Marilen Benner

MICROBIOTA AND ENDOMETRIAL HEALTH 147 6 Chen et al., 2017 Investigation of microbiota composition within female reproductive tract • 80 Surgery for conditions not known to involve infection ( hysteromyoma, adenomyosis, endometriosis, salpingemphraxis) Median 31 (range 22-48) Laparoscopy or laparotomy swabs, transcervical swabs 2 (1 obtained transcervically, 1 surgically) Dry swabs, pre- operative skin area, swabs of gloves used by surgeons. Swabs taken through cervical os to test influence of contamination compared to operative sampling, PBS/ physiological saline as diluent negative controls controls for sample processing, DNA extraction, and real-time qPCR ( the latter included additional ultrapure water control) Ion PGM™ system sequencing (Thermo Fisher) V4 - V5 Lactobacillus (30.6%), Pseudomonas (9.1%), Acinetobacter (9.1%), Vagococcus (7.3%) and Sphingobium (5%) Higher diversity in uterus compared to vaginal and cervical samples. High similarity between samples taken by entrance through cervical os versus surgical access through abdomen. Microbial profiles different than controls. High reproducibility. Determination of signature OTUs increasing in abundance from vagina to peritoneal fluid. High intra- individual correlation versus clear distinction between upper and lower RT also in inter- individual analysis Estimation of bacterial biomass of female reproductive tract Copy number obtained by qPCR species- specific for 4 major vaginal Lactobacillus species, divided by corresponding relative abundance based on 16S rRNA sequencing 1 Lowest biomass compared to other sites of reproductive tract (decreasing from 10^10 - 10^20 copies/sample at lower third of vagina towards posterior fornix and cervical canal and endometrium with 10^2 - 10^3 copies/sample). Much lower CT value of endometrial samples compared to negative controls. Higher diversity corresponds to lower biomass. Validation if sequencing live bacteria or debris • 15 Median 33 (range 24 - 41) Culturing of live bacteria (additional to 16S rRNA sequencing) 1 Positive cultures for 5 out of 15 samples; 8 different isolates belonging to 7 genera such as Lactobacillus, Staphylococcus and Actinomyces CT, cycle threshold; GnRH, gonadotrophic releasing hormone; GnRHa, gonadotrophic releasing hormone agonist; OTU, operational taxonomic unit; PGM, personal genome maschine; qPCR, quantitative PCR; RT, reproductive tract