This study aimed to characterise the microbial community within the endometrial cavity and endocervix in women with menorrhagia or dysmenorrhea. Paired endocervical and endometrial biopsy samples were collected from women undergoing operative hysteroscopy and/or laparoscopy. Samples were cohorted based on pathology, indications for surgery, and histological dating of the endometrium. Samples were interrogated for the presence of microbial DNA using a two-step next generation sequencing technology approach to exploit the V5-V8 regions of the 16S rRNA gene. Pyrosequencing revealed that the endocervix and endometrium share a minor microbial community, but that each site harbours a separate and distinct microbial population (p = 0.024). This was also the case for women with menorrhagia and dysmenorrhea (p = 0.017). Lactobacillus spp. were the most abundant microbial taxa present in 50% of the cohorts, and across all endocervical groups. Members of the genera Prevotella, Fusobacterium and Jonquetella were the most abundant taxa identified in samples collected from nulliparous women. It can be concluded that the female upper genital tract is not sterile. Microbial community profiling revealed differences in the endometrial microbial community profiles for: (1) the endocervix compared to the endometrium, and (2), women with menorrhagia versus dysmenorrhea. The distinct microbial community profiles in these women may offer insight into the pathology and clinical management of dysfunctional menstrual bleeding.

ObjectiveThere is a paucity of data characterizing the microbiota of the female upper genital tract, which controversially is described as a sterile site. We examine whether the fallopian tube harbours an endogenous microbial community.DesignThis prospective study collected from women undergoing total hysterectomy or salpingectomy-oophorectomy.SettingPrivate hospital gynaecology department.PatientsFallopian tubes were collected from women diagnosed with benign disease or for prophylaxis.InterventionsSamples were interrogated for the presence of microbial DNA using a next generation sequencing technology approach to exploit the V5 to V9 regions of the 16S rRNA gene.Main outcome measuresThe fallopian tube microbiota was characterized using traditional culture techniques and next generation sequencing.ResultsBacteria were isolated from 50% of cultured samples, and 100% of samples returned positive PCR results. Only 68% of the culture isolates could be confidently identified using automated diagnostic equipment in a clinical microbiology laboratory. Monomicrobial communities were identified only for cultured isolates (50%). Pyrosequencing revealed that all communities were polymicrobial. Lactobacillus spp. were not present in all groups, nor were they the most dominant isolates. Distinct differences in the microbial communities were evident for left compared to right fallopian tubes, ampulla versus isthmus, pre- and post- menopausal tissue, and in secretory phase fallopian tubes with and without Mirena intrauterine devices in situ (all p < 0.05).ConclusionThe female upper genital tract is not sterile. Distinct microbial community profiles in the fallopian tubes of healthy women suggest that this genital tract site supports an endogenous microbiota.

These data contribute further evidence that LNG-IUS is a safe and effective method for treating non-atypical EH. Whether LNG-IUS could provide a safe and cost-effective alternative to hysterectomy for atypical EH warrants further examination.

The endometrial cavity is an upper genital tract site previously thought as sterile, however, advances in culture-independent, next-generation sequencing technology have revealed that this low-biomass site harbors a rich microbial community which includes multiple Lactobacillus species. These bacteria are considered to be the most abundant non-pathogenic genital tract commensals. Next-generation sequencing of the female lower genital tract has revealed significant variation amongst microbial community composition with respect to Lactobacillus sp. in samples collected from healthy women and women with urogenital conditions. The aim of this study was to evaluate our ability to characterize members of the genital tract microbial community to species-level taxonomy using variable regions of the 16S rRNA gene. Samples were interrogated for the presence of microbial DNA using next-generation sequencing technology that targets the V5–V8 regions of the 16S rRNA gene and compared to speciation using qPCR. We also performed re-analysis of published data using alternate variable regions of the 16S rRNA gene. In this analysis, we explore next-generation sequencing of clinical genital tract isolates as a method for high throughput identification to species-level of key Lactobacillus sp. Data revealed that characterization of genital tract taxa is hindered by a lack of a consensus protocol and 16S rRNA gene region target allowing comparison between studies.

Aim: To determine whether cultivation-dependent and -independent analyses identifying fallopian tube bacteria were associated with visually observable microbial cells in situ using scanning electron microscopy. Patients: Fallopian tubes were collected from pre- and postmenopausal women undergoing salpingectomies for benign disease or as prophylaxis. Materials & methods: Fresh fallopian tube samples were processed for scanning electron microscopy to characterize fallopian tube ultrastructure. Histopathology was used to exclude fallopian tube abnormalities and for menstrual cycle staging of the endometrium. Results: Scanning electron microscopy revealed observable microbial cells in fallopian tube samples. Conclusion: In the absence of inflammatory pathology, the fallopian tube harbors a visually observable microbial population, which correlates with cultivation-dependent and -independent data, further refuting the sterility of this anatomical niche.

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We report a case of vaginal agenesis associated with a large fibroid arising from a Müllerian duct remnant. Vaginal agenesis is rare and infrequently encountered by gynaecologists. Its association with a pelvic mass is even more uncommon, and a medline search by the authors has failed to uncover another reported case. We provide the clinical details of this case and discuss the differential diagnoses.

Amplification of the V5 -V8 region of the 16S rRNA gene effectively speciates medically important 2 genital tract Lactobacillus species in the upper female genital tract 3 4 AUTHORS 5 ABSTRACT 31Background: The endometrial cavity is an upper genital tract site largely heralded as sterile, however, 32 advances in culture-independent, next generation sequencing technology have revealed that this site 33 harbours a rich microbial community which includes multiple Lactobacillus species. These bacteria are 34 considered to be the most common non-pathogenic genital tract commensals. Next-generation 35 sequencing of the female lower genital tract has revealed significant variation amongst microbial 36 community composition with respect to Lactobacillus sp. in samples collected from healthy and 37 diseased women. The aim of this study was to evaluate the ability of the 16S rRNA gene to 38 characterize genital tract lactobacilli to species-level taxonomy. 39Methods: Samples were interrogated for the presence of microbial DNA using two-step next 40 generation sequencing technology to exploit the V5-V8 regions of the 16S rRNA gene and compared 41 to standard speciation using qPCR. 42Results: The V5-V8 region of the 16S rRNA gene has sufficient sequence variation within frequently 43 encountered genital tract lactobacilli to allow accurate determination of relative abundance within 44 the community, and speciation for several key community members without completing additional 45 experimentation. 46 Conclusions: Next-generation sequencing of clinical genital tract isolates is an effective method for 47 high throughput identification to species-level of key Lactobacillus sp. 48 KEYWORDS 49 Lactobacillus sp.; genital tract; 16S rRNA; pyrosequencing; qPCR; speciation 50 51 IMPORTANCE 52