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The flora of the healthy vagina is dominated byH2O2-producing Lactobacillus species, predominantly Lactobacillus crispatus and Lactobacillus jensenii [1]. Bacterial vaginosis (BV) is characterized by overgrowth of commensal anaerobic flora relative to lactobacilli. BV among heterosexual women is associated with a new male sex partner and unprotected intercourse [2, 3] and with vaginal intercourse immediately after anal intercourse [4]. The prevalence of BV among lesbians is high relative to that among heterosexual women [5, 6], and BV is frequently found in both members of lesbian couples [5]. Sexual practices involving digital-vaginal or digital-anal contact might transmit vaginal fluid and may promote abnormal vaginal flora [7]. Criswell et al. [8] “transmitted” BV from one woman to another by transferring vaginal secretions. We hypothesized that sexual behaviors that could transfer vaginal fluid might also transmit lactobacilli between female sex partners. Repetitive element sequence– based PCR (rep-PCR), a DNA fingerprinting technique, has been successful in distinguishing a probiotic Lactobacillus crispatus strain from other L. crispatus strains and from other endogenous Lactobacillus species [9]. Rep-PCR uses repetitive sequences throughout the genome for direct amplification of genomic DNA, generating fingerprint patterns unique to bacterial species or strains [10]. We defined species-specific distribution of genital Lactobacillus isolates recovered from women who reported having had sex with other women and used rep-PCR fingerprinting to assess whether unique Lactobacillus strains were shared by female sex partners.

WOS: 000269654300006, In this work, autoaggregation and factors involved in the autoaggregation ability of vaginal lactobacilli were studied. The autoaggregation ability of 28 lactobacilli strains was positive. The effects of various factors on autoaggregation were also evaluated. Lactobacillus jensenii A1, L. salivarius I1, and L. cellobiosus I3 showed higher autoaggregation in acidic conditions and lower autoaggregation in hot (70 and 85 degrees C), conditions. The L. salivarius I1 strain, which exhibited high autoaggregation activity, also showed good autoaggregation in pepsin, lipase, and sodium periodate, as well as under conditions of sonication and heat. The results of this study suggest that lactobacilli showing high autoaggregation may constitute an important host defense mechanism against infections as a probiotic., Turkish Prime Ministry State Planning Organization [2003 K 12470-06], This study was supported by the Turkish Prime Ministry State Planning Organization (Project No. 2003 K 12470-06). The authors thank Paul Dansted for proofreading the manuscript.

Our objective was to determine the morphotype of the adherent bacteria in liquid-based cytology (LBC) in smears with healthy and disturbed vaginal flora. And to use PCR technology on the same fixed cell sample to establish DNA patterns of the 16S RNA genes of the bacteria in the sample. Thirty samples were randomly selected from a large group of cervical cell samples suspended in a commercial coagulant fixative "(BoonFix)." PCR was used to amplify DNA of five bacterial species: Lactobacillus acidophilus, Lactobacillus crispatus, Lactobacillus jensenii, Gardnerella vaginalis, and Mycoplasma hominis. The LBC slides were then analyzed by light microscopy to estimate bacterial adhesion. DNA of lactobacilli was detected in all cell samples. Seventeen smears showed colonization with Gardnerella vaginalis (range 2.6 x 10(2)-3.0 x 10(5) bacteria/mul BoonFix sample). Two cases were identified as dysbacteriotic with high DNA values for Gardnerella vaginalis and low values for Lactobacillus crispatus. The sample with the highest concentration for Gardnerella vaginalis showed an unequivocal Gardnerella infection. This study indicates that the adherence pattern of a disturbed flora in liquid-based cervical samples can be identified unequivocally, and that these samples are suitable for quantitative PCR analysis. This cultivation independent method reveals a strong inverse relationship between Gardnerella vaginalis and Lactobacillus crispatus in dysbacteriosis and unequivocal Gardnerella infection.

The bacterial biota of the human vagina can have a profound impact on the health of women and their neonates. Changes in the vaginal microbiota have been associated with several adverse health outcomes including premature birth, pelvic inflammatory disease, and acquisition of HIV infection. Cultivation-independent molecular methods have provided new insights regarding bacterial diversity in this important niche, particularly in women with the common condition bacterial vaginosis (BV). PCR methods have shown that women with BV have complex communities of vaginal bacteria that include many fastidious species, particularly from the phyla Bacteroidetes and Actinobacteria. Healthy women are mostly colonized with lactobacilli such asLactobacillus crispatus,Lactobacillus jensenii, andLactobacillus iners, though a variety of other bacteria may be present. The microbiology of BV is heterogeneous. The presence ofGardnerella vaginalisandAtopobium vaginaecoating the vaginal epithelium in some subjects with BV suggests that biofilms may contribute to this condition.

MucoCept is a biotherapeutic for prevention of HIV-1 infection in women and contains a human, vaginal Lactobacillus jensenii that has been genetically enhanced to express the HIV-1 entry inhibitor, modified cyanovirin-N (mCV-N). The objective of this study was to develop a solid vaginal dosage form that supports sustained vaginal colonization of the MucoCept Lactobacillus at levels previously shown, with freshly prepared cultures, to protect macaques from SHIV infection and to test this formulation in a macaque vaginal colonization model. Vaginally disintegrating tablets were prepared by lyophilizing the formulated bacteria in tablet-shaped molds, then packaging in foil pouches with desiccant. Disintegration time, potency and stability of the tablets were assessed. For colonization, non-synchronized macaques were dosed vaginally with either one tablet or five tablets delivered over five days. Vaginal samples were obtained at three, 14, and 21 days post-dosing and cultured to determine Lactobacillus colonization levels. To confirm identity of the MucoCept Lactobacillus strain, genomic DNA was extracted from samples on days 14 and 21 and a strain-specific PCR was performed. Supernatants from bacteria were tested for the presence of the mCV-N protein by Western blot. The tablets were easy to handle, disintegrated within two minutes, potent (5.7x1011 CFU/g), and stable at 4°C and 25°C. Vaginal administration of the tablets to macaques resulted in colonization of the MucoCept Lactobacillus in 66% of macaques at 14 days post-dosing and 83% after 21 days. There was no significant difference in colonization levels for the one or five tablet dosing regimens (p=0.88 Day 14, p=0.99 Day 21). Strain-specific PCR confirmed the presence of the bacteria even in culture-negative macaques. Finally, the presence of mCV-N protein was confirmed by Western blot analysis using a specific anti-mCV-N antibody.

Lactobacilli are Gram-positive anaerobic rods or coccobacilli, commonly found as commensals in human mucosa. Rarely, they can cause serious infections such as infective endocarditis (IE), and the most frequently implicated species causing serious infections are L. casei and L. rhamnosus. IE caused by Lactobacillus jensenii is very rare, with only six reported cases so far, to the best of our knowledge. We present a case of native aortic valve endocarditis caused by L. jensenii, complicated by root abscess and complete heart block, and requiring emergent surgical intervention.

Aims: To investigate the production of antioxidant activity during fermentation with commonly used dairy starter cultures. Moreover, to study the development of antioxidant activity during fermentation, and the connection to proteolysis and bacterial growth. Methods and Results: Antioxidant activity was measured by analysing the radical scavenging activity using a spectrophotometric decolorization assay and lipid peroxidation inhibition was assayed using liposomal model system with a fluorescence method. Milk was fermented with 25 lactic acid bacterial (LAB) strains, and from these six strains, exhibiting the highest radical scavenging activity was selected for further investigation. Leuconostoc mesenteroides ssp. cremoris strains, Lactobacillus jensenii (ATCC 25258) and Lactobacillus acidophilus (ATCC 4356) showed the highest activity with both the methods used. However, the radical scavenging activity was stronger than lipid peroxidation inhibition activity. The development of radical scavenging activity was connected to proteolysis with four strains. Molecular distribution profiles showed that fermentates with high scavenging activity also possessed a higher proportion of peptides in the molecular mass range of 4–20 kDa, while others had mostly large polypeptides and compounds below 4 kDa. In addition, the amount of hydrophobic amino acids was higher in these fermentates. Conclusions: The development of antioxidant activity was strain-specific characteristic. The development of radical scavengers was more connected to the simultaneous development of proteolysis whereas, lipid peroxidation inhibitory activity was related to bacterial growth. However, high radical scavenging activity was not directly connected to the high degree of proteolysis Significance and Impact of the Study: To the best of our knowledge, this seems to be the first report, which screens possible antioxidant activity among most common dairy LAB strains. Use of such strains improve nutritional value of fermented dairy products.

International audience; The purpose of this study was to investigate how human vaginal isolates of Lactobacillus acidophilus, Lactobacillus jensenii, Lactobacillus gasseri and Lactobacillus crispatus inhibit the vaginosis-associated pathogens Gardnerella vaginalis and Prevotella bivia. Results show that all the strains in coculture condition reduced the viability of G. vaginalis and P. bivia, but with differing degrees of efficacy. The treatment of G. vaginalis- and P. bivia-infected cultured human cervix epithelial HeLa cells with L. gasseri strain KS120.1 culture or cell-free culture supernatant (CFCS) results in the killing of the pathogens that are adhering to the cells. The mechanism of the killing activity is not attributable to low pH and the presence of lactic acid alone, but rather to the presence of hydrogen peroxide and proteolytic enzyme-resistant compound(s) present in the CFCSs. In addition, coculture of G. vaginalis or P. bivia with L. gasseri KS120.1 culture or KS120.1 bacteria results in inhibition of the adhesion of the pathogens onto HeLa cells.

Results A human vaginal isolate of Lactobacillus jensenii was engineered, by stable integration of an optimized expression cassette into the bacterial genome, to secrete high levels of the highly potent HIV inhibitor, CV-N. The L. jenseniiexpressed CV-N dramatically decreases infectivity of CCR5-tropic HIVBaL and CXCR4-tropic HIVIIIB in vitro. We further demonstrate that this strain is genetically stable and can transiently colonize animal vaginal mucosa, while retaining important characteristics of the native bacterial phenotype.

The predominant mode of HIV transmission worldwide is via heterosexual contact, with the cervico-vaginal mucosa being the main portal of entry in women. The cervico-vaginal mucosa is naturally colonized with commensal bacteria, primarily lactobacilli. To address the urgent need for female-controlled approaches to block the heterosexual transmission of HIV, we have engineered natural human vaginal isolates of Lactobacillus jensenii to secrete two-domain CD4 (2D CD4) proteins. The secreted 2D CD4 recognized a conformation-dependent anti-CD4 antibody and bound HIV type 1 (HIV-1) gp120, suggesting that the expressed proteins adopted a native conformation. Single-cycle infection assays using HIV-1 HxB2 carrying a luciferase reporter gene demonstrated that Lactobacillus -derived 2D CD4 inhibited HIV-1 entry into target cells in a dose-dependent manner. Importantly, coincubation of the engineered bacteria with recombinant HIV-1 HxB2 reporter virus led to a significant decrease in virus infectivity of HeLa cells expressing CD4–CXCR4–CCR5. Engineered lactobacilli also caused a modest, but statistically significant, decrease in infectivity of a primary isolate, HIV-1 JR-FL . This represents an important first step toward the development of engineered commensal bacteria within the vaginal microflora to inhibit heterosexual transmission of HIV.

The present study aimed to characterize lactobacilli in vaginal fluid from 23 adult healthy women by using high-throughput DNA sequencing for identification of a large number of randomly selected colonies appearing on Rogosa and blood agar. The typing method was based on broad-range PCR of 16S rRNA gene variable regions V1 and V3, pyrosequencing, and classification of the fragments by alignment with NCBI-catalogued sequences and type strain sequences. Four major groups of sequences were found among the 402 isolates clearly corresponding to Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus iners and Lactobacillus jensenii when compared to the sequences obtained for type strains. Our results indicate that pyrosequencing of 16S rRNA gene fragments as used here is a fast and reliable method well suited for identification to the species level, even within the Lactobacillus acidophilus complex.

We examined the antimicrobial activity and composition of vaginal fluid.Vaginal fluid from preweighed tampons was assayed for pH, lactic acid, and antimicrobial polypeptides. The fluid was also fractionated by molecular filtration. Antimicrobial activity of whole fluid was determined against representative resident and exogenous microbes, and its fractions were tested against Escherichia coli.Vaginal fluids (5/5 donors) were permissive for Lactobacillus crispatus and vaginalis and Candida albicans, but not for Escherichia coli, Streptococcus group B, and Lactobacillus jensenii in three of five donors. The antimicrobial activity against E coli was predominantly in a3-kd fraction and correlated with both low pH and high lactic acid content. Compared with a matched pH buffer, lactic acid markedly suppressed the growth of E coli. Concentrated 2- or 5-fold, the protein-rich fraction was active against E coli.Vaginal fluid exerts selective antimicrobial activity against nonresident bacterial species. The activity is mediated by lactic acid, low pH, and antimicrobial polypeptides.

OA18.04 Background: Osel developed a recombinant live biotherapeutic product, MucoCept-CVN based on human vaginal Lactobacillus jensenii expressing the HIV-1 entry inhibitor Cyanovirin-N for prevention of vaginal transmission of HIV-1. We previously reported that macaques colonized with a live bacterial preparation in hydroxyethylcellulose showed a 63% reduction of SHIVSF162P3 acquisition followed repeated vaginal challenge. We formulated MucoCept-CVN as a fast dissolving vaginal tablet, which would allow for discreet, coital-independent, female controlled use. Tablets were potent (5.8 x 1011 colony forming units (CFUs)/gram) and stable for at least one year at 4°C and 25oC. Macaques were dosed with 1 tablet or 5 tablets delivered over 5 days. We examined colonization of the MucoCept-CVN bacteria in the rhesus macaque vagina at 14 and 21 days post dosing and found that 83% were highly colonized. Methods: Fast dissolving tablet were formulated (∼110 mg in a 1 x 1.5 cm mold). Tablet friability and dissolution were tested. Stability of the product was monitored for 1 year at 4°C and 25°C and for 2 months at 37°C. A single tablet or multiple tablets were administered to 18 rhesus macaques and vaginal colonization was monitored at 14 and 21 days post dosing by culture and PCR. Results: Tablets were potent and contained 5.8 x 1011 colony forming units (CFUs)/gram. The tablets were stable 1 yr at 4°C and 25oC and for 1 month at 37°C. The tablets maintained integrity when handled and dissolved quickly. High levels of colonization ∼108 CFU/vaginal swab of colonization were achieved in 15/18 macaques by day 21 post dosing. Two macaques had lower colonization levels ∼105 CFU/swab and in one macaque bacteria were detected by PCR only. Conclusions: MucoCept-CVN can be formulated and delivered as a conventional fast dissolving tablet dosage form. Colonization was achieved in 83% of macaques. The tablets were easy to handle and stable for>1 yr. This tablet formulation can be manufactured on large scale for ease of use.

Introduction The vaginal microbiome plays an important role in maintaining reproductive health throughout pregnancy. Despite the presence of an ‘abnormal’ vaginal microbial community being associated with an increased risk of preterm birth, interventional trials of antibiotics have failed to demonstrate significant benefit, which is likely due to a poor understanding of the vaginal microbiome in pregnancy. We sought to characterise the vaginal microbiome in uncomplicated pregnancies in a UK-based cohort. Methods Vaginal swabs were collected at time points across gestation (8–12, 20–22, 28–30 and 34–36 weeks) and the postpartum period (6 weeks) from 43 women delivered at term. Resident bacterial communities were determined using the Illumina MiSeq platform generated from bar-coded amplicons of 16S rRNA gene fragments. Results Individual vaginal microbiomes in pregnancy fall within four categories; i) Lactobacillus crispatus dominant, ii) Lactobacillus iners dominant, iii) Lactobacillus gasseri mixed with Lactobacillus jensenii and iv) Lactobacillus depleted with a diverse microbiome. Two pregnancy related trends were identified. In one group the Lactobacillus-dominated microbiome remains stable throughout pregnancy whereas in the other, an initially more diverse microbiome in the first trimester becomes Lactobacillus-dominated and less diverse in the second trimester but then reverts back to greater diversity in the third trimester and postpartum. Lactobacillus crispatus was associated with lower diversity and greater stability throughout pregnancy. Conclusion The vaginal microbiome during pregnancy tends to be Lactobacillus-dominated and less diverse in the period prior to fetal viability. Stability appears to be dependent upon the Lactobacillus species which may have implications for probiotic therapy.

Facultative anaerobic lactobacilli were recovered from the vaginas of 96.8% of 63 nonpregnant, healthy, premenopausal women. The predominant species were Lactobacillus jensenii, Lactobacillus acidophilus, and Lactobacillus casei. Of the women, 74.6% had hydrogen peroxide-producing lactobacilli, 22.2% had non-hydrogen peroxide-producing lactobacilli, and 3.2% had no lactobacilli. None of the 68 isolates had catalase activity. Some 68.2% of the isolates were inhibited by concentrations of less than or equal to 1% (wt/vol) of nonoxynol-9 (bactericidal for 73.3% of isolates, bacteriostatic for 26.7%). The remaining 31.8% could grow in all concentrations to 25% (wt/vol) of nonoxynol-9. All of the lactobacilli that were sensitive to nonoxynol-9 produced hydrogen peroxide whereas only 3 of 21 resistant strains were hydrogen peroxide producers. A significant correlation (P less than .001, chi 2 test) was found between hydrogen peroxide production and sensitivity to nonoxynol-9. It is suggested that the vaginal flora of spermicide users could be depleted of hydrogen peroxide-producing lactobacilli, possibly increasing susceptibility to urogenital infection.

Twelve strains isolated from the posterior fornix fluid of the human vagina were identified as Lactobacillus johnsonii, Lactobacillus acidophilus, Lactobacillus gallinarum and Lactobacillus crispatus based on numerical analyses of total soluble cell protein profiles and randomly amplified polymorphic DNA (RAPD)-PCR banding patterns. Five strains grouped with the type strains of Lactobacillus gasseri (DSM 20077T) and Lactobacillus jensenii (DSM 20557T) at r > or = 0.83 in one protein profile cluster, well separated from the other species included in this study. However, numerical analysis of the RAPD-PCR banding patterns of representative strains selected from the L. gasseri-L. jensenii protein cluster clearly indicated that they belong to two different species. Four strains (TV 1010, TG 1013, TV 1018T and TV 1045) grouped into another well separated protein profile cluster at r > or = 0.87. Strains selected from this cluster displayed very similar RAPD-PCR banding patterns and clustered at R2 > or = 0.78, separate from the other strains examined. Sequencing of the 16S rRNA of two representative strains, TV 1018T and TG 1013, of this group indicated that it represents a new member of rRNA group I Lactobacillus, which includes Lactobacillus delbrueckii, the type of the genus, and close relatives Lactobacillus acetotolerans, Lactobacillus kefiranofaciens, Lactobacillus iners, L. jensenii, L. crispatus, L. acidophilus, Lactobacillus helveticus, Lactobacillus amylovorus, Lactobacillus hamsteri, L. johnsonii, L. gasseri and Lactobacillus amylolyticus. The name Lactobacillus fornicalis sp. nov. is proposed for strains TV 1010 (DSM 13172), TG 1013, TV 1018T and TV 1045, with strain TV 1018T (= DSM 13171T = ATCC 700934T) as the type.

Lactobacillus acidophilus has been reported to be the predominant vaginal species. Vaginal lactobacilli isolated from 215 sexually active women were identified using whole-chromosomal DNA probes to 20 American Type Culture Collection Lactobacillus strains. Most women were colonized by L. crispatus (32%), followed by L. jensenii (23%), a previously undescribed species designated L. 1086V (15%), L. gasseri (5%), L. fermentum (0.3%), L. oris (0.3%), L. reuteri (0.3%), L. ruminis (0.3%), and L. vaginalis (0.3%). H2O2 was produced by 95% of L. crispatus and 94% of L. jensenii isolates, compared with only 9% of L. 1086V. Colonization by L. crispatus or L. jensenii was positively associated with being white (P.001), age/=20 years (P=.05), barrier contraceptive usage (P=.008), and lower frequency of bacterial vaginosis (P.001) and gonorrhea (P=.03). L. crispatus and L. jensenii, not L. acidophilus, are the most common species of vaginal lactobacilli.

Genetically modified commensal bacteria offer an unusual means for producing and delivering antiviral microbicides near or along specific organs where they are needed, according to Peter P. Lee at Stanford University in Stanford, Calif., and his collaborators. They engineered Lactobacillus jensenii, a commensal species that inhabits the vaginal tract of healthy women, to express cyanovirin-N (CVN), a protein that can inhibit HIV. The idea is to use this novel microbicide-producing commensal to protect women, who account for half of new cases, against becoming infected with HIV.

A correction is made to the article by Harris et al. [(2010) Acta Cryst. F66, 938–940]., The article by Harris et al. [(2010) Acta Cryst. F66, 938–940] is corrected.

It was hypothesized that Lactobacillus acidophilus might not be the dominant aerobic or microaerophilic Lactobacillus species in the human vagina, and that the dominant isolates are likely to express a variety of properties associated with colonization. To examine the question, vaginal swabs were collected from 100 healthy premenopausal women and cultured to detect the dominant aerobic or microaerophilic isolates of Lactobacillus. Only eight species were detected, with Lactobacillus jensenii being the dominant colonizers in the highest number (35) of women. Twelve different plasmid profiles were found amongst the isolates, but there was no association between plasmids and vaginal colonization or hydrogen peroxide production. Of the strains tested, few were resistant to the spermicidal agent nonoxynol-9. The data identify species within which strains may possess properties associated with maintenance of a healthy vaginal ecology.

High levels of Lactobacillus , the dominant genus of the healthy human vaginal microbiota, have been epidemiologically linked to a reduced risk of infection following exposure to the sexually transmitted pathogen Neisseria gonorrhoeae . In this work, a cell culture model of gonococcal infection was adapted to examine the effects of lactobacilli on gonococcal interactions with endometrial epithelial cells in vitro. Precolonization of epithelial cells with Lactobacillus jensenii , Lactobacillus gasseri ATCC 33323, or L. gasseri ATCC 9857 reduced gonococcal adherence by nearly 50%. Lactobacilli also inhibited gonococcal invasion of epithelial cells by more than 60%, which was independent of the effect on adherence. Furthermore, lactobacilli were able to displace adherent gonococci from epithelial cells, suggesting that these organisms have potential as a postexposure prophylactic. Thus, vaginal lactobacilli have the ability to inhibit gonococci at two key steps of an infection, which might have a significant effect in determining whether the gonococcus will be able to successfully establish an infection following exposure in vivo.

Urogenital infections are the most common ailments afflicting women. They are treated with dated antimicrobials whose efficacy is diminishing. The process of infection involves pathogen adhesion and displacement of indigenous Lactobacillus crispatus and Lactobacillus jensenii. An alternative therapeutic approach to antimicrobial therapy is to reestablish lactobacilli in this microbiome through probiotic administration. We hypothesized that lactobacilli displaying strong adhesion forces with pathogens would facilitate coaggregation between the two strains, ultimately explaining the elimination of pathogens seen in vivo. Using atomic force microscopy, we found that adhesion forces between lactobacilli and three virulent toxic shock syndrome toxin 1-producing Staphylococcus aureus strains, were significantly stronger (2.2-6.4 nN) than between staphylococcal pairs (2.2-3.4 nN), especially for the probiotic Lactobacillus reuteri RC-14 (4.0-6.4 nN) after 120 s of bond-strengthening. Moreover, stronger adhesion forces resulted in significantly larger coaggregates. Adhesion between the bacteria occurred instantly upon contact and matured within one to two minutes, demonstrating the potential for rapid anti-pathogen effects using a probiotic. Coaggregation is one of the recognized mechanisms through which lactobacilli can exert their probiotic effects to create a hostile micro-environment around a pathogen. With antimicrobial options fading, it therewith becomes increasingly important to identify lactobacilli that bind strongly with pathogens.

Background Lactobacillus species is an integral part of vaginal microbiota that maintains a healthy environment and plays an important role in preventing STI and HIV. We examined 20 women to investigate the difference in the diversity of Lactobacillus species present when the women are healthy or have Bacterial Vaginosis (BV). Method Between February and November 2010, samples from a total of 20 women attending Prerana Women9s Health Clinic were collected. Out of 20, 10 women were considered healthy and 10 women were diagnosed with BV based on Amsel9s Criteria. In addition, Gram stained smears of vaginal fluid were Nugent scored as negative, intermediate, or positive for BV. Based on the Nugent score criteria, nine were Positive, nine were negative and two showed “Intermediate” Nugent Score. Vaginal swabs were taken from the women with informed consent after ethical approval and grown in MRS broth. Gram positive Lactobacilli generating about 600–800 bp amplicon by16SrDNA PCR were further characterised by sequencing. Result Lactobacillus crispatus (40%) and Lactobacillus jensenii (40%) were the most common Lactobacillus species found in the vaginas of healthy women, the same Lactobacillus species found in healthy women in other countries. L crispatus was cultured from 40% of healthy women and none of women with BV. L jensenii , L gasseri , and L acidophilus were cultured from 40%, 10% and 10% of healthy women respectively; and none of the women with BV. Lactobacillus iners was not detected among healthy women or women with BV in our sample. Other organisms found among women were Staphylococcus epidermidis (60% among women with BV and 30% among healthy women), Streptococcus anginosus (40% among women with BV and 20% among healthy women). Some Corynebacterium spp were common among both women with BV and healthy women. Among the two women with “Intermediate” nugent score, one did not show growth of any Lactobacillus and in the other case there was growth of Lactobacillus salivarius . Conclusion Our findings showed Lactobacilli species present in healthy vagina of women in India do not differ from those reported from other countries. This information is useful for the development of microbicides for HIV prevention as well as better understanding of the reproductive health of women in India.

Several starch-hydrolyzing strains of Lactobacillus Beijerinck 1901 were isolated from cattle waste-corn fermentations. The isolates were facultatively anaerobic, gram-positive, nonmotile, nonsporeforming, rod-shaped organisms that grew singly and in short chains, produced lactic acid and small amounts of acetic acid but no gas from glucose, and did not exhibit oxidase-, catalase-, or nitrate-reducing activities. Growth occurred at 45°C but not at 15°C. dl-Lactic acid was synthesized from glucose. An extracellular amylolytic enzyme was formed. The organisms fermented amygdalin, cellobiose, esculin, fructose, galactose, glucose, maltose, mannose, salicin, starch, sucrose, and trehalose. Some strains fermented mannitol and lactose. Arabinose, gluconate, melezitose, melibiose, raffinose, rham-nose, ribose, and xylose were not fermented. The average guanine-plus-cytosine content of the deoxyribonucleic acid was 40.4 mol%. Reassociation values of 15% or less were obtained with the deoxyribonucleic acids of the starch hydrolyzers and those of the type strains of Lactobacillus acidophilus (Moro) Hansen and Mocquot 1970; Lactobacillus jensenii Gasser, Mandel, and Rogosa 1970; and Lactobacillus leichmanii (Henneberg) Bergey et al. 1923. The new species was differentiated from the three recognized species on the basis of starch fermentation, vitamin requirements, guanine-plus-cytosine contents, and stereoisomerism of lactic acid produced. It was concluded that these organisms represent strains of a hitherto unrecognized species for which the name Lactobacillus amylovorus is proposed. The type strain of L. amylovorus is NRRL B-4540.

Nucleic acid composition of 17 cultures representing the type or neotype strains of 15 named Lactobacillus species was studied. Nucleic acid characterization of these isolates was accompanied by a comparative study of conventional phenotypic reactions. The overall guanine plus cytosine mean deoxyribonucleic acid base composition ranged from 33 to 50% and genome sizes varied between 700 and 1500 X 10(6) daltons. Sporolactobacillus inulinus contained 2500 X 10(6) daltons of chromosomal deoxyribonucleic acid and was therefore similar in size to members of the genus Bacillus. Deoxyribonucleic acid hybridization at a temperature 15 degrees C below the thermal melting temperature confirmed the extreme molecular heterogeneity of many species. The genus can be divided into three major groups, each containing four or more species based on a combination of nucleic acid characteristics and conventional phenotypic reactions. Group I (33 to 39% guanine plus cytosine) contains Lactobacillus jensenii, Lactobacillus helveticus, Lactobacillus helveticus nov. ssp. jugurt, Lactobacillus acidophilus, Lactobacillus salivarius, and Lactobacillus sanfrancisco. Group II (42 to 48% guanine plus cytosine contains Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus casei, and Lactobacillus brevis, and Group III (48 to 50% guanine plus cytosine) contains Lactobacillus delbrueckii, Lactobacillus delbrueckii nov. ssp. bulgaricus, Lactobacillus delbrueckii nov. ssp. lactis, Lactobacillus delbrueckii nov. ssp. leichmanii, Lactobacillus fermentum, and Lactobacillus fermentum nov. ssp. cellobiosis.

The presence of Lactobacillus jensenii and Lactobacillus acidophilus has been studied in specimens from the rectum and vagina of the mother, from the mouth of the infant at the time of delivery, and from the mouth and rectum of infants six days of age. L. jensenii could be differentiated from other species of lactobacilli by the following combination of characteristics: production of only D-lactate, hydrolysis of arginine, and fermentation of cellobiose, galactose, and ribose, but not of lactose. L. jensenii and L. acidophilus were common inhabitants of the vagina. In spite of a contamination of the infant's mouth by L. jensenii and L. acidophilus during delivery, neither of these organisms became established in the mouth of the newborn infants.

The ability of Lactobacillus jensenii and Streptococcus 17SB to inbhibit Escherichia coli was demonstrated. The practical application of the inhibition was examined. Possible roles for several factors in the mechanism of inhibition by Lactobacillus jensenii and Streptococcus 17SB are briefly discussed. The effects of pH reached and acids produced by starters on the growth of Escherichia coli are described.

Labeled [3H]deoxyribonucleic acid (DNA) from Lactobacillus jensenii ATCC 25258 yielded a substantial amount of heteroduplexes only with DNA molecules from other strains of L. jensenii. A low yield of heteroduplexing was obtained between DNAs of L. jensenii and L. acidophilus, although both species had the same guanine plus cytosine content (36 mol%). The striking phenotypic similarity between L. jensenii and L. leichmannii was not reflected in their DNA base composition (36 and 50 mol% guanine plus cytosine, respectively), and, as expected, they produced a very low yield of hybridized DNAs. Although the phylogenic relationship between these different species of Lactobacillus cannot be deduced from their DNA relatedness, it can nevertheless be established from the structural similarity of their D-lactic dehydrogenases.