Use of Nucleic Acid Amplification Testing for Diagnosis of Extragenital Sexually Transmitted Infections.

Nucleic acid amplification testing (NAAT) is the preferred method to detect Chlamydia trachomatis and Neisseria gonorrhoeae, but no commercial tests are cleared by the U.S. Food and Drug Administration for use with extragenital swab samples. This study evaluated the performance of the Gen-Probe Aptima Combo2 assay (Aptima) and the Cepheid Xpert CT/NG assay (Xpert) to detect C. trachomatis and N. gonorrhoeae in rectal and pharyngeal samples from 224 men and 175 women reporting a history of anal receptive sexual intercourse. Discordant results between the NAATs were repeated using the assays APTIMA CT or APTIMA GC, which target alternate primers, as the confirmatory tests. C. trachomatis was detected from 59 rectal swabs and 8 pharyngeal samples, with 97.7% and 99.5% agreement between the two test systems, respectively. For C. trachomatis, Xpert was 95% sensitive (95% CI, 86 to 99%) and Aptima was 92% sensitive (95% CI, 81 to 97%) from rectal swabs, while both systems were 100% sensitive from pharyngeal samples. N. gonorrhoeae was detected from 30 rectal and 40 pharyngeal samples, with 99.5% and 97.5% agreement between the two test systems. The sensitivity of Xpert for N. gonorrhoeae from rectal swabs was 100% (95% CI, 88 to 100%) versus 93% (95% CI, 78 to 99%) for Aptima. From pharyngeal swab samples, Xpert was 98% sensitive (95% CI, 87 to 99.9%) versus 93% (95% CI, 80 to 98%) for Aptima. For C. trachomatis, neither system was >95% sensitive from the rectum, though both were >99.5% specific. For N. gonorrhoeae, Xpert had higher sensitivity than Aptima, but with more false positives from pharyngeal samples.

Susceptibility of bacterial vaginosis (BV)-associated bacteria to secnidazole compared to metronidazole, tinidazole and clindamycin.

Secnidazole, a 5-nitroimidazole with a longer half-life, is structurally related to metronidazole and tinidazole. For treatment of bacterial vaginosis (BV), secnidazole is a suitable single-dose oral drug having a longer serum half-life than metronidazole. The objective of this study was to evaluate the antimicrobial susceptibility of vaginal isolates of facultative and anaerobic bacteria to secnidazole, metronidazole, tinidazole and clindamycin. A total of 605 unique BV-related bacteria and 108 isolates of lactobacilli recovered from the human vagina of US women during the years 2009-2015 were tested for antimicrobial susceptibility by the agar dilution CLSI reference method to determine the minimal inhibitory concentration (MIC). The MIC90 (µg/mL) for secnidazole was similar to metronidazole and tinidazole for Anaerococcus tetradius (secnidazole: MIC90 2; metronidazole: MIC90 2; tinidazole: MIC90 4), Atopobium vaginae (32; >128; 128), Bacteroides species (2; 2; 2), Finegoldia magna (2; 2; 4), Gardnerella vaginalis (128; 64; 32), Mageeibacillus indolicus (2; 2; 2), Megasphaera-like bacteria (0.5; 0.25; 0.5), Mobiluncus curtisii (128; >128; >128) and Mobiluncus mulieris (>128; >128; >128), Peptoniphilus lacrimalis (4; 4; 4) and Peptoniphilus harei (2; 2; 4), Porphyromonas species (0.25; 0.5; 0.25), Prevotella bivia (8; 8; 8), Prevotella amnii (2; 1; 2) and Prevotella timonensis (2; 2; 2). In this evaluation, 14 (40%) of 35 P. bivia, 5 (14%) of 35 P. amnii and 21 (58%) of 36 P. timonensis isolates were resistant to clindamycin with MIC values of >128 µg/mL. Secnidazole, like metronidazole, was superior to clindamycin for Prevotella spp., Bacteroides spp., Peptoniphilus spp., Anaerococcus tetradius and Finegoldia magna. Clindamycin had greater activity against Atopobium vaginae, Gardnerella vaginalis and Mobiluncus spp. compared to the nitroimidazoles. All 27 Lactobacillus crispatus, 26 (96%) of 27 L. jensenii, 5 (19%) of 27 L. gasseri and 18 (67%) of 27 L. iners isolates were susceptible to clindamycin (MIC ≤2) while the MIC90 for all lactobacilli tested was >128 µg/mL for secnidazole, metronidazole and tinidazole. Secnidazole has similar in vitro activity against the range of microorganisms associated with BV compared to metronidazole or tinidazole. Further, secnidazole spares lactobacilli, a characteristic which is desirable in drugs used to treat bacterial vaginosis.

Survival of vaginal microorganisms in three commercially available transport systems.

Transport systems are used to collect and maintain the viability of microorganisms. Two Amies media based transport systems, BD CultureSwab™ MaxV(+) Amies Medium without Charcoal (MaxV(+)) and Fisherfinest® with Amies gel Transport Medium without charcoal (Fisherfinest®) were compared to a Cary-Blair media based transport system, Starswab® Anaerobic Transport System (Starswab®), for their capacity to maintain the viability of 17 clinical microorganisms commonly isolated from the vagina (Lactobacillus crispatus, L. jensenii, L. iners, group B streptococci, Candida albicans, Escherichia coli, Enterococcus faecalis, Atopobium vaginae, Peptoniphilus harei, Mycoplasma hominis, Gardnerella vaginalis, Dialister microaerophilus, Mobiluncus curtisii, Prevotella amnii, P. timonensis, P. bivia, and Porphyromonas uenonis). Single swabs containing mixtures of up to five different species were inoculated in triplicate and held at 4 °C and room temperature for 24, 48, 72, and 96 h (h). At each time point, swabs were eluted into a sterile salt solution, serially diluted, inoculated onto selected media, and incubated. Each colony type was quantified and identified. A change in sample stability was reported as a ≥1 log increase or decrease in microorganism density from baseline. Overall, the viability of fastidious anaerobes was maintained better at 4 °C than room temperature. At 4 °C all three transport systems maintained the viability and prevented replication of C. albicans, E. faecalis, GBS, and E. coli. Microorganisms having a ≥1 log decrease in less than 24 h at 4 °C included A. vaginae, G. vaginalis, and P. uenonis in Starswab®, L. iners, A. vaginae, and P. amnii in MaxV(+), and A. vaginae, G. vaginalis, P. bivia, and P. amnii in Fisherfinest®. At 48 h at 4 °C, a ≥1 log decrease in concentration density was observed for P. harei and P. amnii in Starswab®, G. vaginalis, P. bivia and P. uenonis in MaxV(+), and L. iners, P. harei, P. timonensis, and P. uenonis in Fisherfinest®. Overall, at 4 °C the viability and stability of vaginal microorganisms was maintained better in the Cary-Blair based transport system (Starswab®) than in the two Amies based transport systems.

Patterns of Extragenital Chlamydia and Gonorrhea in Women and Men Who Have Sex With Men Reporting a History of Receptive Anal Intercourse.

BACKGROUND: Screening for Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (GC) in men who have sex with men is risk based. Despite high frequencies of oral and receptive anal intercourse (RAI) among women, extragenital screening is not recommended. METHODS: Women (n = 175) and men who have sex with men (n = 224) primarily recruited from a sexually transmitted infection clinic reporting a lifetime history of RAI completed a structured questionnaire and clinician-collected swab samples from the rectum, pharynx, vagina (women), and urine (men). CT and GC were detected using 2 commercial nucleic acid amplification tests (Aptima Combo 2; Hologic, Inc, Bedford, MA; Xpert CT/NG, Cepheid Innovation, Sunnyvale, CA). RESULTS: The median age of the population was 26 years, 62% were white, and 88% were enrolled from a sexually transmitted disease clinic. Men were more likely than women to have GC (22.8% vs. 3.4%) and CT (21.9% vs. 12.6%). In men versus women, GC was detected in 16.5% versus 2.3% of pharyngeal swabs, 11.6% versus 2.3% of rectal swabs, and 5.4% versus 2.9% of urine samples or vaginal swabs. C. trachomatis was detected in 2.2% versus 1.7% of pharyngeal swabs, 17.4% versus 11.4% of rectal swabs, and 4.5% versus 10.3% for urogenital sites in men versus women. Overall 79.6% of CT and 76.5% of GC in men and 18.2% of CT and 16.7% of GC in women were detected only in the pharynx or rectum. CONCLUSION: Reliance on urogenital screening alone misses most of GC and CT in men and more than 15% of infections in women reporting RAI.

Mageeibacillus indolicus gen. nov., sp. nov.: a novel bacterium isolated from the female genital tract.

Three isolates of a bacterium recovered from human endometrium using conventional culture methods were characterized biochemically and subjected to 16S rRNA gene sequencing and phylogenetic analysis. Isolates were non-motile, obligately anaerobic, non-spore forming, asaccharolytic, non-cellulolytic, indole positive, Gram positive rods. Cell wall fatty acid profiling revealed C14:0, C16:0, C18:2 ω6, 9c, C18:1 ω9c and C18:0 to be the major fatty acid composition. The DNA mol % G+C was determined to be 44.2%. 16S rRNA gene sequence analysis revealed only 91% sequence similarity with the closest cultivated bacterial isolate, Saccharofermentans acetigenes. Based on genotypic and phenotypic data, all three isolates are considered to be members of the same species and data suggest it represents a novel genus and species in the order Clostridiales with an association with Clostridium rRNA cluster III within the family Ruminococcaceae. We propose the name, Mageeibacillus indolicus gen. nov., sp. nov. The type strain is BAA-2120(T) and CCUG 59143(T).

Bacterial vaginosis and the risk of trichomonas vaginalis acquisition among HIV-1-negative women.

BACKGROUND: The vaginal microbiota may play a role in mediating susceptibility to sexually transmitted infections, including Trichomonas vaginalis (TV). METHODS: Data were analyzed from HIV-1-seronegative women participating in HIV Prevention Trials Network Protocol 035. At quarterly visits for up to 30 months, participants completed structured interviews and specimens were collected for genital tract infection testing. T. vaginalis was detected by saline microscopy. Bacterial vaginosis (BV) was characterized by Gram stain using the Nugent score (BV = 7-10; intermediate = 4-6; normal = 0-3 [reference group]). Cox proportional hazards models stratified by study site were used to assess the association between Nugent score category at the prior quarterly visit and TV acquisition. RESULTS: In this secondary analysis, 2920 participants from Malawi, South Africa, United States, Zambia, and Zimbabwe contributed 16,259 follow-up visits. Bacterial vaginosis was detected at 5680 (35%) visits, and TV was detected at 400 (2.5%) visits. Adjusting for age, marital status, hormonal contraceptive use, unprotected sex in the last week and TV at baseline, intermediate Nugent score, and BV at the prior visit were associated with an increased risk of TV (intermediate score: adjusted hazard ratio [aHR], 1.73; 95% confidence interval [CI], 1.21-2.19; BV: aHR, 2.40; 95% CI, 1.92-3.00). Sensitivity analyses excluding 211 participants with TV at baseline were similar to those from the full study population (intermediate score: aHR, 1.54; 95% CI, 1.10-2.14; BV: aHR, 2.23; 95% CI, 1.75-2.84). CONCLUSIONS: Women with a Nugent score higher than 3 were at an increased risk for acquiring TV. If this relationship is causal, interventions that improve the vaginal microbiota could contribute to reductions in TV incidence.

Survival of Trichomonas vaginalis in wet preparation and on wet mount.

OBJECTIVES: Microscopy is an insensitive method for detection of Trichomonas vaginalis, but is widely used because it is both rapid and inexpensive. Diagnosis of trichomoniasis by microscopy requires that motile forms be identified in vaginal fluid samples. However, microscopy cannot always be performed immediately after sample collection. The objective of this study was to assess the impact of sample storage at room temperature on trichomonad motility. METHODS: Vaginal swab samples from 77 women positive for T vaginalis infection were collected to determine the impact of storage on wet preparations (swabs in plastic tubes with saline) and wet mounts (samples placed onto a glass slide with a coverslip). Samples were read at 400× every 30 min for the first hour and then once per hour thereafter until there were no motile trichomonads observed. RESULTS: For wet preparations, motility was 100% at 30 min, 99% at 60 min and decreased by 3%-15% each subsequent hour, with samples having a lower density of trichomonads losing motility more quickly. Trichomonad motility diminished more rapidly in wet mounts compared with wet preparations, with a 20% decrement in motility in 60 min. CONCLUSIONS: These data suggest that vaginal fluid samples for diagnosis of trichomoniasis should be stored in saline rather than on microscope slides until they are examined under the microscope and samples should be evaluated by microscopy within an hour of collection. These findings also suggest that clinical sites which cannot perform microscopy within 1 h of sample collection should consider the use of other diagnostic tests.

Comparative genomic analyses of 17 clinical isolates of Gardnerella vaginalis provide evidence of multiple genetically isolated clades consistent with subspeciation into genovars.

Gardnerella vaginalis is associated with a spectrum of clinical conditions, suggesting high degrees of genetic heterogeneity among stains. Seventeen G. vaginalis isolates were subjected to a battery of comparative genomic analyses to determine their level of relatedness. For each measure, the degree of difference among the G. vaginalis strains was the highest observed among 23 pathogenic bacterial species for which at least eight genomes are available. Genome sizes ranged from 1.491 to 1.716 Mb; GC contents ranged from 41.18% to 43.40%; and the core genome, consisting of only 746 genes, makes up only 51.6% of each strain's genome on average and accounts for only 27% of the species supragenome. Neighbor-grouping analyses, using both distributed gene possession data and core gene allelic data, each identified two major sets of strains, each of which is composed of two groups. Each of the four groups has its own characteristic genome size, GC ratio, and greatly expanded core gene content, making the genomic diversity of each group within the range for other bacterial species. To test whether these 4 groups corresponded to genetically isolated clades, we inferred the phylogeny of each distributed gene that was present in at least two strains and absent in at least two strains; this analysis identified frequent homologous recombination within groups but not between groups or sets. G. vaginalis appears to include four nonrecombining groups/clades of organisms with distinct gene pools and genomic properties, which may confer distinct ecological properties. Consequently, it may be appropriate to treat these four groups as separate species.

Incidence and epidemiology of Streptococcus pseudoporcinus in the genital tract.

Streptococcus pseudoporcinus, a beta-hemolytic microorganism first isolated from the female gastrourinary tract in 2006, cross-reacts with serogrouping kits for group B Streptococcus (GBS) and could be misidentified in the laboratory. The epidemiologic characteristics of this species have not been reported previously, but this organism is thought to be rare. Paired vaginal and rectal samples were collected from 663 nonpregnant women enrolled in a phase II clinical vaccine trial of a GBS type III capsular polysaccharide-protein conjugate vaccine, and isolates initially identified as S. pseudoporcinus were collected for further testing. A total of 120 isolates of S. pseudoporcinus were recovered from 36 unique individuals with 5.4% of 663 women having this organism recovered at least once during follow-up. All of these isolates cross-reacted with a commercially available GBS serogrouping kit. Women colonized with isolates confirmed as S. pseudoporcinus by genotypic and phenotypic methodologies were compared to women who were not colonized to determine whether there were any significant factors associated with acquisition of S. pseudoporcinus. Acquisition of S. pseudoporcinus vaginally and/or rectally was 36 per 846.0 women-years of follow-up for an annual incidence of 4 per 100 woman-years of follow-up. Acquisition of S. pseudoporcinus was independently associated with black women, being 30 to 40 years of age, recent Trichomonas vaginalis infection, primary or recurrent genital herpes, having bacterial vaginosis by Nugent criteria, and having had two or more male sexual partners since the last visit. This study suggests that S. pseudoporcinus is not rare, especially among black women, and could be misidentified as GBS.

Limitations of the dye-based method for determining vaginal applicator use in microbicide trials.

BACKGROUND: A dye-based method for determining applicator usage in microbicide trials has been developed to assess whether applicators have been exposed to vaginal fluid. Our objective was to evaluate this method on polypropylene HTI applicators that are being widely used in several effectiveness trials of microbicides. METHODS: Study participants enrolled in a clinical trial assessing SPL7013 (VivaGel) inserted gel intravaginally twice daily for 14 days and returned used and unused applicators. Before staining, smears were prepared from each participant-inserted applicator, Gram stained and assessed independently for the presence of vaginal cells and bacteria. Of the 169 participant-inserted applicators, 168 (99%) had vaginal cells identified by Gram stain. PARTICIPANT: Inserted applicators were stained with a 0.05% FD & C blue dye No. 1 solution and compared with 70 inserted positive control applicators and 70 unused negative control applicators. Intravaginally inserted applicators should stain turquoise, whereas unused applicators should not retain any stain. The individual responsible for labeling and preparing the applicators did not serve as an evaluator. RESULTS: : Under optimized conditions, the sensitivity and specificity ranged from 81% to 95% and 86% to 93%, respectively for single use and unused applicators. The dye-based method was only 47% to 77% sensitive for participant-inserted applicators obtained from women using gel twice daily. CONCLUSION: The dye test for HTI polypropylene applicators had a sensitivity of 47% to 95%, depending on the evaluator and whether gel was present in the vagina. The sensitivity was decreased with multiple gel applications. The dye-based method cannot be recommended for HTI polypropylene applicators to monitor product adherence.

Quantitative survival of aerobic and anaerobic microorganisms in Port-A-Cul and Copan transport systems.

Transport media should preserve the viability and stability of microorganisms in clinical specimens. In this study, the Port-A-Cul transport system and the Copan transport system without charcoal, both designed to preserve anaerobes, were evaluated. Dacron swabs were inoculated with two combinations of facultative and anaerobic organisms typically found in vaginal swab samples. Combination I contained Candida albicans, Escherichia coli, Enterococcus spp., group B streptococci, Lactobacillus crispatus, and Staphylococcus aureus. Combination II contained Lactobacillus iners, Peptoniphilus asaccharolyticus, Mycoplasma hominis, Prevotella bivia, Prevotella corporis, Porphyromonas asaccharolytica, Mobiluncus curtisii, Peptostreptococcus anaerobius, and Gardnerella vaginalis. Duplicate swabs were placed into the two transporters and held for 24, 48, 72, and 96 h at 4 and 24 degrees C. Both transporters maintained the viability of organisms better at 4 degrees C than at 24 degrees C. Prevotella bivia and Prevotella corporis had a loss of viability in both transporters at both temperatures. However, at 24 degrees C, there was a significantly greater loss of viability for Mycoplasma hominis, Prevotella bivia, Prevotella corporis, and Peptoniphilus asaccharolyticus when the organisms were stored in Copan transport medium than when they were stored in Port-A-Cul transport medium for 96 h (P < 0.002). Some organisms proliferated in the transport media, but when transporters were held at 24 degrees C for 96 h, a significantly greater increase in the concentrations of group B streptococci and Candida albicans, Escherichia coli, and Enterococcus spp. organisms in Copan medium than in Port-A-Cul medium was observed (P < 0.002). At room temperature, the Port-A-Cul system is superior to the Copan system with respect to the preservation of fastidious microorganisms and the prevention of the proliferation of facultative organisms.

Phase 1 Safety, Pharmacokinetics, and Pharmacodynamics of Dapivirine and Maraviroc Vaginal Rings: A Double-Blind Randomized Trial.

BACKGROUND: Variable adherence limits effectiveness of daily oral and intravaginal tenofovir-containing pre-exposure prophylaxis. Monthly vaginal antiretroviral rings are one approach to improve adherence and drug delivery. METHODS: MTN-013/IPM 026, a multisite, double-blind, randomized, placebo-controlled trial in 48 HIV-negative US women, evaluated vaginal rings containing dapivirine (DPV) (25 mg) and maraviroc (MVC) (100 mg), DPV only, MVC only, and placebo used continuously for 28 days. Safety was assessed by adverse events. Drug concentrations were quantified in plasma, cervicovaginal fluid (CVF), and cervical tissue. Cervical biopsy explants were challenged with HIV ex vivo to evaluate pharmacodynamics. RESULTS: There was no difference in related genitourinary adverse events between treatment arms compared with placebo. DPV and MVC concentrations rose higher initially before falling more rapidly with the combination ring compared with relatively stable concentrations with the single-drug rings. DPV concentrations in CVF were 1 and 5 log10 greater than cervical tissue and plasma for both rings. MVC was consistently detected only in CVF. DPV and MVC CVF and DPV tissue concentrations dropped rapidly after ring removal. Cervical tissue showed a significant inverse linear relationship between HIV replication and DPV levels. CONCLUSIONS: In this first study of a combination microbicide vaginal ring, all 4 rings were safe and well tolerated. Tissue DPV concentrations were 1000 times greater than plasma concentrations and single drug rings had more stable pharmacokinetics. DPV, but not MVC, demonstrated concentration-dependent inhibition of HIV-1 infection in cervical tissue. Because MVC concentrations were consistently detectable only in CVF and not in plasma, improved drug release of MVC rings is needed.

Transient detection of Chlamydial-specific Th1 memory cells in the peripheral circulation of women with history of Chlamydia trachomatis genital tract infection.

PROBLEM: Development of safe and effective Chlamydia trachomatis vaccines requires better understanding of the host immune responses elicited by natural infection. METHOD OF STUDY: Peripheral blood mononuclear cells isolated from women with or without history of genital tract chlamydial infection were stimulated with inactivated C. trachomatis elementary bodies (EB) in ELISPOT assays that enumerated frequencies of cells producing interferon (IFN)-γ or interleukin (IL)-17. RESULTS: IFN-γ-positive cells were highest among women sampled 30-60 days after diagnosis of C. trachomatis infection and treatment initiation, while the numbers of IFN-γ-positive cells were equally low among uninfected women and women sampled <30 or >60 days after diagnosis of infection. Conversely, IL-17-positive cell numbers were uniformly low among all participants. CONCLUSION: Dramatically reduced numbers of Chlamydia-specific Th1 memory cells in the peripheral circulation of study participants sampled more than 2 months after diagnosis, and initiation of treatment provides new insight into the results from C. trachomatis vaccine trials, in which immunization with EB provided only short-lived protection. Our results also suggest that an effective vaccine against this weakly antigenic intracellular pathogen will need to generate immunological memory more durable than that elicited by natural infection.

Development of a nonhuman primate model for Trichomonas vaginalis infection.

OBJECTIVE: Trichomoniasis, caused by Trichomonas vaginalis, is a prevalent sexually transmitted infection associated with increased risk of HIV infection. An animal model of T. vaginalis infection would enable scientists to further investigate trichomoniasis. STUDY DESIGN: Seven macaques (4 test vs. 3 control) were enrolled in a 2-week pilot study. Eight additional animals participated in a 2-arm (T. vaginalis vs. sham inoculated) crossover study lasting 5 weeks before treatment. In all, 12 Macaca nemestrina monkeys were challenged with a single intravaginal inoculation of 6.6 to 7.1 x 10(5) trichomonads (ATCC 50148). Vaginal culture (InPouch TV), colposcopy, microbiology, pH, and cervical cytokines were assessed at baseline, day 2, and weekly thereafter. RESULTS: Ten of 12 challenged animals tested positive for trichomoniasis for 2 weeks or longer. One animal tested positive on days 2 and 7 but negative thereafter. Only one animal was not infected. Oral metronidazole treatment (35 mg/kg per day for 3 days) resolved infection in all animals. Trichomoniasis infection did not lead to shifts in vaginal microbiology or pH. CONCLUSIONS: A single T. vaginalis inoculation results in persistent infection in the pigtailed macaque.

Colonization of the rectum by Lactobacillus species and decreased risk of bacterial vaginosis.

Lactobacilli colonizing the rectum may be a reservoir for vaginal lactobacilli. In a cross-sectional study of 531 females, vaginal and rectal colonization by lactobacilli were assessed by culture methods. A subset of isolates was identified to the species level by use of whole-chromosomal DNA probes. Lactobacillus crispatus (16%), L. jensenii (10%), and L. gasseri (10%) were the prevalent lactobacilli colonizing the rectums of 290 females. Only 13 (9%) of 147 females colonized by L. crispatus or L. jensenii vaginally and/or rectally had bacterial vaginosis (BV), compared with 12 (44%) of 27 females colonized by other H(2)O(2)-producing lactobacilli (P < .001). Cocolonization of the vagina and rectum by H(2)O(2)-producing lactobacilli was associated with the lowest prevalence of BV (5%), whereas females colonized only vaginally, only rectally, or at neither site had a successively increased risk of BV (P < .001). Lactobacillus species in the rectum may contribute to the maintenance of vaginal microflora.

Rectal applications of nonoxynol-9 cause tissue disruption in a monkey model.

BACKGROUND: Efforts to develop topical microbicide products have all but ignored evaluation for rectal use. GOAL: The goal of this study was to assess the effects of multiple rectal applications of Conceptrol (containing 4% nonoxynol-9) on flora and mucosal tissues in the pig-tailed macaque model. STUDY DESIGN: Monkeys (8 per group) received daily rectal applications of Conceptrol, placebo gel, or no product, for 3 days. At each visit, a preapplication rectal lavage specimen and swab specimen for microbiology and pH determination were collected. Conceptrol or placebo gel (2.5 ml) was then administered intrarectally. Fifteen minutes after application, samples were again collected. RESULTS: Gross observation of rectal lavage indicated sheets of epithelium 15 minutes after application of the nonoxynol-9 product. Histopathology of these samples revealed epithelial sheets with stroma attached. The presence of H(2)O(2)-producing lactobacilli remained relatively constant, whereas that of H(2)O(2)-producing viridans streptococci diminished in all nonoxynol-9-exposed animals in which these organisms were detected at baseline. CONCLUSIONS: Repeated applications of nonoxynol-9 disrupts the rectal mucosa of the pig-tailed macaque. The disruption of these tissues could have serious implications for an increase in likelihood of acquisition of sexually transmitted infection/HIV in humans.

Lactobacillus crispatus capsules: single-use safety study in the Macaca nemestrina model.

BACKGROUND: Lactobacillus crispatus is a part of the normal vaginal microflora of humans. GOAL: The goal of this study was to assess whether a capsule containing an H2O2-producing strain of L crispatus (CTV-05) would alter the vaginal microflora and/or epithelial tissues when applied intravaginally in the pig-tailed macaque model. STUDY DESIGN: Ten sexually mature female Macaca nemestrina were assessed at baseline for quantitative vaginal microbiology and vaginal pH and with colposcopy. One capsule containing 108 colony forming units of desiccated L crispatus CTV-05 was inserted into the vaginal fornix of each animal. Vaginal assessments were repeated on days 1 and 2 after capsule insertion. The L crispatus CTV-05 strain was identified with use of a DNA fingerprinting method. RESULTS: Before product use, four of 10 animals had detectable levels of H2O2-producing lactobacilli. L crispatus CTV-05 was detected in 1 of 10 animals on day 1 and in 3 of 10 animals on day 2 following insertion of the capsule. There were no tissue changes observed by colposcopy. Vaginal pH decreased in two animals colonized by CTV-05, from 7.0 at baseline to 4.5+/-0.5 on days 1 and 2 after product use. CONCLUSIONS: A single intravaginal application of capsules containing 108 L crispatus CTV-05 resulted in vaginal colonization in three of 10 animals 2 days after use. The absence of colposcopic changes in the vagina/cervical tissues indicates that L crispatus capsules are well tolerated.