditions. Biofilm formation is a dynamic process and different mechanisms are involved in their attachment and growth. Extracellular polymeric substances play an important role in the attachment and colonization of microorganisms to food-contact surfaces. Various techniques have been adopted for the proper study and understanding of biofilm attachment and control. If the microorganisms from food-contact surfaces are not completely removed, they may lead to biofilm formation and also increase the biotransfer potential. Therefore, various preventive and control strategies like hygienic plant lay-out and design of equipment, choice of materials, correct use and selection of detergents and disinfectants coupled with physical methods can be suitably applied for controlling biofilm formation on food-contact surfaces. In addition, bacteriocins and enzymes are gaining importance and have an unique potential in the food industry for the effective biocontrol and removal of biofilms. These newer biocontrol strategies are considered important for the maintenance of biofilm-free systems, for quality and safety of foods.