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Genomic diversity of cultivable Lactobacillus populations residing in the neonatal and adult gastrointestinal tract.

The objective of this study was to investigate the cultivable Lactobacillus population in adult and infant faecal material to identify strains shared across a number of individuals. A range of lactobacilli isolated on Lactobacillus-selective agar from faeces of 16 infants and 11 adults were genetically fingerprinted and further characterized by 16S rRNA gene sequencing. The relatedness of all the Lactobacillus strains isolated to known species was also determined both genetically and phenotypically. This study revealed that the human intestine is initially colonized by only a few (1-2) different cultivable strains whereas in adults the pattern becomes more complex with a higher diversity of strains. The adult samples contained three genetically distinct Lactobacillus strains in some cases, while infant samples generally harboured only one dominant Lactobacillus strain. Moreover, the species in general appeared to differ with Lactobacillus rhamnosus and Lactobacillus casei/paracasei found mainly in adults, whereas Lactobacillus gasseri and Lactobacillus salivarius were more commonly isolated in infant samples. The data reaffirm the differences in Lactobacillus populations both between individual subjects and between the infant and adult, with an overall change in the diversity and complexity from early stages of life to adulthood.

Adolescent↗

Protective effect of selenium-enriched Lactobacillus on CCl4-induced liver injury in mice and its possible mechanisms.

AIM: To study the protective effects and mechanisms of Se-enriched lactobacillus on liver injury caused by carbon tetrachloride (CCl4) in mice. METHODS: Seventy-two ICR mice were randomly divided into four groups: normal group, CCl4-induced model group, low Se-enriched lactobacillus treatment group (L-Se group), and high Se-enriched lactobacillus treatment group (H-Se group). During a 3-wk experimental period, the common complete diet was orally provided daily for normal group and model group, and the mice in L-Se and H-Se groups were given a diet with 2 and 4 mg of organoselenium from Se-enriched lactobacillus per kg feed, respectively. From the 2nd wk of experiment, the model group, L-Se group, and H-Se group received abdominal cavity injection of olive oil solution containing 500 mL/L CCl4 (0.07 mL/100 g body mass) to induce liver injury, and the normal group was given olive oil on every other day for over 2 wk. In the first 2 wk post injection with CCl4, mice in each group were killed. The specimens of blood, liver tissue, and macrophages in abdominal cavity fluid were taken. Then the activities of the following liver tissue injury-associated enzymes including glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) as well as malondialdehyde (MDA) content were assayed. Changes of phagocytic rate and phagocytic index in macrophages were observed with Wright-Giemsa stain. Plasma TNF-alpha level was measured by radioimmunoassay. The level of intracellular free Ca2+ ([Ca2+]i) in hepatocytes was detected under a laser scanning confocal microscope. RESULTS: During the entire experimental period, the AST and ALT activities in liver were greatly enhanced by CCl4 and completely blunted by both low and high doses of Se-enriched lactobacillus. The Se-enriched lactobacillus-protected liver homogenate GSH-Px and SOD activities were higher or significantly higher than those in model group and were close to those in normal group. CCl4 significantly increased MDA content in liver homogenates, while administration of Se-enriched lactobacillus prevented MDA elevation. Phagocytic rate and phagocytic index of macro-phages decreased after CCl4 treatment compared to those in normal control, but they were dramatically rescued by Se-enriched lactobacillus, showing a greatly higher phagocytic function compared to model group. CCl4 could significantly elevate plasma TNF-alpha and hepatocyte [Ca2+]i level, which were also obviously prevented by Se-enriched lactobacillus. CONCLUSION: Se-enriched lactobacillus can intervene in CCl4-induced liver injury in mice by enhancing macrophage function activity to keep normal and beneficial effects, elevating antioxidant-enzyme activities and reducing lipid peroxidation reaction, inhibiting excessive release of TNF-alpha, preventing the dramatic elevation of [Ca2+]i in hepatocytes.

Alanine Transaminase↗

Rapid identification of probiotic Lactobacillus species by multiplex PCR using species-specific primers based on the region extending from 16S rRNA through 23S rRNA.

This study aimed to develop a novel multiplex polymerase chain reaction (PCR) primer set for the identification of seven probiotic Lactobacillus species such as Lactobacillus acidophilus, Lactobacillus delbrueckii, Lactobacillus casei, Lactobacillus gasseri, Lactobacillus plantarum, Lactobacillus reuteri and Lactobacillus rhamnosus. The primer set, comprising of seven specific and two conserved primers, was derived from the integrated sequences of 16S and 23S rRNA genes and their rRNA intergenic spacer region of each species. It was able to identify the seven target species with 93.6% accuracy, which exceeds that of the general biochemical methods. The phylogenetic analyses, using 16S rDNA sequences of the probiotic isolates, also provided further support that the results from the multiplex PCR assay were trustworthy. Taken together, we suggest that the multiplex primer set is an efficient tool for simple, rapid and reliable identification of seven Lactobacillus species.

Bacterial Typing Techniques↗

Growth phase of orally administered Lactobacillus strains differentially affects IgG1/IgG2a ratio for soluble antigens: implications for vaccine development.

Lactobacillus strains with probiotic activity are major constituents of numerous common food products. Due to their 'generally regarded as safe'-status (GRAS-status), Lactobacillus strains can also be genetically engineered for use in oral immunotherapeutic applications, such as vaccination and T lymphocyte tolerance induction in autoimmune disease.In the current study, we demonstrate that the growth phase of orally administered individual Lactobacillus strains can differentially affect antigen-specific antibody subclasses IgG1 and IgG2a, which might reflect skewing of systemic activity of T helper cell type 2 (Th2) and T helper cell type 1 (Th1) pathways, respectively. Mice were orally fed different wild type Lactobacillus strains in log phase or stationary phase and immunized intraperitoneally with a T-cell dependent protein antigen. Sera were evaluated for the ratio of antigen-specific IgG1 and IgG2a antibodies. Stationary Lactobacillus murines and Lactobacillus casei cultures, but not two other Lactobacillus strains, evoked significantly higher IgG1/IgG2a ratios than log phase cultures, possibly relating to increased activity of the Th2-pathway. Despite normal variation in antibody responses against TNP-CGG among individual mice, a high correlation was found between the IgG1 and IgG2a responses of mice within experimental groups. This differential antibody response is likely due to growth phase-dependent differences in bacterial cell composition.Since Lactobacillus growth phase dependent skewing of antibody responses possibly reflecting T-cell pathways can inadvertently affect allergic and (auto)-immune responses, the current findings strongly caution against unidimensional views on the oral administration of individual Lactobacillus strains for probiotic or immunotherapeutic purposes, but also suggest additional possibilities for immune modulation.

Administration, Oral↗

Design and evaluation of a Lactobacillus group-specific ribosomal RNA-targeted hybridization probe and its application to the study of intestinal microecology in pigs.

The potential importance of anaerobic bacteria belonging to the Lactobacillus group has been well documented. Appropriate methods to rapidly evaluate species diversity and fluctuations in their population levels within the Lactobacillus group are being developed. Molecular tools such as hybridization probes based on rRNA sequences are well suited to these studies. The work reported here was undertaken to test the specificity of an hybridization probe to specifically recognize microorganisms of the Lactobacillus group and assess its usefulness as a quantitative tool to study fluctuations of the Lactobacillus population relative to the total bacterial population in gastrointestinal contents of pigs. We have designed a 25-mer oligonucleotide that targets a region common to and specific for the Lactobacillus group 16S rRNA sequences within the available database. The optimal wash temperature of the probe was experimentally determined to be 54 degrees C. The results obtained using the Lactobacillus group-specific probe (LGP) shows that Lactobacillus populations vary along the different segments of the gastrointestinal tract (GIT). In weaning piglets, the relative Lactobacillus signal intensity obtained constituted 100% of the relative RNA index in the stomach contents as determined by a bacterial domain probe (BDP), and between 90 to 100% in the duodenum. The signal of the Lactobacillus population decreased and reached its minimum in the distal part of the GIT. The same trend was observed in adult pigs, but in the stomach they constituted no more than 30% as determined by the BDP, and were present at lower levels in the other parts of the GIT. These studies document the quantitative importance of the lactobacilli in the stomach and small intestine of pigs. Further studies to investigate the role of lactobacilli in promoting the ecological balance of gut bacteria for probiotic therapy are being undertaken.

Animals↗

Identification of lactobacilli from sourdough and description of Lactobacillus pontis sp. nov.

The microflora of sourdough preparations was investigated by examining the physiological characteristics, whole-cell protein patterns, and 16S rRNA sequences of Lactobacillus isolates. Strains isolated from sourdough were placed in the species Lactobacillus brevis, Lactobacillus sanfrancisco, and Lactobacillus reuteri. 16S rRNA sequences were determined for L. brevis, Lactobacillus fructivorans, Lactobacillus fermentum, L. sanfrancisco, and L. reuteri, and oligonucleotide probes for fast specific identification of these sourdough lactobacilli were deduced. The physiological characteristics, protein patterns, and 16S rRNA sequences of these organisms were compared with data for other sourdough lactobacilli and additional reference strains. Strains of a Lactobacillus species were isolated from rye sourdough; these strains may account for most of the flora in sourdough made from wheat or rye. These organisms were differentiated from other sourdough lactobacilli by their protein pattern, 16S rRNA sequence, G + C content, and physiological properties. The 16S rRNA sequence of this species was determined, and we constructed a phylogenetic tree which reflected the relationships of this species to other lactobacilli. This organism is closely related to L. reuteri. A new Lactobacillus species, Lactobacillus pontis, is proposed. The type strain is L. pontis LTH 2587 (= DSM 8475 = LMG 14187). We describe a general strategy in which a polyphasic approach was used to characterize a new species.

Bacterial Proteins↗

Characterization of lactobacilli by Southern-type hybridization with a Lactobacillus plantarum pyrDFE probe.

Lactobacillus plantarum, Lactobacillus pentosus, and Lactobacillus paraplantarum (M.-C. Curk, J.-C. Hubert, and F. Bringel, Int. J. Syst. Bacteriol. 46:595-598, 1996) can hardly be distinguished on the basis of their phenotypes. Unlike L. plantarum and L. paraplantarum, L. pentosus ferments glycerol and xylose but not melezitose. We identified two L. pentosus strains (CNRZ 1538 and CNRZ 1544) which ferment glycerol and melezitose but not xylose. alpha-Methyl-D-mannoside was fermented by 66% of the L. plantarum strains tested but not by L. paraplantarum strains. In this paper we describe a simple method to identify L. plantarum, L. pentosus, and L. paraplantarum. This method is based on nonradioactive Southern-type hybridization between BglI DNA digests of the lactobacilli tested and a DNA probe (L. plantarum pyrDFE genes from strain CCM 1904). A total of 68 lactobacilli were classified into five groups on the basis of the bands detected. Two groups contained L. plantarum strains; one of these groups contained 31 strains, including the type strain, and was characterized by bands at 7, 4, and 1 kb, and the other group contained strain LP 85-2 and was characterized by bands at 5 and 1.1 kb. Only one band (a band at around 7 kb) was detected in the strains belonging to the L. pentosus group, and two bands (at 4 and 1 kb) were found in the strains belonging to the L. paraplantarum group. No hybridization was detected in the last group, which contained Lactobacillus casei, Lactobacillus coryniformis, Lactobacillus paracasei, Lactobacillus brevis, Lactobacillus delbrueckii, and Lactobacillus leichmannii strains.

Base Sequence↗

Lactobacillus hammesii sp. nov., isolated from French sourdough.

Twenty morphologically different strains were chosen from French wheat sourdough isolates. Cells were Gram-positive, non-spore-forming, non-motile rods. The isolates were identified using amplified-fragment length polymorphism, randomly amplified polymorphic DNA and 16S rRNA gene sequence analysis. All isolates were members of the genus Lactobacillus. They were identified as representing Lactobacillus plantarum, Lactobacillus paralimentarius, Lactobacillus sanfranciscensis, Lactobacillus spicheri and Lactobacillus sakei. However, two isolates (LP38(T) and LP39) could be clearly discriminated from recognized Lactobacillus species on the basis of genotyping methods. 16S rRNA gene sequence similarity and DNA-DNA relatedness data indicate that the two strains belong to a novel Lactobacillus species, for which the name Lactobacillus hammesii is proposed. The type strain is LP38(T) (=DSM 16381(T)=CIP 108387(T)=TMW 1.1236(T)).

Bread↗

Prevention and treatment of vulvovaginal candidiasis using exogenous Lactobacillus.

OBJECTIVE: To review literature examining exogenous Lactobacillus therapy for vulvovaginal candidiasis and to discuss recommendations for clinical practice and future research. DATA SOURCES: Computerized searches on MEDLINE and CINAHL November 2000, September 2001, and March 2002, with search terms including Lactobacillus, acidophilus, Candida, and yeast infections. STUDY SELECTION: Relevant English-language articles from the past 10 years. Unique or seminal studies included where pertinent. DATA EXTRACTION AND SYNTHESIS: Data organized under the following headings: endogenous Lactobacillus, exogenous Lactobacillus, Candida, studies of intravaginal Lactobacillus therapy for vulvovaginal candidiasis, studies of oral Lactobacillus therapy for vulvovaginal candidiasis. CONCLUSIONS: Vaginally administered or orally ingested Lactobacillus is able to colonize the vaginal ecosystem. Controlled intervention studies regarding the effect of such colonization on vulvovaginal candidiasis are promising but few. These studies had small numbers of participants, were inconsistent in the form of Lactobacillus used, and reported conflicting results. Further randomized controlled trials involving large numbers of women are imperative. In the meantime, health care providers should discuss potential benefits with affected patients while clarifying the current lack of conclusive evidence. Without further research into currently available sources and brands of Lactobacillus and without governmental regulation of supplements and their contents, however, it is difficult to make recommendations regarding appropriate product choice.

Administration, Intravaginal↗

Effects of various adjuvants on efficacy of a vaccine against Streptococcus bovis and Lactobacillus spp in cattle.

OBJECTIVE: To determine efficacy of vaccines incorporating QuilA, alum, dextran combined with mineral oil, or Freund adjuvant for immunization of feedlot cattle against Streptococcus bovis and Lactobacillus spp. ANIMALS: 24 steers housed under feedlot conditions. PROCEDURE: Steers were randomly assigned to 4 experimental groups and a control group. Animals in experimental groups were inoculated on days 0 and 26 with vaccines containing Freund adjuvant (FCA), QuilA, dextran combined with mineral oil (Dex), or alum as adjuvant. Serum anti-S bovis and anti-Lactobacillus IgG concentrations were measured, along with fecal pH, ruminal fluid pH, and number of S bovis and Lactobacillus spp in ruminal fluid. RESULTS: Throughout the study, serum anti-S bovis and anti-Lactobacillus IgG concentrations for animals in the Dex, QuilA, and alum groups were similar to or significantly higher than concentrations for animals in the FCA group. Serum anti-S bovis and anti-Lactobacillus IgG concentrations were significantly increased on days 26 through 75 in all 4 experimental groups, and there was a linear relationship between anti-S bovis and anti-Lactobacillus IgG concentrations. For animals in the QuilA and Dex groups, mean pH of feces throughout the period of experiment were significantly higher and numbers of S bovis and Lactobacillus spp in ruminal fluid on day 47 were significantly lower than values for control cattle. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that immunization of feedlot steers against S bovis and Lactobacillus spp with vaccines incorporating Freund adjuvant, QuilA, dextran, or alum as an adjuvant effectively induced high, long-lasting serum anti-S bovis and anti-Lactobacillus IgG concentrations. Of the adjuvants tested, dextran may be the most effective.

Adjuvants, Immunologic↗

Human Milk Oligosaccharides Modulate Nitrogen Utilization in Lactobacillus crispatus in a Glucose-Dependent Manner.

The vaginal microbiome's transition to a dysbiotic state increases susceptibility to pathogens like group B Streptococcus. While human milk oligosaccharides are established prebiotics in the neonatal gut, their impact on the vaginal niche remains largely unexplored. This study investigated the effects of pooled human milk oligosaccharides on the growth and physiology of vaginal (Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus iners) and gut-derived (Lactobacillus reuteri, Lactobacillus rhamnosus) commensals. Growth analyses revealed that human milk oligosaccharides significantly and selectively stimulated growth across all vaginal strains tested, whereas gut commensals exhibited variable or inhibited growth. Carbohydrate utilization assays and comparative genomics against Bifidobacterium infantis showed that Lactobacillus crispatus and Lactobacillus reuteri lack the canonical metabolic machinery to catabolize human milk oligosaccharides. Instead, nitrogen utilization assays identified a glucose-dependent pathway where human milk oligosaccharides are associated with the depletion of primary amines and amino acids in Lactobacillus crispatus supernatants. These results suggest that human milk oligosaccharides act as noncatabolic modulators of vaginal lactobacilli. Collectively, these in vitro findings may warrant investigation of human milk oligosaccharides as modulators of vaginal commensal physiology in more complex experimental systems.

Humans↗

Identification of Lactobacillus species of human origin by a commercial kit, API50CHL.

The efficacy of a biochemical kit, API50CHL kit, for identification of intestinal and vaginal lactobacilli from humans was evaluated by comparing with the results of DNA-DNA hybridization assay. The results showed that in total only 52 of the 172 strains (30.2%) tested were identified correctly by the kit at species level. Especially all strains of some species, such as Lactobacillus gasseri and Lactobacillus crispatus, were misidentified as Lactobacillus acidophilus by the kit. However, the kit was found to be reliable for identification of Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus rhamnosus, and Lactobacillus salivarius. This suggests that the exact identification of Lactobacillus isolates from human stool and vaginal specimens by API50CHL kit is difficult without the support of modern genotypic technique.

Evaluation Studies as Topic↗

Lactobacillus bacteremia: description of the clinical course in adult patients without endocarditis.

Lactobacillus bacteremia in the absence of endocarditis is a rare entity, and the clinical relevance of such bacteremia remains unclear. The clinical courses of lactobacillus bacteremia without endocarditis in 43 previously described patients and 12 new patients were reviewed. Bacteremia with Lactobacillus alone occurred in 34 (62%) of the patients, and 12 (22%) of the patients had bacteremia with other organisms, including Lactobacillus. Lactobacillus was isolated from another site in 18 (33%) of these patients. Intravenous catheter infections were not noted in these patients. Underlying conditions included cancer (6 patients), organ transplantation (9), diabetes mellitus (4), and recent surgery (12). Fever occurred in all patients, and eight (15%) of the patients experienced a sepsis syndrome. The mortality rate was 14%; however, only three deaths were attributed soley to lactobacillus sepsis. Lactobacillus bacteremia is an uncommon condition that usually occurs in patients with severe underlying illnesses and is frequently seen as a part of a polymicrobial infection. Blood cultures positive for Lactobacillus represent true infection and not contamination. Although resistance to commonly used antibiotics is common, the mortality rate associated with this bacteremia appears to be low.

Adult↗

Lactobacillus frumenti sp. nov., a new lactic acid bacterium isolated from rye-bran fermentations with a long fermentation period.

Within the framework of the characterization of the microflora of an industrial sourdough fermentation, strains of Lactobacillus amylovorus, Lactobacillus pontis and two other strains were isolated which could not be associated with a valid species. These latter strains were Gram-positive, catalase-negative, non-spore-forming, non-motile rods that could be clearly differentiated from known species by 16S rDNA sequence analysis. For further characterization, the morphological, physiological (sugar fermentation, formation of DL-lactate, hydrolysis of arginine, growth temperature, CO2 production) and chemotaxonomic (G+C content, cell wall composition, SDS-PAGE of whole-cell proteins) properties were determined. Fitting of the complete 16S rDNA sequence into alignments of such sequences, together with the subsequent phylogenetic calculations, allowed the reconstruction of a phylogenetic tree. These data showed that the two strains were phylogenetically related but formed an independent cluster distinct from their closest neighbours, L. pontis, Lactobacillus panis, Lactobacillus oris, Lactobacillus vaginalis and Lactobacillus reuteri. The results of DNA-DNA hybridization experiments indicated that the two isolates represent a new Lactobacillus species, for which the name Lactobacillus frumenti is proposed; the type strain of this species is DSM 13145T (= LMG 19473T).

Bread↗

Lactobacillus sobrius sp. nov., abundant in the intestine of weaning piglets.

To obtain porcine isolates related to Lactobacillus amylovorus, we screened strains from piglet intestine grown on Lactobacillus-specific MRS agar for hybridization to a fluorescent 16S rRNA-targeted DNA probe. Six strains were isolated and further characterized by phenotypic and molecular taxonomic methods. The isolates were Gram-positive, catalase-negative, facultatively anaerobic rods. They had similar phenotypic characteristics and displayed genomic DNA-DNA relatedness values of >78 % to each other, indicating that they belong to a single species. Comparative 16S rRNA gene sequence analysis demonstrated that the novel isolates were members of Lactobacillus rRNA group I, which includes Lactobacillus delbrueckii, the type species of the genus. Based on 16S rRNA gene sequence similarity, Lactobacillus kitasatonis (99 %), Lactobacillus crispatus (98 %) and Lactobacillus amylovorus (97 %) were the nearest relatives of the novel isolates, but their DNA-DNA relatedness was found to be lower than 49 %. One of the isolates, strain OTU171-001T, was further characterized using physiological and biochemical tests. Together, the results enabled genotypic and phenotypic differentiation of strain OTU171-001T from the other species that showed 16S rRNA gene sequence similarity values greater than 97 %. Strain OTU171-001T merits species status and the name Lactobacillus sobrius sp. nov. is proposed. The type strain is OTU171-001T (= DSM 16698T = NCCB 100067T).

Animals↗

Effect of Lactobacillus isolates on the adhesion of pathogens to chicken intestinal mucus in vitro.

AIMS: The aims of this study were to investigate in vitro the effects of Lactobacillus isolates from a chicken on adhesion of pathogenic Salmonella and Escherichia coli to chicken intestinal mucus obtained from different intestinal regions. METHODS AND RESULTS: Bacteria were labelled by using methyl-1,2-[(3)H]-thymidine. The bacterial adhesion was assessed by measuring the radioactivity of bacteria adhered to the mucus. The results showed that the abilities of Lactobacillus spp. to bind to the same intestinal mucus were higher than those of pathogenic Salmonella and E. coli. Pretreatment of intestinal mucus with Lactobacillus fermentum and Lactobacillus acidophilus, alone or in combination, reduced the adhesion of the tested pathogens, but the reductive extent of pathogenic adhesion by Lactobacillus spp. in combination was relatively high. CONCLUSIONS: The tested bacteria had different adhesions to mucus glycoproteins isolated from different intestinal regions of chicken. Lactobacillus acidophilus and Lact. fermentum in combination revealed a better ability to inhibit attachments of Salmonella and E. coli to chicken intestinal mucus than Lactobacillus sp. alone. SIGNIFICANCE AND IMPACT OF THE STUDY: A mixture of intestinal Lactobacillus spp. from a chicken may play a protective role in excluding pathogenic Salmonella and E. coli from the intestine of chicken.

Animals↗

Comparison of agar plate and real-time PCR on enumeration of Lactobacillus, Clostridium perfringens and total anaerobic bacteria in dog faeces.

AIMS: To compare agar plate and real-time PCR methods on enumeration of total anaerobic bacteria, Lactobacillus and Clostridium perfringens in dog faeces. METHODS AND RESULTS: Thirty-two faecal specimens from Labrador retriever dogs were used to compare agar plate and real-time PCR enumeration methods for Lactobacillus, C. perfringens and total anaerobic bacteria. Total anaerobic bacteria, C. perfringens and Lactobacillus of faeces were counted (as CFU g(-1) faeces) for 48-h incubation at 37 degrees C in an anaerobic gas chamber on genus-selective media. Total genomic DNA from samples was extracted by the QIAamp DNA stool mini kit. The quantification of DNA (as DNA copy per gram faeces) by real-time PCR was performed with a LightCycler system with the QuantiTect SYBR green PCR kit for PCR amplification. The results indicated that there was a significant correlation between CFU and DNA copy of Lactobacillus (R2 = 0.78, P < 0.01) and total anaerobic bacteria (R2 = 0.21, P < 0.05); but no correlation was found between CFU and DNA copy of C. perfringens. The regression equations for Lactobacillus and total anaerobic bacteria were log(DNA copy) = 0.83 x log(CFU) + 1.43 and log(DNA copy) = 1.62 x log(CFU) - 6.32 respectively. CONCLUSIONS: The real-time PCR method could be used to enumerate Lactobacillus within 2 days when compared with plating method which requires 5-6 days. SIGNIFICANCE AND IMPACT OF THE STUDY: The real-time PCR method and the primer set for Lactobacillus spp. harboured in the dog intestine can be used for rapid enumeration of lactobacilli and monitoring of the faecal Lactobacillus community.

Animals↗

Development and evaluation of a suppository formulation containing Lactobacillus and its application in vaginal diseases.

Lactobacillus has long been considered the protective flora in the vagina that displaces and kills vaginal pathogens. Lactic acid, H2O2, and antibacterial agents such as lactocin and bacitracin produced by Lactobacillus act against the vaginal pathogens. The first objective of this research was to develop a local application pharmaceutical formulation of a vaginal suppository containing lyophilized culture of Lactobacillus. The second objective was to establish its in vivo performance by developing in vitro methods of evaluation. Lyophilized culture of Lactobacillus sporogenes was selected for this study. Three formulations of the suppositories were prepared by the molding method. Formulations I, II, and III contained cocoa butter, glycerinated gelatin, and PEG 1000 base, respectively. The prepared suppositories were characterized for physical properties. Assembly to simulate the application site was designed. Methods to evaluate the viability, production of lactic acid, and H2O2 produced by the released Lactobacillus at the application site were developed and the antagonistic activity was demonstrated. From the physical characteristics of the suppository formulations, the glycerinated gelatin suppository (formulation II) containing lyophilized Lactobacillus was found to be satisfactory. The developed assembly was satisfactory in simulating the application site. The Lactobacillus released was viable and exhibited the production of lactic acid, hydrogen peroxide, and antagonistic activity against the uropathogen. The suppository formulation containing Lactobacillus and the methods of its evaluation were successfully developed in this research work and have several applications in the vaginal diseases of women.

Biomass↗