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Exocellular polysaccharides produced by lactic acid bacteria.

The production of homopolysaccharides (dextrans, mutans) and heteropolysaccharides by lactic acid bacteria, their chemical composition, their structure and their synthesis are outlined. Mutans streptococci, which include Streptococcus mutans and S. sobrinus produce soluble and insoluble alpha-glucans. The latter may contain as much as 90% alpha-1-3 linkages and possess a marked ability to promote adherence to the smooth tooth surface causing dental plaque. Dextrans produced by Leuconostoc mesenteroides are high molecular weight alpha-glucans having 1-6, 1-4 and 1-3 linkages, varying from slightly to highly branched; 1-6 linkages are predominant. Emphasis is put on exopolysaccharide producing thermophilic and mesophilic lactic acid bacteria, which are important in the dairy industry. The produced polymers play a key role in the rheological behaviour and the texture of fermented milks. One of the main problems in this field is the transitory nature of the thickening trait. This instability is not yet completely understood. Controversial results exist on the sugar composition of the slime produced, but galactose and glucose have always been identified with galactose predominating in most cases.

Dextrans↗

Lactic acid oligomer microspheres containing hydrophilic drugs.

A new method was developed for preparation of biodegradable lactic acid oligomer microspheres containing hydrophilic drugs. The microspheres were obtained by removal of solvent from an O/O (oil-in-oil) emulsion through evaporation. The solvent used for the dispersed phase solution was an acetonitrile: water mixture, while the continuous phase medium was cottonseed oil. Doxorubicin hydrochloride (ADR) and insulin were successfully entrapped in the microspheres with high trapping efficiencies of 80 to 90%, and their release profiles were not accompanied with the significant burst effect. The release rate of the drugs from the microspheres was greatly affected by the initial loading of the drugs and the molecular weight of the lactic acid oligomer.

Aclarubicin↗

Broad and complex antifungal activity among environmental isolates of lactic acid bacteria.

More than 1200 isolates of lactic acid bacteria isolated from different environments were screened for antifungal activity in a dual-culture agar plate assay. Approximately 10% of the isolates showed inhibitory activity and 4% showed strong activity against the indicator mould Aspergillus fumigatus. The antifungal spectra for 37 isolates with strong activity and five isolates with low or no activity were determined. Several of the strains showed strong inhibitory activity against the moulds A. fumigatus, Aspergillus nidulans, Penicillium commune and Fusarium sporotrichioides, and also against the yeast Rhodotorula mucilaginosa. Penicillium roqueforti and the yeasts Pichia anomala and Kluyveromyces marxianus were not inhibited. Several isolates showed reduced antifungal activity after storage and handling. The majority of the fungal inhibitory isolates were identified by 16S rDNA sequencing as Lactobacillus coryniformis. Lactobacillus plantarum and Pediococcus pentosaceus were also frequently identified among the active isolates. The degree of fungal inhibition was not only related to production of lactic or acetic acid. In addition, antifungal cyclic dipeptides were identified after HPLC separation and several other active fractions were found suggesting a highly complex nature of the antifungal activity.

Animals↗

Growth and lactic acid production by vaginal Lactobacillus acidophilus CRL 1259, and inhibition of uropathogenic Escherichia coli.

Lactic acid-producing lactobacilli were selected from 134 human vaginal isolates by testing their capability to inhibit the growth of different pathogenic micro-organisms. Lactobacillus acidophilus CRL 1259 (from the CERELA Culture Collection) was selected to study the effects of temperature, pH and culture medium on growth and lactic acid production. Growth parameters were estimated by using the model of Gompertz. Kinetics of inhibition of uropathogenic Escherichia coli were evaluated in mixed cultures of the pathogen and L. acidophilus. Optimal conditions for growth and lactic acid production by L. acidophilus were pH 6.5 or 8.0 and 37 degrees C. Under these conditions, growth was higher in LAPTg (yeast extract/peptone/tryptone/Tween 80/glucose) broth than in MRS (De Man-Rogosa-Sharpe) broth. However, lactic acid production was more efficient in MRS broth. Under optimal conditions for lactic acid production, L. acidophilus inhibited the growth of E. coli. These results suggest that inclusion of L. acidophilus CRL 1259 in probiotic products for vaginal application would be beneficial.

Adult↗

Effect of systemic pH on pHi and lactic acid generation in exhaustive forearm exercise.

To investigate whether changes in systemic pH affect intracellular pH (pHi), energy-rich phosphates, and lactic acid generation in muscle, eight normal volunteers performed exhaustive forearm exercise with arterial blood flow occluded for 2 min on three occasions. Subjects ingested 4 mmol/kg NH4Cl (acidosis; A) or NaHCO3 (alkalosis; B) or nothing (control; C) 3 h before the exercise. Muscle pHi and phosphocreatine (PCr) content were measured with 31P-nuclear magnetic resonance (31P-NMR) spectroscopy during exercise and recovery. Lactate output during 0.5-7 min of recovery was calculated as deep venous-arterial concentration differences times forearm blood flow. Before exercise, blood pH and bicarbonate were lower in acidosis (7.303 +/- 0.009, 18.6 +/- 0.5 meq/l) than alkalosis (7.457 +/- 0.010, 32.2 +/- 0.7 meq/l) and intermediate in control (7.389 +/- 0.007, 25.3 +/- 0.6 meq/l). Lactic acid output during recovery was less with A (245 +/- 39 mumol/100 ml) than B (340 +/- 55 mumol/100 ml) (P less than 0.05) and intermediate in C (293 +/- 31 mumol/100 ml). PCr utilization and resynthesis were not affected by extracellular pH changes. pHi did not differ before exercise (A, 7.04 +/- 0.01; B, 7.09 +/- 0.01; C, 7.06 +/- 0.01) or at its end (A, 6.28 +/- 0.07; B, 6.28 +/- 0.11; C, 6.31 +/- 0.09). Hence systemic acidosis inhibited and alkalosis stimulated lactic acid output. These findings suggest that systemic pH regulates cellular acid production, protecting muscle pH, at the expense of energy availability.

Adult↗

[Comparison of methods of storing lactic acid bacteria].

The study was undertaken to elucidate how different methods of storage (immersing in mineral oil, lyophilization, and subculturing) of lactic acid bacteria belonging to the genera Lactobacillus and Lactococcus affect their viability, antibiotic activity, and ability to accumulate organic acids. Storage of the lactic acid bacterium L. lactis subsp. lactis by immersion in mineral oil proved to be ineffective. Lyophilization allowed the survival of a sufficiently large number of cells, although their antibiotic activity somewhat decreased. The resuscitation of lyophilized bacteria by subculturing them in rich nutrient media, such as skim milk, led to the restoration of their physiological activity, including the effective antimicrobial spectrum.

Culture Media↗

Dissociation of gas-phase dimeric complexes of lactic acid and transition-metal ions formed under electrospray ionization conditions; the role of reduction of the metal ion.

Dimeric complex ions of the type [M(A-H)A]+, where M=metal ion (Co, Ni, Cu, and Zn) and A=ligand (lactic acid, methyl lactate or ethyl lactate), were generated in the gas phase under electrospray ionization conditions. The collision-induced dissociation spectra of [M(A-H)A]+ ions were recorded to study the behaviour of ligand and metal ions in decomposition of these dimeric complex ions. Based on the fragmentation pathways observed for complex ions of lactic acid, it is found that both the carboxylic and hydroxyl groups of lactic acid are involved in the complex formation following displacement of a proton by the metal ion. The dimeric complex ions of Co, Ni, and Zn dissociated to yield similar types of ions, whereas that of Cu behaved differently. The dissociations of Co-, Ni-, and Zn-bound dimeric complexes involved losses of neutral molecules while keeping the oxidation state of the metal ion unchanged. However, elimination of radicals is found in the dissociation of dimeric complex ions of Cu, and the oxidation state of copper is reduced from Cu(II) to Cu(I) in the resulting fragment ions. The deprotonated ligand is involved in the fragmentation pathway of Cu complexes, whereas it is intact in other complexes. The oxidation state of the metal ion, nature of the ligand, and site of attachment to the metal ion are found to control the dissociation of these dimeric complex ions.

Dimerization↗

Nicotinamide adenine dinucleotide-dependent and nicotinamide adenine dinucleotide-independent lactate dehydrogenases in homofermentative and heterofermentative lactic acid bacteria.

Three homofermentative (Lactobacillus plantarum B38, L. plantarum B33, Pediococcus pentosaceus B30) and three heterofermentative (Leuconostoc mesenteroides 39, L. oenos B70, Lactobacillus brevis) lactic acid bacteria were examined for the presence or absence of nicotinamide adenine dinucleotide (NAD)-dependent and NAD-independent d- and l-lactate dehydrogenases. Two of the six strains investigated, P. pentosaceus and L. oenos, did not exhibit an NAD-independent enzyme activity capable of reducing dichlorophenol indophenol. The pH optima of the lactic dehydrogenases were determined. The NAD-dependent enzymes from homofermentative strains exhibited optima at pH 7.8 to 8.8, whereas values from 9.0 to 10.0 were noted for these enzymes from heterofermentative organisms. The optima for the NAD-independent enzymes were between 5.8 and 6.6. The apparent Michaelis-Menten constants determined for both NAD and the substrates demonstrated the existence of a greater affinity for d- than l-lactic acid. A comparison of the specific NAD-dependent and NAD-independent lactate dehydrogenase activities revealed a direct correlation of the d/l ratios of these activities with the type of lactic acid produced during the growth of the organism.

Ammonium Sulfate↗

Thermogelling aqueous solutions of alternating multiblock copolymers of poly(L-lactic acid) and poly(ethylene glycol).

We are reporting alternating multiblock copolymers of poly(L-lactic acid)/poly(ethylene glycol) aqueous solution (> 15 wt %) undergoing sol-gel-sol transition as the temperature increases from 20 to 60 degrees C. Micelles of the multiblock copolymers (in water) are about 20 nm in radius at low temperature. They are aggregated to a larger size as the temperature increases, which should play a critical role in the sol-to-gel transition. The transition temperature and gel window were affected by the molecular weight and composition of the multiblock copolymer. In particular, the aqueous solution of an alternating multiblock copolymer (Mn approximately 6700 daltons) prepared from poly(ethylene glycol) (Mn approximately 600 daltons) and poly(L-lactic acid) (Mn approximately 1300 daltons) showed a maximum modulus at body temperature (37 degrees C). The in situ gel forming ability of the polymer aqueous solution in vivo as well as in vitro indicates that it can be a promising injectable biomaterial.

Animals↗

The value of peritoneal fluid hypoxanthine besides lactic acid in detection of the vascular compromise of intestine.

Hypoxanthine has been shown to be a sensitive and more specific measure of hypoxia than lactate in several body fluids other than peritoneal fluid. An experimental study was conducted to determine the value of peritoneal fluid hypoxanthine besides lactic acid levels in detecting vascular compromise of the intestine. Thirty male guinea pigs were divided in 3 groups each containing 10 animals. Groups comprised of sham operation control, simple intestinal obstruction and intestinal obstruction with vascular compromise. Lactic acid and hypoxanthine levels were determined in peritoneal lavage effluents. Peritoneal fluid lactic acid levels increased after 4 hours of intestinal obstruction with vascular compromise (p < 0.01) but not in simple intestinal obstruction and sham operation control groups. No detectable amount of hypoxanthine could be found in any of the fluid samples. It is concluded that unlike in other body fluids hypoxanthine is not present in peritoneal fluid and does not become evident after vascular compromise of the intestine.

Animals↗

Inhibition, survival and growth of Listeria monocytogenes on poultry as influenced by buffered lactic acid treatment and modified atmosphere packaging.

The effect of the treatment with various concentrations (2%, 5% and 10% w/v) of lactic acid/sodium lactate buffer (pH 3.0), modified atmosphere (MAP) packaging (90% CO2 and 10% O2) and 10% (w/v) lactic acid/sodium lactate buffer (pH 3.0) combined with MAP on Listeria monocytogenes Z7 serotype 1 and on the shelf life of chicken legs stored at 6 degrees C was investigated. The initial contamination level of L. monocytogenes on the chicken legs surface was 8.3 x 10(2) cfu/cm2 of skin. After 2 days of storage at 6 degrees C the number of L. monocytogenes on legs treated with 2%, 5%, 10% lactic acid/sodium lactate buffer (pH 3.0) and 10% lactic acid/sodium lactate buffer (pH 3.0) combined with MAP was significantly lower than the initial number of L. monocytogenes. Later, growth of L. monocytogenes was observed. After 13 days of storage at 6 degrees C the number of L. monocytogenes on legs treated with 10% lactic acid/sodium lactate buffer (pH 3.0) combined with MAP was still similar to the initial number. Legs treated with 2%, 5%, 10% lactic acid/sodium lactate buffer (pH 3.0), MAP and 10% lactic acid/sodium lactate buffer (pH 3.0) combined with MAP, have a shelf life at 6 degrees C of respectively 8, 9, 10, 13 and 17 days. This means a prolongation of 2, 3, 4, 7 and 11 days, respectively for storage at 6 degrees C. The antimicrobial effect of lactic acid buffer systems (pH 3.0) increased with increasing concentrations of lactic acid in the buffered system. The best results were obtained by the combined use of 10% acid/sodium lactate buffer (pH 3.0) and MAP.

Analysis of Variance↗

Influence of immobilization parameters on growth and lactic acid production by Streptococcus thermophilus and Lactobacillus bulgaricus co-immobilized in calcium alginate gel beads.

Streptococcus thermophilus and Lactobacillus bulgaricus were co-immobilized in different systems with varying calcium (0.1-1.5 M) and alginate (1-2%, w/v) concentrations. Highest lactic acid production was 35 g l(-1) when both bacteria were in high viscosity beads (1%, w/v alginate) hardened in 0.1 M CaCl2 . The gel bead composition affected size and distribution of entrapped lactic acid bacteria.

Alginates↗

Modelling the growth limits (growth/no growth interface) of Escherichia coli as a function of temperature, pH, lactic acid concentration, and water activity.

The form of a previously developed Bĕlehrádek type of growth rate model was used to develop a probability model for defining the growth/no growth interface as a function of temperature (10 to 37 degrees C), pH (pH 2.8 to 6.9), lactic acid concentration (0 to 500 mM), and water activity (0.955 to 0.999; NaCl was used as the humectant). Escherichia coli was unable to grow in broth in which the undissociated lactic acid concentration exceeded 11 mM or, with two exceptions, at a pH of 3.9 or less with no lactic acid present. Under experimental conditions at which the pH and the undissociated acid concentrations were the major growth-limiting factors, the growth/no growth interface was essentially independent of temperature at temperatures ranging from 15 to 37 degrees C. The interface between conditions that allowed growth and conditions at which growth did not occur was abrupt. The inhibitory effect of combinations of water activity and pH varied with temperature. Predictions of the model for the growth/no growth interface were consistent with 95% of the experimental data set.

Escherichia coli↗

Cholesterol assimilation by lactic acid bacteria and bifidobacteria isolated from the human gut.

The objective of this study was to evaluate the effect of human gut-derived lactic acid bacteria and bifidobacteria on cholesterol levels in vitro. Continuous cultures inoculated with fecal material from healthy human volunteers with media supplemented with cholesterol and bile acids were used to enrich for potential cholesterol assimilators among the indigenous bacterial populations. Seven potential probiotics were found: Lactobacillus fermentum strains F53 and KC5b, Bifidobacterium infantis ATCC 15697, Streptococcus bovis ATCC 43143, Enterococcus durans DSM 20633, Enterococcus gallinarum, and Enterococcus faecalis. A comparative evaluation regarding the in vitro cholesterol reduction abilities of these strains along with commercial probiotics was undertaken. The degree of acid and bile tolerance of strains was also evaluated. The human isolate L. fermentum KC5b was able to maintain viability for 2 h at pH 2 and to grow in a medium with 4,000 mg of bile acids per liter. This strain was also able to remove a maximum of 14.8 mg of cholesterol per g (dry weight) of cells from the culture medium and therefore was regarded as a candidate probiotic.

Bifidobacterium↗

Biofilm formation and contamination of cheese by nonstarter lactic acid bacteria in the dairy environment.

Defects in cheese, such as undesirable flavors, gas formation, or white surface haze from calcium lactate crystals, can result from growth of nonstarter lactic acid bacteria (NSLAB). The potential for biofilm formation by NSLAB during cheese manufacturing, the effect of cleaning and sanitizing on the biofilm, and bacterial growth and formation of defects during ripening of the contaminated cheese were studied. Stirred-curd Cheddar cheese was made in the presence of stainless steel chips containing biofilms of either of two strains of erythromycin-resistant NSLAB (Lactobacillus curvatus strain JBL2126 or Lactobacillus fermentum strain AWL4001). During ripening, the cheese was assayed for total lactic acid bacteria, numbers of NSLAB, and percentage of lactic acid isomers. Biofilms of L. curvatus formed during cheese making survived the cleaning process and persisted in a subsequent batch of cheese. The starter culture also survived the cleaning process. Additionally, L. curvatus biofilms present in the vat dislodged, grew to high numbers, and caused a calcium lactate white haze defect in cheese during ripening. On the other hand, biofilms of L. fermentum sloughed off during cheese making but could not compete with other NSLAB present in cheese during ripening. Pulsed-field gel electrophoresis results verified the presence of the two biofilm strains during cheese making and in the ripening cheese. Probable contamination sites in the plant for other NSLAB isolated in the cheese were identified, thus supporting the hypothesis that resident NSLAB biofilms are a viable source of contamination in the dairy environment.

Biofilms↗

Isolation of lactic acid bacteria from Malaysian foods and assessment of the isolates for industrial potential.

Two traditional fermented food 'tapai' (fermented tapioca) and 'tempoyak' (fermented durian flesh), chilli puree and fresh goat's milk were used as sources for the isolation of lactic acid bacteria (LAB). A total of 126 isolates were obtained and by sequential screening for catalase activity and Gram-staining, 55 were determined to be LAB out of which 16 were established to be homofermentative by the gel plug test. Seven isolates were identified by use of the API 50CHL kit and two lactobacilli strains and one lactococci strain were selected to study their growth and lactic acid production profiles in a time course experiment. The lactobacilli strains, both isolated from 'tapai', produced higher amounts of cells and lactic acid from glucose as compared to the lactococci strain isolated from fresh goat's milk.

Animals↗

Use of virginiamycin to control the growth of lactic acid bacteria during alcohol fermentation.

The antibiotic virginiamycin was investigated for its effects on growth and lactic acid production by seven strains of lactobacilli during the alcoholic fermentation of wheat mash by yeast. The lowest concentration of virginiamycin tested (0.5 mg Lactrol kg-1 mash), was effective against most of the lactic acid bacteria under study, but Lactobacillus plantarum was not significantly inhibited at this concentration. The use of virginiamycin prevented or reduced potential yield losses of up to 11% of the produced ethanol due to the growth and metabolism of lactobacilli. However, when the same concentration of virginiamycin was added to mash not inoculated with yeast, Lactobacillus rhamnosus and L. paracasei grew after an extensive lag of 48 h and L. plantarum grew after a similar lag even in the presence of 2 mg virginiamycin kg-1 mash. Results showed a variation in sensitivity to virginiamycin between the different strains tested and also a possible reduction in effectiveness of virginiamycin over prolonged incubation in wheat mash, especially in the absence of yeast.

Anti-Bacterial Agents↗

Use of gas-liquid chromatography to determine the end products of growth of lactic Acid bacteria.

A simple gas-liquid chromatographic procedure for analyzing ethanol, acetic acid, acetoin, and racemic and meso-2,3-butylene glycol in broth media is described. Overnight broth cultures were filtered or centrifuged, and the filtrate or supernatant was treated with formic acid to aid separation of volatile fatty acids. Samples were then directly analyzed by gas-liquid chromatography on a 20% Tween 80-Chromosorb W-AW column and propionic acid as an internal standard. A complete analysis took ca. 8 min. The method can be used to distinguish homofermentative from heterofermentative lactic acid bacteria based on the level of ethanol produced and citrate-utilizing from non-citrate-utilizing lactic acid bacteria based on the levels of acetic acid produced. The method also has potential in distinguishing other bacterial fermentations. Of the 13 species of lactic acid bacteria tested, Streptococcus lactis subsp. diacetylactis was the major producer of 2,3-butylene glycol (total range, 0.3 to 3.5 mM), and, except for strain DRC1, both the racemic and meso isomers were produced in approximately equal amounts.

Journal Article↗