Presence Of Bifidobacteria Research Articles

The Effect of Probiotic Biscuit on Faecal Microbiota in Malnourished Children

A human intervention study on the effect of 90 days supplementation of creamed biscuit with the potential of containing probiotic strain E. faecium IS-27526 at a dose of 108cfu/day on Bifidobacterium in the faecal microbiota of malnourished children under 5 years was conducted. The study was a Randomized Double Blind Placebo Controlled Trial for children under five years old. There were 5 groups with 6 children each: P0 = control biscuit with control cream; P1 = fish and soy protein isolate biscuit with control cream; P2 = control biscuit with probiotic cream; P3 = fish and soy protein isolate biscuit with probiotic cream (every day) and P4 = fish and soy protein isolate biscuit with probiotic cream (every two days). PCR was used to detect Bifidobacterium and Enterococcus faecium in faeces samples. It was concluded after 90 days of supplementation, only 25% of children in the controlled group (P0) harboured Bifidobacterium. In treated groups P1 and P4, the percentage of children positive for Bifidobacterium was approximately 60%, while in groups P2 and P3 the prevalence was 100%. However, the increase of 10% was only demonstrated by P3 as 100% of Bifidobacterium content has been shown in group P2 at the pre-treatment condition. In the controlled group (P0), E. faecium could not be identified either pre or post-intervention, in contrast to the treated groups which showed an increase in some treatments with the highest of 66.7% (group P3). No adverse effect was observed during 90 days of supplementation to children. In conclusion, supplementation of fish and soy protein isolate biscuit and/or probiotic cream every day at a dose of 108cfu/day is considered safe for children under five years old and even increased the number of samples with the presence of bifidobacteria and E. faecium in the fecal microbiota.

Administration of Bovine Milk Oligosaccharide to Weaning Gnotobiotic Mice Inoculated with a Simplified Infant Type Microbiota.

Bovine milk oligosaccharides (BMO) share structural similarity to selected human milk oligosaccharides, which are natural prebiotics for infants. Thus, there is a potential in including BMOs as a prebiotic in infant formula. To examine the in vivo effect of BMO-supplementation on the infant gut microbiota, a BMO-rich diet (2% w/w) was fed to gnotobiotic mice (n = 11) inoculated with an infant type co-culture and compared with gnotobiotic mice receiving a control diet (n = 9). Nuclear magnetic resonance metabolomics in combination with high-throughput 16S rRNA gene amplicon sequencing was used to compare metabolic activity and microbiota composition in different compartments of the lower gastrointestinal tract. BMO components were detected in cecum and colon contents, revealing that BMO was available for the gut bacteria. The gut microbiota was dominated by Enterobacteriaceae and minor abundance of Lactobacilliaceae, while colonization of Bifidobacteriaceae did not succeed. Apart from a lower E. coli population in cecum content and lower formate (in colon) and succinate (in colon and cecum) concentrations, BMO supplementation did not result in significant changes in microbiota composition nor metabolic activity. The present study corroborates the importance of the presence of bifidobacteria for obtaining microbial-derived effects of milk oligosaccharides in the gastrointestinal tract.

INTESTINAL MICROFLORA ON THE BACKGROUND OF BACTERIAL VAGINOSIS ON VULVOVAGINAL CANDIDIASIS IN OVERWEIGHT AND OBESE WOMEN

The aim of our work was to study the intestinal microbiome in obese women and further develop differentiated patterns of exposure to the intestinal microbiota to improve metabolism and reduce excess weight. Materials and methods: Surveyed 120 overweight and obese women. For comparison, 60 women without overweight and obesity were examined. Group I – women with vulvovaginal candidiasis and overweight and obesity (60 women); group II – women with vulvovaginal candidiasis without overweight and obesity (30 women); group III – women with bacterial vaginosis and overweight and obesity (60 women); ІV – women with bacterial vaginosis without overweight and obesity (30 women). The study of the intestinal microbiocenosis included the determination of the species and quantitative composition of the microflora. Quantitative indicators of intestinal microflora were studied by seeding 1 ml from each dilution on differential diagnostic media: Endo, Ploskireva, ICA (bismuth – agar sulfide) to detect pathogenic enterobacteria; ZhSA (yellow – salt agar) for determination of staphylococci. To study the hemolytic activity of bacteria was used agar with 5% erythrocyte content of sheep. The presence of bifidobacteria in the test material was studied on Blaurok medium, and lactobacilli – on MRS medium. Statistical analysis of the obtained research results was performed using standard computer packages “Data Analysis” Microsoft Excel for Windows 2002. Results: Based on the results of studies, we can summarize the nature of changes in the intestinal microbiome on the background of vulvovaginal candidiasis and bacterial vaginosis in women with overweight and obesity. In all women with excess body weight and obesity, intestinal microflora disorders have been registered, which are characterized by the replacing the lactobacilli and bifidum bacteria to opportunistic pathogens. A significant increase in Firmicutes phylum microorganisms and a significant decrease in Bacteroidetes phylum were found. The obtained data prove that changes in the species composition of the intestinal microbiota play an important role in the pathogenesis of obesity. Conclusions: The results indicate the feasibility of monitoring the intestinal microbiota in women with vulvovaginal candidiasis and bacterial vaginosis, especially in the presence of excess body weight and obesity to determine the degree of violations of its components, and timely correction of the detected changes.

Exploring the Ecology of Bifidobacteria and Their Genetic Adaptation to the Mammalian Gut.

The mammalian gut is densely inhabited by microorganisms that have coevolved with their host. Amongst these latter microorganisms, bifidobacteria represent a key model to study host–microbe interaction within the mammalian gut. Remarkably, bifidobacteria naturally occur in a range of ecological niches that are either directly or indirectly connected to the animal gastrointestinal tract. They constitute one of the dominant bacterial members of the intestinal microbiota and are among the first colonizers of the mammalian gut. Notably, the presence of bifidobacteria in the gut has been associated with several health-promoting activities. In this review, we aim to provide an overview of current knowledge on the genetic diversity and ecology of bifidobacteria. Furthermore, we will discuss how this important group of gut bacteria is able to colonize and survive in the mammalian gut, so as to facilitate host interactions.

The Infant-Derived Bifidobacterium bifidum Strain CNCM I-4319 Strengthens Gut Functionality.

Bifidobacteria are among the first colonisers of the gastrointestinal tract of breast-fed newborns due to, among other things, their ability to metabolise oligosaccharides naturally occurring in human milk. The presence of bifidobacteria in the infant gut has been shown to promote intestinal health and homeostasis as well as to preserve a functional gut barrier, thus positively influencing host health and well-being. Among human-associated gut commensals, Bifidobacterium bifidum has been described as the only species capable of the extracellular degradation of both mucin-type glycans and HMOs, thereby giving this species a special role as a commensal gut forager of both host and diet-derived glycans. In the present study, we assess the possible beneficial properties and probiotic potential of B. bifidum strain CNCM I-4319. In silico genome analysis and growth experiments confirmed the expected ability of this strain to consume HMOs and mucin. By employing various animal models, we were also able to assess the ability of B. bifidum CNCM I-4319 to preserve gut integrity and functionality from stress-induced and inflammatory damage, thereby enforcing its potential as an effective probiotic strain.

Evolutionary relationships among bifidobacteria and their hosts and environments

BackgroundThe assembly of animal microbiomes is influenced by multiple environmental factors and host genetics, although the relative importance of these factors remains unclear. Bifidobacteria (genus Bifidobacterium, phylum Actinobacteria) are common first colonizers of gut microbiomes in humans and inhabit other mammals, social insects, food, and sewages. In humans, the presence of bifidobacteria in the gut has been correlated with health-promoting benefits. Here, we compared the genome sequences of a subset of the over 400 Bifidobacterium strains publicly available to investigate the adaptation of bifidobacteria diversity. We tested 1) whether bifidobacteria show a phylogenetic signal with their isolation sources (hosts and environments) and 2) whether key traits encoded by the bifidobacteria genomes depend on the host or environment from which they were isolated. We analyzed Bifidobacterium genomes available in the PATRIC and NCBI repositories and identified the hosts and/or environment from which they were isolated. A multilocus phylogenetic analysis was conducted to compare the genetic relatedness the strains harbored by different hosts and environments. Furthermore, we examined differences in genomic traits and genes related to amino acid biosynthesis and degradation of carbohydrates.ResultsWe found that bifidobacteria diversity appears to have evolved with their hosts as strains isolated from the same host were non-randomly associated with their phylogenetic relatedness. Moreover, bifidobacteria isolated from different sources displayed differences in genomic traits such as genome size and accessory gene composition and on particular traits related to amino acid production and degradation of carbohydrates. In contrast, when analyzing diversity within human-derived bifidobacteria, we observed no phylogenetic signal or differences on specific traits (amino acid biosynthesis genes and CAZymes).ConclusionsOverall, our study shows that bifidobacteria diversity is strongly adapted to specific hosts and environments and that several genomic traits were associated with their isolation sources. However, this signal is not observed in human-derived strains alone. Looking into the genomic signatures of bifidobacteria strains in different environments can give insights into how this bacterial group adapts to their environment and what types of traits are important for these adaptations.

Litter feed additive as source of amino acids and beneficial bacteria

Feed additive was derived from poultry manure by microbiological synthesis. After a 10-day feeding poultry with probiotic strains of bacteria B. subtilis TNP-3 and B. subtilis TNP-5, the litter does not contain potential enteropathogens and can be used as a raw material for feed additive. Based on the results of microbiological and biochemical studies the technology of feed additive (powder) with the use of extrusion was developed. Extrusion at a temperature of up to 120 °C for 5-6 seconds provides presence of beneficial bacteria and significantly high content of essential amino acids. According to the results of biochemical studies, litter feed additive contains 18 free amino acids. The total concentration of free amino acids in the feed additive (powder) is 406.3 mg/kg, which is 1.7 times higher than that in the litter without fermentation and extrusion. The experiments have shown that inclusion of 3.3% feed additive in the diet does not have negative effect on physiological state, viability and productivity of laying hens. Survival of birds in both groups was 100%. Additive application in the experimental group of chickens revealed absence of opportunistic pathogenic microorganisms and microscopic fungi, presence of bifidobacteria and spore-forming aerobic Bacillus bacteria in powder, as well as predominance of beneficial micro flora and lack of potential enteropathogens (compared to control). It allows to conclude that feed additive obtained by microbiological synthesis from bird droppings possess probiotic properties. The results of biochemical study of egg production indicate that the use of food additives (to 3.3% of the basic diet) for laying hens significantly increases content of major micro and macro-elements in eggs, compared to the control. Therefore, the use of feed additive-powder (up to 3.3% of the basic diet) does not reduce egg quality. Thus, based on the results of these studies, it can be concluded that the litter obtained from laying hens, after application of probiotic ‘Nord-Bakt’, further fermentation with strains Bacillus subtilis TNP-3 and Bacillus subtilis TNP-5, followed by extrusion can be used as a feed additive as a source of amino acids and beneficial bacteria.

Metabolism of Milk Oligosaccharides in Preterm Pigs Sensitive to Necrotizing Enterocolitis.

Human milk oligosaccharides (HMO) are major components of breast milk that may have local effects in the gastrointestinal tract and systemic functions after being absorbed, both depending on their metabolism. Using preterm pigs, we investigated the metabolic fate of HMO in three experiments with two different HMO blends. In addition, we examined effects on the colonic microbiota in the presence or absence of necrotizing enterocolitis (NEC). Thus, preterm pigs (n = 112) were fed formula without or with HMO supplementation (5–10) g/L of a mixture of 4 (4-HMO) or >25 HMO (25-HMO) for 5 (Experiment 1 and 2) or 11 days (Experiment 3). Individual HMO were quantified in colon contents and urine using MALDI-TOF-MS (matrix-assisted laser desorption ionization mass spectrometry) and HPAEC-PAD (high-performance anion-exchange chromatography with pulsed amperometric detection). Microbial colonization was analyzed by sequencing of 16S rRNA gene tags. Intestinal permeability was measured by lactulose to mannitol ratio in urine. HMO supplemented to formula were detected in urine and colon contents in preterm piglets after 5 and 11 days in all three experiments. The amount of HMO excreted via the gut or the kidneys showed large individual variations. Microbial diversity in the colon changed from high levels of Firmicutes (dominated by Clostridium) at day 5 (Exp 2) to high levels of Proteobacteria dominated by Helicobacter and Campylobacter at day 11 (Exp 3). Colonic microbiota composition as well as HMO excretion pattern varied greatly among piglets. Interestingly, the 5-day supplementation of the complex 25-HMO blend led to low concentrations of 3-fucosyllactose (FL) and lacto-N-fucopentaose (LNFP) I in colonic contents, indicating a preferred utilization of these two HMO. Although the interpretation of the data from our piglet study is difficult due to the large individual variation, the presence of Bifidobacteria, although low in total numbers, was correlated with total HMO contents, and specifically with 2′FL levels in colonic content. However, early supplementation of formula with HMO did not affect NEC incidence.

Resistant Starch Supplementation Improves Bifidobacteria Colonization and Lipid Profile in Adult Obese Subjects Residing in Urban Vadodara

Obesity continues to be a growing problem globally among all age groups, further associated with life threatening co-morbidities. Resistant Starch (RS) as a dietary component has been recently recognized as a prebiotic with potential role in improving the bacterial ecology of human gut. The present study was undertaken to evaluate the impact of resistant starch supplementation by the obese adults in urban Vadodara on microbial changes in gut flora with respect to Bifidobacteria and Enterobacteria. Using a non randomised control trial study design, 60 obese adults were enrolled from the free living population of urban Vadodara. Baseline information was collected and subjects were classified into control (n=30) and experimental group (n=30). Experimental group was supplemented with 10 g of RS daily for the period of two months (60 days). The control group were not given anything during this period. Blood and stool samples were analysed for lipid profile and the presence of Bifidobacteria and pathogenic Enterobacteria, respectively both at baseline and after the intervention period. Results revealed a significant increase in the mean log counts of Bifidobacteria by 11.7% with no significant reduction in the mean log counts of enteric pathogens and lipid profile of subjects. Hence study concludes that a daily supplementation of 10 grams of RS for 60 days to the obese subjects improved colonization of beneficial gut microbiota in terms of Bifidobacteria, but had little effect on obesity.

Role of microorganisms present in dairy fermented products in health and disease.

Role of microorganisms present in dairy fermented products in health and disease.

Comparison of mupirocin-based media for selective enumeration of bifidobacteria in probiotic supplements

An international standard already exists for the selective enumeration of bifidobacteria in milk products. This standard uses Transgalactosylated oligosaccharides (TOS) propionate agar supplemented with mupirocin. However, no such standard method has been described for the selective enumeration of bifidobacteria in probiotic supplements, where the presence of bifidobacteria is much more variable than in milk products. Therefore, we enumerated bifidobacteria by colony count technique in 13 probiotic supplements using three media supplemented with mupirocin (Mup; 100mg/l): TOS, Bifidobacteria selective medium (BSM) and modified Wilkins-Chalgren anaerobe agar with soya peptone (WSP). Moreover, the potential growth of bifidobacterial strains often used in probiotic products was performed in these media. All 13 products contained members of the genus Bifidobacterium, and tested mupirocin media were found to be fully selective for bifidobacteria. However, the type strain Bifidobacterium bifidum DSM 20456 and collection strain B. bifidum DSM 20239 showed statistically significant lower counts on TOS Mup media, compared to BSM Mup and WSP Mup media. Therefore, the TOS Mup medium recommended by the ISO standard cannot be regarded as a fully selective and suitable medium for the genus Bifidobacterium. In contrast, the BSM Mup and WSP Mup media supported the growth of all bifidobacterial species.

Pseudoscardovia radai sp. nov., a representative of the family Bifidobacteriaceae isolated from the digestive tract of a wild pig (Sus scrofa scrofa)

Presence of bifidobacteria and representatives of the new genus Pseudoscardovia within the family Bifidobacteriaceae in the digestive tract of wild pigs has been reported recently. Results based on comparative 16S rRNA gene sequence analysis of a new fructose-6-phosphate phosphoketolase-positive bacterial isolate originated from the small intestine of a wild pig revealed a relationship to Pseudoscardovia suis DPTE4T (96.8% sequence similarity). Phylogenetic and comparative analyses based on 16S rRNA, hsp60, xfp, fusA, tuf and rpoC partial gene sequences confirmed relationship of the new bacterial strain to Pseudoscardovia suis compared with bifidobacteria species occurring in the digestive tract of domestic and wild pigs. Differences in utilization of various substrates, production of enzymes, cell morphology, peptidoglycan structure, profile of cellular fatty acids and polar lipids between the new bacterial isolate designated as DPVI-TET3T and P. suis DPTE4T allow to establish a new bacterial taxon for which the name Pseudoscardovia radai sp. nov. (= DPVI/TET3T = CCM 7943T = DSM 24742T) was proposed.

Factors involved in the colonization and survival of bifidobacteria in the gastrointestinal tract

Probiotics are live microorganisms that when administered in adequate amounts confer a health benefit on the host. They are mainly bacteria from the genera Lactobacillus and Bifidobacterium. Traditionally, functional properties of lactobacilli have been studied in more detail than those of bifidobacteria. However, many recent studies have clearly revealed that the bifidobacterial population in the human gut is far more abundant than the population of lactobacilli. Although the 'beneficial gut microbiota' still remains to be elucidated, it is generally believed that the presence of bifidobacteria is associated with a healthy status of the host, and scientific evidence supports the benefits attributed to specific Bifidobacterium strains. To carry out their functional activities, bifidobacteria must be able to survive the gastrointestinal tract transit and persist, at least transiently, in the host. This is achieved using stress response mechanisms and adhesion and colonization factors, as well as by taking advantage of specific energy recruitment pathways. This review summarizes the current knowledge of the mechanisms involved in facilitating the establishment, colonization, and survival of bifidobacteria in the human gut.

Inter-species differences in the growth of bifidobacteria cultured on human milk oligosaccharides

Human milk (HM) contains as the third most abundant component around 200 different structures of human milk oligosaccharides (HMOs). HMOs are the first and irreplaceable prebiotics for infants, supporting bifidobacteria as the most important bacterial group in an infant intestine. The aim of our study was to test the growth of bifidobacteria in HM and on HMOs. Bifidobacteria were isolated from two groups of infants. The first one (eight strains) were isolated from infants who had bifidobacteria in their feces but, after a short period of time (4 to 24days), bifidobacteria were no longer detected in their feces (disappeared bifidobacteria [DB]). The second group of bifidobacteria (eight strains) originated from infants with continual presence of bifidobacteria in their feces (persistent bifidobacteria [PB]). There were significant differences (p < 0.05) between DB and PB groups in the ability of the strains to grow in HM. PB grew in HM, reaching counts higher than 7 log CFU/ml. In contrast, counts of DB decreased from 5 to 4.3 log CFU/ml after cultivation in HM. The final pH after cultivation of bifidobacteria on HMOs was 6.2 and 4.9 in DP and PB groups, respectively. In general, Bifidobacterium bifidum and B. breve species were able to utilize HMOs, while B. adolescentis and B. longum subsp. longum species did not. The ability to grow in HM and to utilize HMOs seem to be important properties of bifidobacteria which are able to colonize infant intestinal tract.

The presence of bifidobacteria in social insects, fish and reptiles

The occurrence and species distribution of bifidobacteria in the digestive tract of important representatives of social insects such as ants, bees, wasps and bumblebees as well as the incidence of bifidobacteria in fecal samples of several species of vertebrates represented mainly by reptiles was assigned by culture-independent method based on DGGE and real time PCR. Bifidobacteria were present in the gut of most social insects--honey bees, wasps, cockroaches and bumblebees, except for ants. In honey bees, where the counts of bifidobacteria ranged from 2 to 8% of the total bacteria, the most common species seemed to be Bifidobacterium indicum. Proportion of bifidobacteria was found in broad range from 0.1 to 35-37% in wasps and cockroaches; the variance of bifidobacteria in bumblebees was lower, ranging from 1 to 7% of total bacterial count. Among studied vertebrates, the detectable presence of bifidobacteria was found only in trout (1.1%) and geckos (0.2%), but large amount of these bacteria was observed in Vietnamese box turtle, where bifidobacteria represented nearly one-fourth (22%) of total bacterial counts.

Inhibitory Impact of Bifidobacteria on the Transfer of β-Lactam Resistance among Enterobacteriaceae in the Gnotobiotic Mouse Digestive Tract

While looking for new means to limit the dissemination of antibiotic resistance, we evaluated the role of potentially probiotic bifidobacteria on the transfer of resistance genes between enterobacteria. Transfers of bla genes encoding extended-spectrum beta-lactamases (SHV-5 and CTX-M-15) were studied in the absence or presence of bifidobacteria. In vitro, transfer frequencies of these bla genes decreased significantly in the presence of three of five tested strains, i.e., Bifidobacterium longum CUETM-89-215, Bifidobacterium bifidum CIP-56.7T, and Bifidobacterium pseudocatenulatum CIP-104168T. Four transfer experiments were conducted in the digestive tract of gnotobiotic mice, the first three observing the effect of B. longum CUETM-89-215, B. bifidum CIP-56.7T, and B. pseudocatenulatum CIP-104168T on blaSHV-5 transfer and the fourth experiment studying the effect of B. bifidum CIP-56.7T on blaCTX-M-15 transfer. These experiments revealed significant decreases in the transconjugant levels (up to 3 logs) in mice having received B. bifidum CIP-56.7T or B. pseudocatenulatum CIP-104168T compared to control mice. Bifidobacteria appear to have an inhibitory impact on the transfer of antibiotic resistance genes. The inhibitory effect is associated to specific bifidobacterial strains and may be related to the production of thermostable metabolites by these strains.

Properties of Streptococcus thermophilus fermented milk containing variable concentrations of Bifidobacterium longum and Lactobacillus acidophilus

Sensory evaluation and analysis of pH, titratable acidity and microbial counts after 1, 7, 14 and 21 days of storage of five combinations of lyophilized Bifidobacterium longum and/or Lactobacillus acidophilus added to milk fermented with Streptococcus thermophilus were studied during storage at 4oC. The taste and acidity sensory attributes were significantly (P<0.05) perceived as the storage time increased. Favourite combinations contained, initially, Bif. longum and L. acidophilus (108 and 107 cfu/mL, respectively) and Bif. longum only (108 cfu/mL). These combinations did not differ significantly among them and neither from the standard fermented milk. The lowest sensory scores and highest titratable acidity values were obtained for fermented milk added of L. acidophilus (108 cfu/mL initial concentration). During the 21 days of storage at 4oC the viable cell counts of: Str. thermophilus did not change. Bif. longum viable cells counts kept constant or reduced 1 logarithmic cycle and L. acidophilus reduced 1 or 2 logarithmic cycle. We did not observe inhibition of over-acidification caused by the presence of bifidobacteria or L. acidophilus.

Comparison between Gram stain and culture for the characterization of vaginal microflora: Definition of a distinct grade that resembles grade I microflora and revised categorization of grade I microflora

BackgroundThe microbiological diagnosis of bacterial vaginosis is usually made using Nugent's criteria, a useful but rather laborious scoring system based on counting bacterial cell types on Gram stained slides of vaginal smears. Ison and Hay have simplified the score system to three categories and added a fourth category for microflora with a predominance of the Streptococcus cell type. Because in the Nugent system several cell types are not taken into account for a final score, we carried out a detailed assessment of the composition of the vaginal microflora in relation to standard Gram stain in order the improve the diagnostic value of the Gram stain. To this purpose we compared Gram stain based categorization of vaginal smears with i) species specific PCR for the detection of Gardnerella vaginalis and Atopobium vaginae and with ii) tDNA-PCR for the identification of most cultivable species.ResultsA total of 515 samples were obtained from 197 pregnant women, of which 403 (78.3%) were categorized as grade I microflora, 46 (8.9%) as grade II, 22 (4.3%) as grade III and 8 (1.6%) as grade IV, according to the criteria of Ison and Hay. Another 36 samples (7.0%) were assigned to the new category 'grade I-like', because of the presence of diphtheroid bacilli cell types. We found that 52.7% of the grade I-like samples contained Bifidobacterium spp. while L. crispatus was present in only 2.8% of the samples and G. vaginalis and A. vaginae were virtually absent; in addition, the species diversity of this category was similar to that of grade II specimens.Based on the presence of different Lactobacillus cell types, grade I specimens were further characterized as grade Ia (40.2%), grade Iab (14.9%) and grade Ib (44.9%). We found that this classification was supported by the finding that L. crispatus was cultured from respectively 87.0% and 76.7% of grade Ia and Iab specimens while this species was present in only 13.3% of grade Ib specimens, a category in which L. gasseri and L. iners were predominant.ConclusionFurther refinement of Gram stain based grading of vaginal smears is possible by distinguishing additional classes within grade I smears (Ia, Iab and Ib) and by adding a separate category, designated grade I-like. A strong correlation was found between grade Ia and the presence of L. crispatus and between grade I-like and the presence of bifidobacteria. This refinement of Gram stain based scoring of vaginal smears may be helpful to improve the interpretation of the clinical data in future studies, such as the understanding of response to treatment and recurrence of bacterial vaginosis in some women, and the relationship between bacterial vaginosis and preterm birth.

Discrimination between Bifidobacterium Species from Human and Animal Origin by PCR–Restriction Fragment Length Polymorphism

Bifidobacteria are normal intestinal flora in humans and animals. The genus Bifidobacterium includes 31 species of significant host specificity. Taking into account their properties, we proposed to use bifidobacteria as fecal contamination indicators. PCR–restriction fragment length polymorphism on the 16S rDNA gene was used to distinguish the different Bifidobacterium species. Sixty-four strains belonging to 13 different species were differentiated from animal or human origin using one or two restriction enzymes. Moreover, the primers used were specifics of the Bifidobacterium genus. Therefore, this method made it possible to determine both the presence of bifidobacteria in a sample and its origin of contamination.

Viscosity and Proteolysis of Yoghurt Using Ropy Strains, in the Presence of Bifidobacteria, as Compared to Those of Soyoghurt

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