Graciela Vignolo

A review of bacteriocinogenic lactic acid bacteria used as bioprotective cultures in fresh meat produced in Argentina

Several lactic acid bacteria (LAB) associated with meat products are important natural bacteriocin producers. Bacteriocins are proteinaceous antagonistic substances that are important in the control of spoilage and pathogenic microorganisms. The use of LAB as bioprotective cultures to extend the shelf life of fresh meat can improve microbial stability and safety in commercial meat preservation. Lactobacillus curvatus CRL705 used as a protective culture in fresh beef is effective in inhibiting Listeria innocua and Brochothrix thermosphacta as well as the indigenous contaminant LAB, retaining its inhibitory effect at low temperatures and having a negligible effect on meat pH. In addition to the hurdle represented by low temperature and vacuum-packaging, the use of live cells of Lb. curvatus CRL705 seems more feasible from an economic point of view - and without legal restrictions - compared to the addition of purified bacteriocins. A description of meat-borne bacteriocins and their application in meat to extend shelf life is discussed.

Biotechnology of lactic acid bacteria: novel applications.

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Combined Effect of Bacteriocins on the Survival of Various Listeria Species in Broth and Meat System

The antilisterial efficiency of three bacteriocins from lactic acid bacteria, lactocin 705 (produced by L. casei CRL705, 17000 AU/ml), enterocin CRL35 (produced by E. faecium CRL35, 17000 AU/ml), and nisin (2000 IU/ml), was tested in broth, individually and in combination against Listeria monocytogenes and Listeria innocua. Both Listeria species showed an initial decrease in viable counts followed by the regrowth of the survivors after 1 h in the presence of each bacteriocin. A greater antilisterial effect was observed when the bacteriocins were combined in pairs, maximal inhibition being reached when nisin was involved. When a mix of the three bacteriocins was used, no survivors were observed after 24 h of incubation. Similar results were obtained when the bacteriocin combinations were tested in a meat system, indicating that the use of more than one LAB bacteriocin in combination may be effective in preventing the spontaneous emergence of a bacteriocin-resistant Listeria population.

Hydrolysis of muscle myofibrillar proteins by Lactobacillus curvatus and Lactobacillus sake.

Proteolytic enzyme activities of whole cells and cell free extracts (CFE) of Lactobacillus curvatus CECT 904 and Lactobacillus sake CECT 4808 were characterised using synthetic chromogenic compounds and myofibrillar proteins as substrates. The hydrolytic action was monitored by SDS-PAGE and reverse phase-HPLC analyses. The CFE of L. sake partially contributed, together with muscle enzymes, to the initial hydrolysis of myofibrillar proteins. Whole-cells of both L. curvatus and L. sake generated peptides considered important for cured-meat taste. The peptide mapping, resulting from the action on the substrates assayed, revealed a profile of extra and intracellular enzymes. Both strains expressed strong amino acid metabolism.

Control of Listeria monocytogenes in ground beef by ‘Lactocin 705’, a bacteriocin produced by Lactobacillus casei CRL 705

The effectiveness of Lactobacillus casei CRL 705 as well as that of Lactocin 705, the associated bacteriocin produced, in reducing population levels and growth of Listeria monocytogenes in sterile and non-sterile ground beef was studied. Predetermined numbers of L. monocytogenes and concentrations of Lactocin 705 were added to the meat slurries. Listeria in the bacteriocin-treated and control samples during incubation at 20 degrees C were enumerated as CFU on Bacto blood agar base. Results indicated that reduction in numbers of Listeria was largest with high levels of Lactocin 705 and few initial cell numbers of the pathogen present in the meat slurry. However, when the producer strain was added to the slurry, no significant inhibition was detected. Furthermore, inhibition by listeria was shown to be greater when meat slurries were heat-treated.

Role of lactic acid bacteria during meat conditioning and fermentation: Peptides generated as sensorial and hygienic biomarkers

The microbial ecology of meat fermentation is a complex process in which lactic acid bacteria (LAB) and coagulase-negative cocci play a major role. The present work reviews the most significant developments in which LAB are the main characters acting both as starter cultures improving the sensorial quality and as biopreservative agents. New findings about the identification of low molecular weight peptides arisen from protein hydrolysis in dry fermented sausages and their relation with flavor is presented. Also, a brief description of a proteomic approach is detailed in order to exemplify its application as a tool in the search for improved LAB strains that will contribute to food quality and safety. Finally, the most important features of bacteriocinogenic LAB and its bacteriocins in bioprotection of meat and meat products are analyzed.

Modeling lactic acid bacteria growth in vacuum-packaged cooked meat emulsions stored at three temperatures

Abstract The effect of three storage temperatures on the growth of lactic acid bacteria (LAB) in cooked meat emulsions packaged in low oxygen permeability film was investigated. Bacterial counts at 0°C, 8°C and 15°C were fitted to the Gompertz equation and the maximum specific growth rate ( μ ) was obtained as derived parameter, this value being maximal at 15°C (1.16 days −1 ). Arrhenius and root square models were used to describe the effect of different storage temperatures on maximum specific growth rate. The models were statistically validated and the mean square error (MSE), coefficient of determination ( R 2 ), bias factor and accuracy factor were used to evaluate and compare the performance of predictive models. The effect of temperature was better interpreted by root square model than by Arrhenius type model, showing the least deviations from the observed value, which would lead to “fail-safe” prediction of shelf-life. Since the final number of LAB did not show significant changes with storage temperature after 25 days, a temperature abuse during storage will not result in an increase of spoilage by LAB.

Inhibition of Listeria innocua and Brochothrix thermosphacta in vacuum‐packaged meat by addition of bacteriocinogenic Lactobacillus curvatus CRL705 and its bacteriocins

Aims:  To evaluate the inhibition effectiveness of Lactobacillus curvatus CRL705 used as a bioprotective culture and of its bacteriocins, lactocin 705 and lactocin AL705, against Listeria innocua, Brochothrix thermosphacta and indigenous lactic acid bacteria (LAB) in vacuum‐packaged meat stored at 2°C.

Tyramine degradation and tyramine/histamine production by lactic acid bacteria and Kocuria strains

Of 53 strains of lactic acid bacteria and Kocuria, screened for production or degradation of biogenic amines, 29 Kocuria varians and four strains of Enterococcusfaecalisproduced tyramine and, at lower concentrations, histamine. In contrast, Lactobacillus strains that did not possess amino acid decarboxylase activity degraded tyramine. The greatest tyramine oxidase activity was present in the strains L. casei CRL705 (98% degradation) and CRL678 (93%) as well as in L. plantarum CRL681 (69%) and CRL682 (60%).

Mode of action of lactocin 705, a two-component bacteriocin from Lactobacillus casei CRL705.

Lactocin 705 is a bacteriocin whose activity depends on the complementary action of two peptides (705alpha and 705beta) of 33-amino-acid residues each and is produced by Lactobacillus casei CRL705. Biologically active, synthetic lactocin 705 was used to study the mode of action on sensitive cells of Lactobacillus plantarum CRL691. The addition of 90 nmol l(-1) of lactocin 705 to cells of L. plantarum dissipated both, the membrane potential (DeltaPsi) and the pH gradient (DeltapH). Energized membrane, obtained after the addition of glucose, were more susceptible to lactocin 705 action leading to the immediate release of intracellular K(+) and inorganic phosphate. When the role of various ions on sensitive cells were analyzed, only Ca(2+) ion exhibited a protective effect against lactocin 705. These data suggest that the presence of a proton motive force (PMF) promotes the interaction of the bacteriocin with the cytoplasmic membrane of energized cells, leading to pore formation which allows for the efflux of ions, thereby ensuring efficient killing of target bacteria.