Flow Cytometric Assessment of Viability of Lactic Acid Bacteria

The viability of lactic acid bacteria is crucial for their applications as dairy starters and as probiotics. We investigated the usefulness of flow cytometry (FCM) for viability assessment of lactic acid bacteria. The esterase substrate carboxyfluorescein diacetate (cFDA) and the dye exclusion DNA binding probes propidium iodide (PI) and TOTO-1 were tested for live/dead discrimination using a Lactococcus, a Streptococcus, three Lactobacillus, two Leuconostoc, an Enterococcus, and a Pediococcus species. Plate count experiments were performed to validate the results of the FCM assays. The results showed that cFDA was an accurate stain for live cells; in exponential-phase cultures almost all cells were labeled, while 70°C heat-killed cultures were left unstained. PI did not give clear live/dead discrimination for some of the species. TOTO-1, on the other hand, gave clear discrimination between live and dead cells. The combination of cFDA and TOTO-1 gave the best results. Well-separated subpopulations of live and dead cells could be detected with FCM. Cell sorting of the subpopulations and subsequent plating on agar medium provided direct evidence that cFDA labels the culturable subpopulation and that TOTO-1 labels the nonculturable subpopulation. Applied to cultures exposed to deconjugated bile salts or to acid, cFDA and TOTO-1 proved to be accurate indicators of culturability. Our experiments with lactic acid bacteria demonstrated that the combination of cFDA and TOTO-1 makes an excellent live/dead assay with versatile applications.     

Effect of Compatible Solutes on Survival of Lactic Acid Bacteria Subjected to Drying

Four strains of lactic acid bacteria were investigated to determine if a relationship exists between accumulation of compatible solutes and the ability of cells to survive drying. Betaine was the major solute found in these lactic acid bacteria subjected to salt stress. Survival of cultures subjected to drying was considerably enhanced when this solute was accumulated by cells.     

猪源乳酸菌产乳酸及其抑菌特性研究

研究了5株(L1、12、L3、L5和L7)分离自仔猪肠道的乳酸菌的产乳酸能力及抑菌特性。结果表明：L5菌株产乳酸的速度最快,培养液中乳酸含量最高,L5菌株培养液pH值的下降速度最快,终末pH值最低,而L1菌株产乳酸的速度最慢,培养液乳酸含量最低。5株乳酸菌对大肠杆菌K88、K99、987P、O141和大肠杆菌E1及金黄色葡萄球菌均有不同程度的抑制作用;排除酸的影响后仍有22％～53％抑菌效果;经热处理后保持有92％以上的抑菌效果;蛋白酶处理后保持85％以上的抑菌效果。     

Development of Freeze-Dried Bacteriocin-Containing Preparations from Lactic Acid Bacteria to Inhibit Listeria monocytogenes and Staphylococcus aureus

There has been a recent movement to produce and consume ??minimally processed?? and more ??natural?? foods through the use of fewer chemical preservatives. The shift to more ??natural?? foods has resulted in a great interest in the use of bacteriocins from lactic acid bacteria as natural biopreservatives. The objective of this comparative study was to identify bacteriocins that can be produced in low-cost or no-cost dairy-based media (DBM), concentrated using freeze-drying, and applied to Cheddar cheese samples to concurrently inhibit Listeria monocytogenes and Staphylococcus aureus. Select bacteriocin producers were grown in DBM, their cell-free supernatants (CFS) were frozen, and the frozen CFS samples were freeze-dried to produce bacteriocin-containing powders. Cheddar cheese samples were challenged with L. monocytogenes or Staph. aureus cells. The challenged samples were exposed to buffered solutions of freeze-dried powders containing bacteriocins, incubated at 4?°C for 24?C72?h, and plated onto appropriate selective media. All freeze-dried bacteriocin-containing powders tested were active against L. monocytogenes and Staph. aureus. Our research findings indicated that low-cost or no-cost DBM could successfully be used for production of bacteriocin-containing preparations. In addition, freeze-drying was determined to be a feasible approach to prepare concentrated and stable bacteriocin-containing powders for prospective food applications. The prevention of even a very small percentage of foodborne illnesses via the use of bacteriocins as natural biopreservatives would help reduce the number of foodborne illness-related hospitalizations, deaths, and financial loss due to medical expenses, lost income/productivity, cost of litigation/penalties, and loss of trade.     

Medium for the Selective Enumeration of Lactic Acid Bacteria from Foods

A nitrite actidione polymyxin agar was developed for the enumeration of lactic acid bacteria. It was effective in recovering organisms from pure cultures and from foods.     

Malt Extract as Protective Medium for Lactic Acid Bacteria in Cryopreservation

S ummary : Malt extracts prepared from barley and sorghum malts were used as media for preserving bacteria at -196°. The results are compared with those obtained with other protective substances such as dimethylsulphoxide, glycerol, malic acid, sucrose and sodium glutamate. The malt extract medium is recommended for use in the food industry.     

乳酸菌酸胁迫反应机制研究进展

乳酸菌可发酵糖类产生乳酸,并广泛应用于食品、药物和饲料等工业。由于有机酸的积累,乳酸菌大部分的生长代谢都在低pH的酸性环境中进行,具有酸胁迫反应。pH的自我平衡、ATR反应机制、对大分子的保护和修复作用及细胞膜的变化等是乳酸菌酸胁迫反应的主要机制,其中,pH自我平衡包括F0F1-ATPase质子泵、精氨酸脱氨酶途径(ADI)和谷氨酸脱羧酶途径(GAD)等。由此可见,乳酸菌酸胁迫反应机制涉及到基因和蛋白的表达调控等,是非常复杂的网络调控体系。     

Protective Effect of Adonitol on Lactic Acid Bacteria Subjected to Freeze-Drying

The protective effects of glycerol, adonitol, and four other related polyhydric alcohols on lactic acid bacteria subjected to freeze-drying were examined. The presence of adonitol in the suspending medium markedly protected the viabilities of the 12 stains tested. Dulcitol, mannitol, m-inositol, and sorbitol were found to provide little or no protection.     

Antagonism of Lactic Acid Bacteria against Phytopathogenic Bacteria

A variety of lactic acid bacteria, isolated from plant surfaces and plant-associated products, were found to be antagonistic to test strains of the phytopathogens Xanthomonas campestris, Erwinia carotovora, and Pseudomonas syringae. Effective “in vitro” inhibition was found both on agar plates and in broth cultures. In pot trials, treatment of bean plants with a Lactobacillus plantarum strain before inoculation with P. syringae caused a significant reduction of the disease incidence.     

Antioxidant Properties of Lactic Acid Bacteria

Microbiology - Lactic acid bacteria (LAB) are widely used in fermentation processes for the preparation of various foodstuffs, including dairy, meat and vegetable products. In the course of...     

Effect of Sodium Nitrite and Sodium Chloride on Growth of Lactic Acid Bacteria

The effect of NaNO2 and NaCl on the growth of 24 lactic acid bacteria strains isolated from vacuum-packed cooked ring sausages were examined by analyzing different growth parameters with Bioscreen. NaNO2 had a very limited effect on the growth of lactic acid bacteria at 50 and 100 mg/l but at 400 mg/l a more pronounced inhibitory effect was found. Bacterial growth was enhanced by 1–2% (w/v) of added NaCl, while NaCl concentrations above 3% (w/v) had a clear inhibitory effect. Leuconostoc isolates seemed to be more sensitive to sodium nitrite and sodium chloride than homofermentative lactobacilli strains. Among homofermentative lactobacilli, the strains resembling Lactobacillus curvatus were more sensitive to NaCl than those resembling Lactobacillus sake.     

乳酸菌在基因工程领域的应用

乳酸菌是工业微生物中一类非常重要的细菌,也是众所周知的维持人及动物健康的重要益生菌群.在最近十几年来,乳酸菌的遗传学和分子生物学特性很受关注,通过对它们进行遗传修饰改造取得了重要成果,扩大了它们的应用.     

Studies on Lipolysis of Dairy Lactic Acid Bacteria

Cell-free extracts were prepared by Hughes’ type disintegrator from cells of Lactobacillus casei, L. plantarum, L. helveticus and Streptococcus diacetilactis. Lipolytic activities of these extracts were measured by titration of total acids liberated from tributyrin and butterfal emulsions at given temperature. More acids were liberated from tributyrin than from butterfat, olive oil and composite butter. Lipolytic activities of all extracts for tributyrin were approximately the same at reaction temperature of 30°C, 37°C and 45°C, and were higher at pH 6 to 8. Free fatty acids of C₁₀, C₁₂, C₁₄, C₁₆ and C₁₈ liberated by extracts of L. casei and L. plantarum, were tentatively identified on gas-liquid chromatograms.     

Neuroprotective Effect of Riboflavin Producing Lactic Acid Bacteria in Parkinsonian Models

Neurochemical Research - Oxidative stress and inflammatory processes might contribute to the cascade of events leading Parkinson disease (PD); and vitamins such as riboflavin can exert protection...     

乳酸菌细菌素的研究进展

简要介绍了乳酸菌细菌素的概念和分类,并以nisin为例对其分子结构、理化性质、抑菌范围及机制、遗传控制作了较为详尽的介绍,简要概括了乳酸菌细菌素的应用和今后的研究方向。     

Potentials of Exopolysaccharides from Lactic Acid Bacteria

Recent research in the area of importance of microbes has revealed the immense industrial potential of exopolysaccharides and their derivative oligosaccharides from lactic acid bacteria. However, due to lack of adequate technological knowledge, the exopolysaccharides have remained largely under exploited. In the present review, the enormous potentials of different types of exopolysaccharides from lactic acid bacteria are described. This also summarizes the recent advances in the applications of exopolysaccharides, certain problems associated with their commercial production and the remedies.     

Studies on the Pentose Isomerases of Lactic Acid Bacteria

Production of d-xylose and l-arabinose isomerases by lactic acid bacteria was greatly promoted by the addition of manganese ions in cultural medium. Effective concentration of the ions was 5 × 1O^-3 m. Ferrous ions were also effective for the production of d-xylose isomerase and cobaltous ions were somewhat effective for the production of l-arabinose isomerase. Zinc and cadmium ions inhibited bacterial growth. It was possible to increase the production of isomerase by changing MnSO₄ concentration to 5× 10^-3 m (0.l1 %) in place of 0.001 per cent in the normal medium.

Column chromatographic procedures for the purification of pentose isomerases were carried out. Cation and anion exchange resins were not suitable because of their low exchange capacities and instability of the enzyme at acidic pH range. But the isomerases were successfully purified by DEAE-cellulose column chromatography with high recovery (85~90%). Using a Tris buffer, KCl concentration was increased in gradient. d-Xylose isomerase was eluted at pH 7.0 at 0~0.2 m KCl, and l-arabinose isomerase at pH 8.0 at 0~0.4 m KCl. The purified isomerases, d-xylose isomerase and l-arabinose isomerase, both required manganese ions specifically for their activities.

D-Xylose isomerase and l-arabinose isomerase are different enzymes which can be separated from each other with acetone fractionation at pH 4.8~5.0, heat treatment or chromatography on a colnmn of DEAE-cellulose. In DEAE-cellulose chromatography with a linear gradient elution method, d-xylose isomerase is recovered in the first peak at pH 7.0 (Tris bnffer) with 0~0.2 m KCl, and l-arabinose isomerase is eluted in the second peak at pH 8.0 (Tris buffer) with a larger ionic strength.