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The Identification of Vaginal Lactobacillus Species and the Demographic
and Microbiologic Characteristics of Women Colonized by These Species
May A. D. Antonio,1 Stephen E. Hawes,3 1
Magee-Womens Research Institute and 2Department of Obstetrics,
and Sharon L. Hillier1,2 Gynecology, and Reproductive Sciences, University of Pittsburgh,
Pittsburgh, Pennsylvania; 3Human Papillomavirus Research Group,
University of Washington, Seattle

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

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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 va-
ginosis (P ! .001) and gonorrhea (P = .03 ). L. crispatus and L. jensenii, not L. acidophilus, are
the most common species of vaginal lactobacilli.

Establishing the identity of Lactobacillus species colonizing of L. acidophilus to identify human oral, intestinal, and vaginal
the vagina of women is of importance, because clinical studies Lactobacillus has been more a historic than a scientific desig-
have demonstrated an association between the presence of nation because of the poor reliability of existing tests used to
H2O2-producing strains of Lactobacillus and a decreased prev- differentiate Lactobacillus species [9]. Even 80 years ago, there
alence of gonorrhea, bacterial vaginosis (BV) [1], and human was uncertainty whether L. acidophilus characterized a group
immunodeficiency virus (HIV) infection [2–5]. However, in of related species or a single group of organisms that “under-
these studies lactobacilli have usually been identified only to goes transformation” [10].
the genus level because of the technical difficulties in the species Several investigators who were questioning the reliability and
identification of Lactobacillus. reproducibility of classic identification methods for Lactoba-
The identity of the predominant Lactobacillus species colo- cillus species sought more dependable identification protocols
nizing the vagina has been uncertain because of the unreliability [11, 12]. On the basis of DNA homology studies, the taxonomy
of classic identification methods, which employ sugar fermen- of lactobacilli has been under revision [12, 13]. Formerly, the
tation and other phenotypic assays. From these methods, var- species group of L. acidophilus comprised 6 DNA homology
ious lists of vaginal Lactobacillus species have been developed, groups that could not be distinguished biochemically [12]. Two
including any of the following: L. acidophilus, L. fermentum, of these homology groups are now L. crispatus and L. gasseri.
L. plantarum, L. brevis, L. jensenii, L. casei, L. cellobiosus, L. When DNA homology methods were used to evaluate the lac-
leichmanii, L. delbrueckii, and L. salivarius [6–8]. Of all these tobacilli from a group of 27 asymptomatic women, Giorgi et
species, L. acidophilus has been the vaginal lactobacillus most al. [14] identified L. gasseri, L. jensenii, and L. crispatus, not
widely accepted to be predominant . However, the general use L. acidophilus, as the predominant vaginal Lactobacillus species
colonizing asymptomatic women.
The present study was undertaken to identify which species
Received 10 August 1998; revised 3 August 1999; electronically published
12 November 1999.
of lactobacilli were present in a cross-sectional sample of 302
Presented in part: International Society for Sexually Transmitted Disease women, using DNA homology to American Type Culture Col-
Research Meeting, New Orleans, August 1995 (abstract 207). lection (ATCC) strains of lactobacilli. H2O2 production and the
Informed consent was obtained from each woman in a protocol approved
by the Human Subjects Committee at the University of Washington. In the
species specificity of this characteristic were determined. The
conduct of clinical research, human experimentation guidelines of the US distribution of Lactobacillus species colonizing women was also
Department of Health and Human Services were followed. assessed demographically and microbiologically.
Grant support: NIH (AI-31448, AI-38513).
Reprints or correspondence: Dr. Sharon L. Hillier, University of Pitts-
burgh, Dept. of Obstetrics, Gynecology, and Reproductive Sciences,
Magee-Womens Hospital, 300 Halket St., Pittsburgh, PA 15213-3180 Methods
In this study, 319 women visiting an adolescent medicine clinic
The Journal of Infectious Diseases 1999; 180:1950–6
q 1999 by the Infectious Diseases Society of America. All rights reserved. and 2 sexually transmitted disease clinics in Seattle were enrolled.
0022-1899/1999/18006-0025$02.00 A standardized questionnaire concerning demographic character-

[21]. Calcium alginate swabs or Dacron swabs were inserted into the For the DNA studies. were also inoculated and incubated at 367C U of Klenow enzyme. delbrueckii subspecies del- colony morphologies. vitamin The Random Primed DNA Labeling Kit (Boehringer Mann- K. hemin. casei subspecies casei 4646. trachomatis antigen and were examined for inclusions at 48–72 least 1 h. L. Gel electrophoresis and Vaginal swabs were removed from the transport medium and ethidium bromide staining were done to evidence genomic DNA used to inoculate Rogosa agar (Difco. acidophilus 521 and 4356. Gonococci were identified by sugar utilization. The slide and the transport milk until they were transferred to the Infectious Disease Labo- medium were delivered to the research laboratory within 12 h. trachomatis Luchansky et al. About 35 U of RNase A (Sigma. A probe was also made to a previously unidentified species dominant lactic acid peak as assessed by gas chromatographic anal. obic gram-negative rods were identified by use of Gram stain and confusus 10811.4 Slides were Gram stained and evaluated by use of the Nugent mL of 20% SDS was added and then incubated at 607C for 1 h. All lactobacilli were tested for the production of H2O2 in 1086V. The estimated specific activity of each probe was 2 3 10 9 a qualitative assay on a tetramethylbenzidine (TMB) agar plate disintegrations/min/mg. and they were further identified strains: L. Boston]) was incubated at 377C for at least 30 min. and ∼50 mCi [a32P] dATP [NEN Life Science in 5%–7% CO2 for a minimum of 72 h. Louis) was added h with an epifluorescence microscope [15]. by guest on September 5. A volume of 0. After 48 h of incubation in an anaerobic glove box at All DNA from patient isolates and ATCC Lactobacillus strains . L. to the solution. 30 mM Tris. L.oxfordjournals. Kellogg’s medium and either modified Thayer-Martin medium Sixteen units of a nonspecific protease type XIV from Streptomyces or enriched chocolate agar were streaked for the isolation of Neis. minutus 33267. An A7B agar plate and broths for dGTP. 1 mM EDTA. and the other agar to oxidize the TMB. L. and dTTP. L. [wt/vol] soluble starch. oris 49062. L. brueckii 9649. equal volume of chloroform. prereduced brucella agar with 5% sheep blood.2 M EDTA stopped the reaction. The specificity of this ratories. Detroit). C. designated ysis [20]. The pellet was resus- cloheximide-treated McCoy cells. gasseri 4963. A double volume of 95% ethanol was cies). ance readings at a wavelength of 260 nm. Anaer. Whole- Aerobic bacteria were first identified by use of Gram stain and chromosomal probes were made from 20 ATCC Lactobacillus colony morphologies and catalase. OR). L. leaving 302 evaluable subjects. jensenii 1951 istics and contraceptive history was administered. 1 mM Downloaded from http://jid. 25 ng denatured target genomic DNA. L. crispatus and L. and their in. [22]. 1 mg/mL of gentamicin. Troutdale. Indianapolis) was used to make whole-chromosomal probes blood bilayer Tween (HBT) agar plates (Prepared Media Labo. The DNA pellet was dissolved in flora. 5 mM EDTA.0) and disturbed flora. 1 h. prereduced laked-blood kanamycin agar. The following DNA isolation procedure was modified from 25 mg/mL of vancomycin. small gram. and 25 U/mL of nystatin. L. Prevotella. Two vaginal 367C–377C. pH 8. which was then incubated at 607C for 30 min. The H2O2 swabs were used to obtain samplings from the lateral vaginal wall. each Lactobacillus isolate was grown in cervix to obtain material for gonococcal and chlamydial cultures. griseus (Sigma) was added and was incubated at 377C for at least seria gonorrhoeae.0. according to the manufacturer’s protocol. The cells were washed in TES buffer (50 mM was cultured both in vials and in 96-well microtiter plates of cy. L. Lactobacillus isolates were stored at 2707C in litmus (MML Diagnostics. pigment production on HBT. L. solution sat in ambient temperature for 5 min. PYTSG broth (PY basal medium [20]. of Lactobacillus derived from a vaginal specimen. The [23] and Campylobacter [24] species. brevis 11577. L. L. and a score of 7–10 was considered consistent with stored at 47C. by their characteristic morphology on the A7B agar plate [18].JID 1999. and curved gram-variable rods (Mobiluncus spe. criteria [17]. 49540. 2 all prepared in-house. and production of pre. and 0. a score of 4–6 was considered consistent with intermediately a nominal amount of TE (10 mM Tris. hexanucleotide primers. and 2 human heim. 50 mM Tris. and examination of Gram-stained preparations [16]. The Columbia agar and 1 HBT plate were method has been demonstrated for identification of Mobiluncus incubated at 367C in 5%–7% CO2 for a minimum of 48 h. delbrueckii subspecies bulgaricus 11842.25 M EDTA. the agar plates were exposed to ambient air. vaginalis colony morphologies. ratory at the Magee-Womens Research Institute for DNA studies. buchneri by use of gas chromatography and biochemical tests [19]. Columbia 5% sheep isolation. For each whole-chromosomal remaining plates were incubated within an anaerobic glove box at probe. 2 mL of a reaction mixture containing random isolation of Ureaplasma urealyticum and Mycoplasma hominis [18]. 1% [wt/vol] dextrose. NaCI. A score of 0–3 was interpreted as consistent with normal added to precipitate the DNA. once with an equal volume of buffered phenol and once with an nerella species). above. causing the colonies of lactobacilli to swab for culture was placed into an Amies transport medium turn blue. The subcultured onto A7B agar.000 U of penicillin. salivarius subspecies salivarius 11741. 367C for a minimum of 5 days. The mycoplasmas were identified mentum 23271. rhamnosus 21052. Growth in broth was checked for Specimens for Chlamydia trachomatis cultures were transported purity by plating a drop onto a Columbia 5% sheep blood agar to the laboratory in 0. 1% Seventeen women were excluded because samples were not ob.0). 11579. The cell layers were stained with pended in lysis buffer (25% ultrapure sucrose. parabuchneri 49374. pH 8.02% [vol/vol] Tween 80) incubated tained for Gram staining or culture. L. blood agar. or Gard.180 (December) L. negative catalase test. and L. crispatus 33197. L. fer- ability to grow aerobically [19]. jensenii 25258. pH 8. pH 8. A score of 0–10 was assigned in light of the relative The solution was further incubated at 607C for 30 min. DNA concentration was determined using absorb- BV [17]. that was formed reacted with the horseradish peroxidase in the One swab was rolled onto a slide for a vaginal smear. the oxidase tests. L. The lysate was extracted negative or gram-variable rods (Bacteroides. St. OR). for 24–48 h in 6% CO2 at 377C. Tualitin.2 mL of sucrose-phosphate containing 2% plate (Prepared Media Laboratories) and incubating as described fetal calf serum. with the proportions of large gram-positive rods (lactobacilli). L. After the addition of 1 mL of 0. catenaformis 25536. The broths were further Products. addition of 10 U of proteinase K (Sigma). rum- Lactobacilli were identified to the genus level by Gram stain and inis 25644. a 20-mL reaction volume (consisting of 20 mM each dCTP. 2016 fluorescein-conjugated monoclonal antibodies to a species-specific EDTA.0) containing lysozyme and incubated at 377C for at C. addition of 2 mL of 0.

In brief. L. resulting in a different denominator. L. crispatus or L. Examples of 2 autoradiographs used in the DNA icantly decreased in women colonized by L. salivarius subspecies salicinius 11742. After being baked. At least 2 different Lactobacillus species simultaneously col- allowing 24 samples to be vertically arranged with 3 wells per onized 25 of the women. or douching did not appear to have washes were performed.1952 Antonio et al. pouch was incubated at 427C in a rocking incubator for at least 1 This comparison was chosen because nearly all of the L. each sample was aliquoted into 3 wells. N. Women with L. we marked the width of each well on the backside of the membrane. and 71% of them produced H2O2. OH) according to the manufac. johnsonii tobacillus strain we designated L. paracasei subspecies paracasei 27216. gonorrhoeae suggesting that visual interpretation of a positive hybridization was and C. Agreement between the 2 readers was 100%.05 turer’s instructions. buchneri. crispatus or L. Both L. cluded with each whole-chromosomal probe hybridization. 50%. Each 24-sample membrane was given a and L. or L. one was blinded to the identity of all unknown sam- likely to be white (72% vs. crispatus and L. jensenii were also less likely to have BV (9% vs. including 1 who was also colonized by L. About L. The specificity of the probe technique was excellent. delbrueckii.05) and more radiographs.008). table 1). jensenii were of the control positive band. species. gas- membrane from the slot blotter template piece. A dark band on the autoradiograph was any significant effect on the colonization of L. crispatus or L. a 1 : 10 volume of 3 N NaOH was added. acidophilus. jensenii with an isolate not homologous to any of the number and baked for 2 h at 807C. In some instances. respect to the presence of L. fermentum. crispatus and L. sample and control strips were laid flat. using a black seri. L. rhamnosus. L. probes are shown in figure 1. each probes used. P = . L. About one-third of the women were colonized by L. L. Keene. fermentum. was not revealed to this individual. The sample was incubated at 607C for 45 min. The microbiologic findings for the 302 women are shown in The location of a sample. If more than the intended species turned positive on fied by species of lactobacilli are shown in table 2.001) and users of barrier ples and control strains. The control filters were in. L. strains were identified. resulting in 6 strips. L. hybridization to L. not contrast. A Lac- 12315. an equal volume of 1 Results M ammonium acetate was added. and both L. read as positive if its intensity was similar to or greater than that jensenii. The following species 100 unknown isolates were tested against probes to all of the control were not recovered from the vagina of any of the women in Downloaded from http://jid. cris- h before the addition of whole-chromosomal probes. 2016 strains. overlapping one another. 27%. The heat-sealed nized by either species. women. 5%. L. Two individuals analyzed the auto. each with an identical sequence H2O2 (95% and 94%. After vacuuming was women. the slot blotter was disassembled. with the processing of DNA samples onto the nylon membranes. P < .180 (December) were slot-blotted onto nylon membranes of a Minifold II slot blot. About 1. The “blinded” individual did not work contraceptives (41% vs. Marital the control filter of ATCC Lactobacillus strains. ruminis. data were not available for some of overnight at 427C. A corner of patus and L. respectively. Before removing the 6 women. 1086V colonized 15% of the 33200. and one-fourth were colonized by L. less likely to be !20 years old (37% vs. jensenii col- sample. gasseri was recovered from only 5% of the women. trachomatis. L. gasseri whole-chromosomal jensenii (1% vs. only N. jensenii colonized a total of 154 women (51%). L. L. brevis 14869. jensenii. casei (table 1). L. and during onized 11 women. jensenii and L. parabuchneri. Either L. crispatus and L. P = . none of the unknown isolates having homology to 11 of the control catenaformis. jensenii strains produced H2O2. The control filters consisted of DNA from the ATCC strains listed above and from ATCC strains Lactobacilli were recovered from 215 (71%) of the 302 women L. buchneri 4005. L. women with L. acidophilus 4357. 1086V and L. L. gonorrhoeae infection was signif- not subjective. reuteri.53 standard saline the women. minutus 33267. crispatus or L. However. Of the sexually transmitted pathogens. permanent ink marker.5 mg 1086V. rhamnosus 21052.oxfordjournals. 1086V were coisolated from complete. plantarum 14917. and L. There were primed–labeled whole-chromosomal probe was added to the equil- ibrated membranes. table 3.15 M NaCl. L. L. salivarius. 61 lactobacilli-colonized women (20%) who did not have either After the addition of a probe. L. with this group: L. The slot blotter was connected to a by guest on September 5. jensenii and L. Pearson x2 tests were utilized to compare discrete variables with ter (Schleicher & Schuell. including women lacking lactobacilli. crispatus and L. 0. P < . 1086V were coisolated from 4 of sample DNA was aliquoted into each slot. 7. crispatus or L. L. higher-stringency status. jensenii with women not colo- was added to equilibrate the nylon membranes.1% SDS were Demographic characteristics and birth control usage strati- done at 657C. 69%. to a 400-mL total volume of TE (pH were considered statistically significant. Only 14 L. the membrane was incubated species.0) containing 5 mg of DNA. crispatus the pouch was cut open. casei. L. gasseri of samples and filter number. L. The wells of the slot blotter are arranged in a 3 3 24 fashion. fermentum 11739 and 14931. confusus. crispatus and L. brevis. L.03). ruminis 25644. JID 1999. L. oris. P values <. vaginalis. P = . and placed in an 8 3 12 inch heat-seal. The P values in tables 2 and 3 were obtained by comparing able pouch (Kapak. 49%. delbrueckii subspecies lactis 4797 and crispatus. dividing Nearly all of the L. In For each probe. unknown or control. gasseri.015 M sodium citrate) and 0. membrane was cut into 3 strips and then cut again in half. crispatus or L. citrate (0. and a 20-mL volume of random or L. 1086V isolates produced H2O2. Minneapolis). oral contraception. L. and only 1 woman each was colonized by L. only 9% of L. Both L. L. and L. jensenii. jensenii strains produced the wells in 2. Prehybridization solution [25] women with L. Three 45-min washes in 0. vacuuming. subspecies casei 393. gasseri 9857. Other isolated pairs included L. After the sample was cooled to ambient temperature.

ac- idophilus was not recovered from any of the women in this study. coli was less likely to be recov. jensenii (15% vs. who did not identify L. Most in species identification by use of phenotypic and DNA-based clinical isolates of L. we encourage the use of genomic-based methods for compared with only a few isolates of L.3) 0 (0) 0 (0) Lactobacillus 1086V 44 (15) 4 (9) 16 (36) No homology 13 (4) 4 (31) 6 (46) No lactobacilli 87 (29) NA 73 (84) Discussion NOTE. crispatus ATCC 33197 sample DNA (d1) on control filter. L. 77%. a L. L. L. b8. there was no significant decrease L. E.3) 0 (0) 0 (0) ered from women with L. P < . DNA homology group women colonized production BV present P < . and Mycoplasma hominis (22% vs. tobacillus species colonizing women of reproductive age. crispatus and L. gasseri 14 (5) 10 (7) 6 (43) plasma urealyticum. oris 1 (. Columns a–c include DNA samples of unknown vaginal lactobacilli. The identification of al. Identical sample portions of slot blot hybridization by guest on September 5. catenaformis 25536 (d4). anaerobic non. crispatus and L. asymptomatic women but did find strains homologous to L. jensenii 1953 . jensenii are the predominant vaginal Lac. plantarum 14917 (d3). gasseri ATCC 4963 whole-chromosomal probe hybridized only to DNA of ATCC L. NA. not applicable. Lactobacillus species detected among 302 women with or Gardnerella vaginalis (38% vs. [14].3) 0 (0) 0 (0) and Staphylococcus species. 1086V. In A. such as Urea. 27%. buchneri 4005 (d6). Specificity of each whole-chromosomal probe was evidenced when DNA samples of ATCC strains (identical to probe species) were positive. L.JID 1999.001) and to have decreased colonization of BV-related species: Table 1.001). acidophilus in a group of 27 benefits of these species in the vagina. reuteri 1 (. a7. with 154 Downloaded from http://jid. In light of the differences the species specificity of H2O2 production by lactobacilli. and L. L. This study confirms the DNA homology study of Giorgi L. [26] and Eschenbach et al. Group B Streptococcus and Enterococcus. L. crispatus or L. In B. b5. using whole-chromosomal probes made from American Type Culture Collection (ATCC) Lactobacillus crispatus strain 33197 (A) and ATCC L.3) 9 (0) 1 (100) L. vaginalis 49540 (d2). Both L. ruminis 1 (. L. and H2O2 production by the lactobacilli. L. Column d consists of DNA samples of ATCC Lactobacillus strains: L. L. Clinical vaginal lactobacilli homologous to probe were identified at a1–a4. 61%. crispatus and L. crispatus and L. 2016 women being colonized by 1 or both of these species. crispatus ATCC 33197 whole-chromosomal probe hybridized only to L. methods.oxfordjournals.01). (%) of the 302 women. . fermentum 1 (. [21] both reported that all of Figure 1. gasseri strain 4963 (B). and L. pigmented gram-negative rods (38% vs. a6. gasseri strains (d7. Values are no. Same samples were used and arranged in a similar fashion for each autoradiograph shown. d8) on control filter. 33%. gasseri 4963 (d7). without bacterial vaginosis (BV). jensenii allows us to focus on the potential et al. To our knowledge. jensenii produce H2O2. crispatus. this is the first published report to support gasseri. b4. jensenii a 69 (23) 65 (94) 5 (7) in colonization of other vaginal bacterial species. crispatus and L. crispatus 96 (32) 91 (95) 9 (9) Except for Escherichia coli. and c7. McGroarty et identifying lactobacilli to the species level. c6. One unknown vaginal Lactobacillus isolate was homologous to probe (c4). a L.001). jensenii were recovered from 11 women. jensenii. L. b7. P < . an- No. gasseri 9857 (d8). L. crispatus 33197 (d1). L. L. P < .180 (December) L. vaginalis 1 (.001). L.3) 0 (0) 0 (0) P = . of H2O2 aerobic black-pigmented gram-negative rods (11% vs. L.001). buchneri 11579 (d5). 84%.

As the primary species were also colonized by other species of lactobacilli. positive for lactobacilli tologic chorioamnionitis [30].12 women [28]. jensenii in 40% of the classic methods of identification between laboratories. Lactobacillus 1086V may be due to the unreliability and irreproducibility of the was coisolated with L. Chlamydia trachomatis 9 (6) 4 (7) 4 (5) . (%).87 This differs from the study by Nagy et al. jensenii colonized three-fourths of women L. 43% by onized by the same Lactobacillus species combination [32].oxfordjournals. jensenii or L. However. Nagy diagnosed with BV. producing lactobacilli. crispatus or L. crispatus and L. [27] demonstrated in vitro that L. Demographic data were not available for 1 woman.48 ence of a halide-peroxidase complex or alone. This activity was observed either in the pres- White race 111/154 (72) 31/60 (52) 42/87 (48) !. Concentrations of >107 cfu of vag- Downloaded from http://jid. [1] in a longitudinal study Ureaplasma urealyticum 91 (59) 32 (53) 59 (68) . crispatus or L. Demographic characteristics and birth control usage of vaginalis. a crispatus or L. compared with women not colonized by No.11 nonpregnant women lacking H202-producing vaginal lactoba. it was shown that Staphylococcus species 5 (3) 4 (7) 7 (8) . and 71% by Nagy et al. including women lacking lactobacilli.52 creased diagnosis of BV also has been reported in pregnant Barrier 57/138 (41) 16/54 (30) 19/78 (24) . crispatus and L. G. No. the percentage of L. crispatus or L. jensenii species isolated P cidal activity against G. stratified less frequently recovered from women colonized by these H202- by species of lactobacilli. a Includes women colonized by L. jensenii colonized 64% Neisseria gonorrhoeae 1 (1) 1 (2) 6 (7) . crispatus. b of lactobacilli that colonize the vagina and produce H2O2. jensenii with women not colonized BV by inhibiting the growth of BV-related microorganisms. In ad- women colonized by a species combination.001 bacilli was suggested by Hawes et al. 14 of whom onized by H202-producing lactobacilli. it is beneficial to know that none of the women colonized by both of these species were their identified L. jensenii strains. by these species. This prevalence is dition. crispatus Other None without BV. however. b Sixty-one women were colonized by species other than L. jensenii com- by these species. plement one another. [8]. [8]. vaginalis and Prevotella bivia (formerly Age !20 years 57/154 (37) 31/60 (52) 41/87 (47) .org/ by guest on September 5. Data are no. acidophilus or L. Microbiologic characteristics of women colonized or not colonized by lactobacilli. Compares women with L. anaerobic gram-negative rods and M. acidophilus isolates producing H2O2 unexpected. Contraception An association between H2O2-producing lactobacilli and de- Oral 29/138 (21) 16/54 (30) 8/78 (10) .05 Bacteroides bivius). Eschenbach et al. The vaginal colonization of L. Mycoplasma hominis 34 (22) 29 (48) 60 (69) !.01 cilli were twice as likely to develop BV than were women col. Anaerobic gram- tribution between asymptomatic women without BV and negative rods women with BV. a b or L.67 of nonpregnant women. 8% of the women were col- et al.001 Unmarried 129/154 (84) 48/60 (80) 81/87 (93) . . [26]. crispatus and/or L. which suggested Bacterial vaginosis 14 (9) 29/61 (48) 73 (84) !. Complete microbiologic data were not available for 1 woman. 14 of whom against these complications. JID 1999. reported H2O2 production by only 56% of onized by 11 Lactobacillus species. (%) of women with Lactobacillus isolated L.15 Enterococcus species 20 (13) 4 (7) 20 (23) . such as preterm birth [29] and his- NOTE. This is noteworthy.001 The clinical relevance of H202-producing isolates of lacto.008 Douche 12 times/ 5/146 (3) 5/58 (9) 6/86 (7) . Nonpigmented 59 (38) 38 (63) 75 (86) !. Although some Lactobacillus species may be jensenii combination accounted for nearly half of the Lacto- biochemically identifiable. inal lactobacilli per gram of vaginal fluid have been linked to crispatus.43 douching and having multiple sex partners. positive for characteristic/no.001 that there were no differences in the Lactobacillus species dis. 2016 were also colonized by other species of lactobacilli. crispatus and/or L. contradicting the claim that no 2 women are col- was reported to be 77% by McGroarty et al. Table 3. Eschenbach et al. crispatus or L. [8]. jensenii with women not colonized We do not know whether L. jensenii. jensenii or L.001 Black-pigmented 17 (11) 14 (23) 34 (39) !. (%) of women with Lactobacillus species these species. In addition. whereas only 12% of women with BV were col. Klebanoff et al. jensenii isolates were H2O2 producers.001 Gardnerella vaginalis 59 (38) 47 (78) 77 (89) !. since BV has been linked to month pregnancy complications. L.03 of 28 uninfected women and only 7% of 67 BV-positive women. Escherichia coli 23 (15) 13 (22) 26 (30) . The L.180 (December) Table 2. crispatus Other None were H2O2 produced by vaginal lactobacilli was the source of the a b c Characteristic or L. hominis were women colonized or not colonized by Lactobacillus species. Characteristic (n = 154) (n = 60) (n = 87) P c [21] reported that L. This disparity There were significantly fewer diagnoses of gonorrheal in- may result from the inability of biochemical assays to differ- entiate between the species formerly belonging to the L. L. aci- dophilus group. [21]. Similarly. jensenii was significantly linked to a lower prevalence of BV. including women lacking lactobacilli. There were 61 women colonized by species other than L. In this study. crispatus and L. an increased frequency of L. c a decreased incidence of preterm delivery [31]. jensenii during pregnancy may offer protection Includes women colonized by L. the discrepancy of the latter result bacillus combinations found. jensenii. jensenii may protect a woman from developing c Compares women with L.1954 Antonio et al. In this study. crispatus or L. jensenii species isolated onized by either species (table 3). After an adjustment was made for Group B Streptococcus 27 (18) 17 (28) 19 (22) .

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