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Effects of Lactobacillus casei on Pseudomonas aeruginosa infection in normal and dexamethasone-treated mice.

A single intraperitoneal injection of Lactobacillus casei YIT 0003 into normal or dexamethasone-treated mice led to nonspecific resistance against intraperitoneal challenge with lethal doses of Pseudomonas aeruginosa PAO 3047. The enhanced resistance was retained for 14 days (P less than 0.05) after injection with living L. casei. In contrast, the statistically significant duration of the enhanced resistance in mice treated intraperitoneally with living L. acidophilus YIT 0075 was only 5 days. The in vivo killing activity of peritoneal exudate cells (PECs) against P. aeruginosa 5 and 7 days after intraperitoneal injection of living L. casei was significantly higher than in the case of PECs elicited by L. acidophilus. In the case of intravenous injection of heat-killed L. casei before intraperitoneal challenge with P. aeruginosa, there were no survivors in the late period after administration of L. casei. A high correlation existed between the patterns of in vivo killing of P. aeruginosa by PECs and survival rate of mice injected intravenously with heat-killed L. casei. The reduced in vivo killing activity of PECs from dexamethasone-treated mice against P. aeruginosa infection was also augmented by the intraperitoneal injection of heat-killed L. casei. These results indicate that L. casei possesses a resistance-enhancing capacity against P. aeruginosa infection in vivo. Differences in the duration of the enhanced resistance caused by L. casei and by L. acidophilus may be due to differences in chemical composition and/or physicochemical properties of the cell walls of the two kinds of bacteria.

Animals↗

[Antifungal effect in selected natural compounds and probiotics and their possible use in prophylaxis of vulvovaginitis].

OBJECTIVE: a survey of effective natural compounds and probiotics with antifungal activity and their possible use in vulvovaginitis as a prophylactic or therapeutic drug. DESIGN: review of literature. SETTING: Charles University at Prague, Pharmaceutical Faculty at Hradec Kralove. METHODS: and review of literature and publications from medical data-bases. CONCLUSIONS: certain natural compounds were proved to exert antimycotic effects. The efficiency of garlic extract under in vitro conditions against yeast of C. albicans family becomes manifest within one hour. Essential oils present in the Cayenne pepper (Capsodum frutescens) exert considerable antifungal activity against C. albicans as well. Clove exerts lower antimycotic effects. The effects of Chinese leek are similar to garlic. Propolis also possesses antifungal properties, and sensitivity of the yeast strains decreases in the following order: C. albicans > C. tropicalis > C. crusei > C. guilliermondii. Its efficiency depends on the composition and place of origin. Tea trea oil (Melalauca alternifolia) is inhibiting strains of Candida family (genus) in minimal inhibiting concentration, higher concentrations are necessary for C. crusei and C. norvengensis. In the prophylaxis of vaginal candidoses, oral administration of the strains of Lactobacilus rhamnosus GR-1 and L. fermentum RC-14, B-54 may be recommended. Moreover there are suitable strains of L. brevis CD-2, L. salivarius FV-2, L. gasseri MB 335, L. plantarum, useful in the form of gel capsules (L. plantarum) or vaginal tablets (L. gasseri). The strains of L. acidophilus CRL 1294 and L. salivarius CRL 1328 display the capacity of self aggregation and co-aggregation with Candida. It should be pointed out that the probiotic strain of Lactobacillus (especially L. casei and L. rhamnosus) may, in exceptional cases, cause bacteriemia and sepsis, especially in immunodeficient patients and those who underwent surgery of alimentary tract. No protective effect against yeast infection was detected with vitamin E, any positive effect was not unequivocally confirmed with zinc, but adjusting zinc levels to normal may be recommended.

Antifungal Agents↗

[Protein fingerprinting as a method for strain-specific differentiation of technologically useful Lactobacillus strains from clinical isolates].

Lactobacilli are often used in products of animal origin because of their technological properties and as protective cultures against pathogenic microorganisms, however they are supposed to be related to human infections as well. Therefore 39 strains of species that are in biotechnological usage, including the type strains, and some clinical isolates were investigated. 9 strains of L. acidophilus, 7 of L. gasseri and 23 strains of L. rhamnosus were tested. Biochemical and physiological properties were proved by classical tests. Analysis of the total soluble cytoplasmatic protein patterns was performed with diamine silver staining, a technique not previously applied to lactobacilli. Classical tests were able to confirm all strains as belonging to one of the three species. Analysis of protein patterns allowed to differentiate between strains of the same species. In case of L. acidophilus biotechnologically used strains and clinical isolates were clustered separately. L. rhamnosus formed three clusters. SDS-PAGE of proteins together with diamine silver staining seems to be helpful to detect intra-species differences. It was stated, that technologically used strains of lactobacilli could clearly be differentiated from another and also from clinical isolates.

Animals↗

Antimutagenicity and the influence of physical factors in binding Lactobacillus gasseri and Bifidobacterium longum cells to amino acid pyrolysates.

Antimutagenic and binding properties of 28 strains of Lactobacillus gasseri and 2 strains of Bifidobacterium longum on the mutagenicity of amino acid pyrolysates were investigated in vitro using a streptomycin-dependent (SD510) strain of Salmonella typhimurium TA 98. Four strains of L. acidophilus (SBT0274, SBT1703, SBT10239, and SBT10241) and 1 strain of B. longum (SBT 2928) exhibited the highest percentage of antimutagenicity and binding. These 5 strains were further optimized for other physical factors influencing the mechanism of binding, such as cell and mutagen concentration, pH, and incubation time. In all of the selected strains, 2 mg of cells bound with 88 to 95% of 0.2 mg of 3-amino-1,4 dimethyl-5H-pyrido[4,3-b]indole in 30 min at pH 7.0. Other amino acid pyrolysates, such as 3-amino-1-methyl-5H-pyrido[4,3-b]indole, 2-amino-6-methyldi-pyrido[1,2-a:3',2'-d]imidazole, 2-amino-3-methyl-imidazo[4,5,f]quinoline, and 2-amino-3,4-dimethyl-imidazo[4,5,f]quinoline were also tested for the binding ability of these strains. We observed that the complexity of the mutagens greatly influenced the binding properties. The binding of 3-amino-1,4 dimethyl-5H-pyrido[4,3-b]indole to the purified cell walls was very high compared with that of the crude cell wall, peptidoglycan, or the cell extract. Binding was inhibited when the cell walls were subjected to treatment with metaperiodate or trichloroacetic acid but not when they were subjected to treatment with lysozyme, trypsin, or proteinase K, reflecting the role of the carbohydrate component as a binding site.

Amino Acids↗

Identification of Lactobacillus species using tDNA-PCR.

tDNA intergenic spacer PCR (tDNA-PCR) using consensus primers complementary to the conserved edges of the tRNA genes can amplify the intergenic spacers. Separation of the PCR products with capillary electrophoresis enables discrimination between fragments differing only one basepair in length. This method was applied to a collection of 82 Lactobacillus strains belonging to 37 species in order to evaluate the discriminatory power of this technique within this genus. Twenty-one species could be distinguished to species level on the basis of a unique tDNA fingerprint pattern. The other species grouped by two (e.g. L. fermentum and L. cellobiosus) or three (L. acidophilus, L. gallinarum and L. helveticus). Inclusion of the resulting fingerprints in a numerical database containing fingerprints of numerous other Gram-positive and Gram-negative species makes the identification of unknown strains possible.

Bacterial Typing Techniques↗

Detection of caries-inducing microorganisms in hyposalivated rats without infection of mutans streptococci.

Rampant dental caries was induced in hyposalivated rats fed a high sucrose diet without infection of mutans streptococci, in which increased numbers of lactobacilli and S. aureus were demonstrated in the oral flora. Administration of either penicillin or piperacillin, effective against all isolates of lactobacilli, markedly inhibited the caries induction in these rats, while severe dental caries was induced in hyposalivated rats given vancomycin that is inhibitory against S. aureus. These results suggested that certain lactobacilli might induce dental caries in hyposalivated rats fed a sucrose diet. Three strains of Lactobacillus species isolated from the hyposalivated rats were made resistant to erythromycin. The caries-inducing activity of these erythromycin-resistant lactobacilli was studied in hyposalivated rats giving erythromycin in the drinking water at a concentration of 500 micrograms/ml. After a 61-day experimental period, severe dental caries was induced in hyposalivated rats infected with L. fermentum TY1R. On the other hand, low caries incidence was found in hyposalivated rats infected with either L. acidophilus TY7R or L. plantarum TY3R. These results indicate that L. fermentum may be one of causative agents of dental caries in hyposalivated rats fed a sucrose diet.

Animals↗

[Adhesive properties of lactobacilli isolated from the human gastrointestinal tract].

The adhesive and hemagglutinating properties of 34 tribes of Lactobacillus have been examined. 21 tribes came from human saliva and feces. 23.5% of the tribes showed strongly, 50% weakly adhesive properties. The remaining tribes showed average effects. Most of the strongly adhering tribes were isolates from human specimens. In the group of weakly adhering tribes those from collections dominated. 4 of 9 examined species differed in their adhering capacity. These were L. acidophilus, L. salivarius, L. casei, and L. fermentum. Only weakly adhering tribes showed hemagglutination of human erythrocytes. Therefore, there exists obviously, a reversed relation between the adhering and the hemagglutinating properties of Lactobacilli.

Adult↗

[Inhibitory activity of vaginal Lactobacillus bacteria on yeasts causing vulvovaginal candidiasis].

Growing frequency of therapeutical failures of vulvovaginal candidiasis, resulting from resistance of certain species of Candida to imidazole agents, raises interest in the use of probiotics from Lactobacillus genera as prophylaxis. Unfortunately, little is known about inhibitory mechanisms of Lactobacillus on Candida. The aim of this study was to compare the activity of selected Lactobacillus species, representing the physiological vaginal flora, against Candida as well as investigation whether their inhibitory activity against Candida is related strictly to hydrogen peroxide and lactic acid production. 125 strains from vaginal smears of healthy women were classified by making use of phenotypic and genotypic methods. The majority of strains belonged to L. acidophilus: L. acidophilus sensu stricto, L. crispatus, L. gasseri and L. johnsonii as well as L. fermentum and L. plantarum species. Culture supernatants of selected 25 strains representing the isolated species were examined for their inhibitory activity against the growth of Candida albicans and C. glabrata. The results showed that the strongest and the fastest activity against C. albicans was demonstrated by L. delbrueckii strains, producing the largest quantities of hydrogen peroxide. On the other hand, extended activity, demonstrable after 24 hours, was shown by non-H2O2 producing L. plantarum supernatants. Growth of C. glabrata was not inhibited by any of the examined strains of Lactobacillus. Comparison of activity of live active cultures of Lactobacillus strains and their mixtures with this of pure H2O2 and lactic acid has shown that pure chemical compounds were less active than the cultures. This suggests that mixtures of Lactobacillus strains are in cooperation with each other using many different metabolites.

Candida↗

Pathogenic relevance of Lactobacillus: a retrospective review of over 200 cases.

Given that Lactobacillus has been reported to be the causative pathogen in many types of infection despite debate regarding the organism's clinical significance, a literature review was conducted to investigate the treatments and outcomes of Lactobacillus infections reported to date. In this article, the characteristics of over 200 reported cases of Lactobacillus-associated infections are summarized. Lactobacillus was found to be frequently associated with endocarditis and bacteremia. Lactobacillus was also associated with a variety of other infections including, but not limited to, peritonitis, abscesses, and meningitis. The species casei and rhamnosus were the most common. The isolates tended to be most sensitive to erythromycin and clindamycin and most resistant to vancomycin. The species that was most sensitive to vancomycin was acidophilus. The overall mortality rate was nearly 30%. There was a significant association between mortality and polymicrobial infection (P=0.004). In the subset of patients with bacteremia, increased mortality was associated with inadequate treatment (P=0.001) and polymicrobial bacteremia (P=0.044).

Adolescent↗

Immunological relationships among lactic dehydrogenases in the genera Lactobacillus and Leuconostoc.

Antisera were prepared against pure nicotinamide adenine dinucleotide-dependent d-lactic dehydrogenases of Lactobacillus leichmannii, L. jensenii, and L. fermenti. When tested against these three antisera, crude extracts of almost all species of Lactobacillus containing a d-lactic dehydrogenase give cross-reactions. Extensive pairwise comparisons between cross-reacting crude extracts by double diffusion experiments permit the recognition of groups of identical antigenic specificity among the lactic dehydrogenases of the various nomenspecies of Lactobacillus. The same groups are revealed by each of the three antisera. By analyses of spur formation, the groups of identical antigenic specificity can be arranged in order of decreasing similarity to the homologous d-lactic dehydrogenase used as the reference point. From the combined results obtained with the three antisera, a map of the antigenic relationships among the d-lactic dehydrogenases of lactobacilli can be constructed. Microcomplement fixation experiments with two of the three anti-d-lactic dehydrogenases antisera support the conclusions drawn from double diffusion experiments and provide a quantitative estimation of the antigenic relationships among the various d-lactic dehydrogenases. An antiserum was also prepared against the pure l-lactic dehydrogenase of L. acidophilus group III. It cross-reacts with extracts of almost all lactobacilli containing an l-lactic dehydrogenase. With respect to species that contain both d- and l-lactic dehydrogenases, this antiserum reveals the same groups of identical antigenic specificity as do the antisera directed against d-lactic dehydrogenases. Other than the genus Lactobacillus, only extracts of Leuconostoc cross-react with anti-d-lactic dehydrogenase. No extrageneric cross-reactions were obtained with the anti-l-lactic dehydrogenase.

Animals↗

Antagonistic activity of bacteriocin produced by Lactobacillus species from ogi, an indigenous fermented food.

Seven Lactobacillus species each with one or more strains were isolated from various fermented cereal gruel's (ogi). They were identified as L. plantarum (3 strains), L. delbrueckii (1 strain), L. brevis (2 strains), L. reuteri (2 strains), L. casei (1 strain), L. fermentum (1 strain) and L. acidophilus (1 strain). Bacteriocin production was observed in cell-free supernatants of 8 of these strains with L. fermentum, L. delbrueckii and L. reuteri strains (white maize ogi) being negative. The bacteriocin produced by the eight strains inhibited the growth of various target organisms with the inhibition strongly noticed using Enterococcus faecalis as indicator. While catalase treatment, pH changes and heat treatment up to 80 degrees C had no effect on the activity of bacteriocin from these isolates, treatment with trypsin and proteinase K resulted in complete loss of inhibitory activity of the bacteriocins. A reduction in the inhibitory activity of the bacteriocins was also found to occur with increasing concentrations of glucose or peptone in the cultivation medium.

Bacteriocins↗

Viability of vaginal probiotic lactobacilli during refrigerated and frozen storage.

The viability of six different strains of probiotic vaginal Lactobacillus was examined in two different cryoprotective media, during refrigerated versus frozen storage, and using two traditional types of stock cultures for starting the biomass production. Freezing at -20 degrees C and -70 degrees C had much less adverse effect on viability than did storage at 7 degrees C, and the reduction in viability was greater at -20 degrees C than at -70 degrees C. The strains showed variation in the extent of the viability losses during both types of storage. Milk-yeast extract (MYE) was shown to be the more suitable protective medium to maintain viability of the strains during the storage. The vaginal Lactobacillus strains are most stable in MYE at -70 degrees C with only a small decrease of the viability observed under these conditions. The viable cell counts of Lactobacillus paracasei CRL 1251 and CRL 1289, L. crispatus CRL 1266 and L. salivarius CRL 1328 remained around 1 x 10(8) CFU/mL after 24 months of storage at -70 degrees C, or up to 18 months for L. acidophilus CRL 1259.

Journal Article↗

Genetic diversity, dynamics, and activity of Lactobacillus community involved in traditional processing of artisanal Manchego cheese.

A total of 248 strains of predominant lactobacilli isolated during the manufacture and ripening of artisanal Manchego cheeses obtained from two dairies were obtained and the genetic diversity of 197 investigated using random amplified polymorphic DNA (RAPD-PCR). 51 isolates could not be lysed and were therefore not genotyped. Forty-two distinct RAPD patterns, grouped in six major clusters at a similarity level of 54%, were obtained. Phenotypic characterization of isolates enabled their assignment to the species L. plantarum, L. brevis, L. paracasei subsp. paracasei, L. fermentum, L. pentosus, L. acidophilus and L. curvatus. In samples from both dairies, the species L. plantarum, L. brevis and L. paracasei subsp. paracasei dominated during ripening. Three genotypes showed excellent physiological characteristics and were therefore proposed as adjunct cultures for Manchego cheese manufacture.

Animals↗

Rapid discrimination of lactobacilli isolated from kefir grains by FT-IR spectroscopy.

Fourier transform infrared (FT-IR) spectroscopy was used in combination with multivariate statistical analysis for differentiation of lactic bacteria isolated from kefir grains. Twelve reference strains and 42 lactobacilli isolates from four local kefir grains, previously identified by biochemical traditional techniques at species level were included in this study. The spectra were analysed by hierarchical clustering analysis (HCA) using Pearson's product-moment correlation coefficient and Ward's algorithm. The differentiation between homo- and heterofermentative lactobacilli, proposed as a first level in the classification scheme, was performed with vector normalized first derivatives spectra in the windows 1789-1700, 1059-935, 3000-2927 and 896-833 cm(-1). For heterofermentative lactobacilli the windows 1780-1750, 1500-1200, 2950-2930 and 900-700 cm(-1) were found to contribute to the maximal separation among L. kefir, L. parakefir and Lactobacillus brevis. It was also demonstrated that although this model was robust against small variations in growth temperature (+/-5 degrees C) and growth time (+/-5 h), the make of culture medium used (Biokar or Difco) affected the separation of heterofermentative lactobacilli at species level. For homofermentative lactobacilli the spectral regions 1230-900, 1777-1690, 1357-1240 and 2960-2870 cm(-1), were selected for discrimination among 5 different species that are normally present in kefir grains: L. plantarum, L. acidophilus, L. kefirgranum, L. kefiranofaciens and L. cassei. The classification and discrimination schemes proposed in this work completely matched with the identification obtained by classical biochemical techniques at species level.

Cluster Analysis↗

Assessment of safety of lactobacillus strains based on resistance to host innate defense mechanisms.

Seven Lactobacillus strains belonging to four species were evaluated for pathogenicity as well as for in vitro sensitivity to the bactericidal mechanisms of macrophages in a rabbit infective endocarditis (IE) model. Two bacteremia-associated strains, L. rhamnosus PHLS A103/70 and L. casei PHLS A357/84, as well as the L. rhamnosus type strain and the probiotic L. rhamnosus strain ATCC 53103, showed moderate infectivity, and the virulence of the probiotic L. casei strain Shirota and type strains such as L. acidophilus ATCC 4356(T) and L. gasseri DSM 20243(T) in the model was negligible. The strains that showed pathogenic potential in the rabbit IE model (PHLS A357/84, PHLS A103/70, and ATCC 53103) were more resistant than strain Shirota to intracellular killing activity by mouse macrophages in vitro and also to bactericidal nitrogen intermediates, such as nitric oxide and NO(2)(-) ions. These results suggest that resistance to host innate defense systems, which would function at inflammatory lesions, should be considered in the safety assessment of Lactobacillus strains.

Animals↗

Binding of extracellular matrix proteins by lactobacilli.

Ten gut and ten vaginal Lactobacillus strains were investigated for their ability to bind type I collagen (Cn-I) and four selected gut lactobacilli were investigated for their binding to other extracellular matrix (ECM) molecules. Immobilized Cn-I (100 mg/L) in wells of microtitre plates was bound by all 10 autoaggregating vaginal strains and by 3 strains of gut lactobacilli from piglets in the range of A570 readings 0.114-1.806. L. acidophilus strain SV31 was much more adherent than the rest of strains. All four gut lactobacilli tested for binding to other ECM molecules displayed good binding to porcine fibronectin and heparin and some of them bound weakly to fetuin and porcine mucin. No binding of these strains was observed to bovine mucin, bovine fibrinogen and bovine lactoferrin.

Animals↗

Lactobacillus species identification, H2O2 production, and antibiotic resistance and correlation with human clinical status.

Lactobacilli recovered from the blood, cerebrospinal fluid, respiratory tract, and gut of 20 hospitalized immunocompromised septic patients were analyzed. Biochemical carbohydrate fermentation and total soluble cell protein profiles were used to identify the species. Hydrogen peroxide production was measured. Susceptibility to 19 antibiotics was tested by a diffusion method, and the MICs of benzylpenicillin, amoxicillin, imipenem, erythromycin, vancomycin, gentamicin, and levofloxacin were determined. A small number of species produced H2O2, and antibiotic susceptibilities were species related. Eighteen (90%) of the isolates were L. rhamnosus, one was L. paracasei subsp. paracasei, and one was L. crispatus. L. rhamnosus, L. paracasei subsp. paracasei isolates, and the type strains were neither H2O2 producers nor vancomycin susceptible (MICs, >/=256 microgram/ml). L. crispatus, as well as most of the type strains of lactobacilli which belong to the L. acidophilus group, was an H2O2 producer and vancomycin susceptible (MICs, <4 microgram/ml).

Adolescent↗