Poster Abstracts - S618



Articles

Vaginal microbiome and birth weight in a Zimbabwean cohort


Identification: Mugabe, Muchaneta

Credits: None available.

Vaginal microbiome and birth weight in a Zimbabwean cohort
 
Muchaneta Gudza-Mugabe1,2,6*, Enock Havyarimana1, Shameem Jaumdally1, Christina Balle1, Katie Lennard1, Andrew Tarupiwa2, Fortunate Mugabe4, Rooyen Mavenyengwa3 Lindi Masson1, Heather Jaspan1,5
1University of Cape Town, South Africa; 2National Microbiology reference laboratory, Zimbabwe; 3University of Zimbabwe; 4Harare Central Hospital, Zimbabwe; 5Seattle Children's Research Institute, USA; 6Letten Foundation Research Centre
 
Introduction
Poor birth outcomes are a major cause of child morbidity and mortality, particularly in developing countries.  Birth weight is an objective measure of birth outcomes, encompassing both preterm and growth restricted pregnancies. This study investigated the relationship between the vaginal microbiome during pregnancy and low birth weight (LBW).
 
Methodology
Vaginal swabs were collected from 420 pregnant Zimbabwean women between 13-35 weeks of gestation who were then followed until delivery.  Nugent score was used to determine bacterial vaginosis (BV) status. DNA was extracted and sequencing of the 16S rRNA gene V4 hypervariable region was performed using Illumina MiSeq. Cytokine analyses were performed using Luminex.
 
Results
There were 233 singleton pregnancies with complete sequencing and birth outcome data, of which 41 (17.6%) resulted in the delivery of LBW infants (<2500g). Women who delivered LBW infants had significantly lower vaginal alpha diversity than women who experienced normal births and were less likely to have BV compared to term births (Median Shannon indices 0.67 vs 1.05; p=0.02). However, no significant differences in beta diversity of Bray Curtis distances were evident between women who delivered LBW versus normal infants (ADONIS p= 0.06). The vaginal microbiota of the cohort clustered into three community types using Fuzzy clustering (C1 and C2 were Lactobacillus iners and crispatus dominant, respectively, whereas C3 was Gardnerella vaginalis dominated). Women who delivered LBW infants clustered mainly with C1.  Vaginal concentrations of multiple inflammatory cytokines and growth factors were significantly lower in women with LBW infants. In a multivariate model, preterm delivery, community type, pregnancy-induced hypertension and growth factors were independently associated with risk for LBW, after adjusting for important confounders such as maternal age.
 
Conclusion
In this cohort of Zimbabwean women, multiple factors measured during pregnancy were found to be predictive of birth weight, including preterm delivery, pregnancy-induced hypertension, a vaginal microbiome dominated by L. iners and a deficit of cellular growth factors.


Overview of Vaginal Practices in Nigerian Women, effects on Microbiome and Cervical Cancer


Identification: Nwaokorie, Francisca

Credits: None available.

Overview of Vaginal Practices in Nigerian Women, effects on Microbiome and Cervical Cancer
 
Francisca Nwaokorie*1, Oliver Ezechi2, Folasade Ogunsola3
1Department of Medical Laboratory Science; Faculty of Basic Medical Sciences, College of Medicine, University of Lagos, Nigeria; 2Clinical Science Division, Nigerian Institute of Medical Research, Yaba, Lagos, Nigeria, 3Department of Medical Microbiology and Parasitology, Faculty of Basic Medical Sciences, College of Medicine, University of Lagos, Nigeria.
*Corresponding author
 
There is increasing evidence that vaginal microbiome can influence women's reproductive health.  The presence of certain species are associated with the maintenance of a healthy vagina.  Disruption of the microbiota, due to unprotected sexual activities, as well as intravaginal practices may put women at a higher risk of acquiring sexually transmitted infection including Human immunodeficiency Virus/Acquired immune Deficiency Syndrome (HIV/AIDS) and cervical cancer. Prevalent high-risk HPV infection have been identified in Nigerian women with or without HIV infection. However, the pattern of microbiome and its association with cervical cancers and HIV infection which is endemic in Nigeria is not fully established. Vaginal products and or vaginal practices are used in cleansing, tightening and or drying the vagina. To what extent this contributes to vaginal microbiome is not well understood. This review provide information on composition of cultivable and non culturable bacterial species found in Nigerian women in health and disease conditions. It also assessed factors including traditional practices that may lead to changes in microbiome of women of reproductive age. Such requires studies to see what maybe their roles in vaginal microbiome and possible relationship to high risk human papilloma virus.  
 


Non-human primate model for genital tract microbiome research


Identification: Obiero, Jael

Credits: None available.

Non-human primate model for genital tract microbiome research
 
Jael Obiero1,2, Peter G. Mwethera1
1Institute of Primate Research, Department of Reproductive Health & Biology; 2Jomo Kenyatta University of Agriculture & Technology, Department of Medical Microbiology
 
Background: The vaginal microbiome is believed to influence host health by providing protection from pathogens and influencing reproductive outcomes such as fertility and gestational length. Knowledge of the composition of vaginal microbial ecosystem is essential for understanding the etiology, prevention, and treatment of vaginal diseases. A baboon model has been used to provide detailed understanding of reproductive physiology and immunology applicable to women. However, little is known about the composition of its vaginal microbial ecosystem.
 
Methods: Gram-stain and Nugent scores were used for assessment of baboon vaginal microbial flora. Biochemical identification and analysis of isolates were performed using the Analytical Profile Index kits and identification software.
 
Results: Species of Lactobacilli, Staphylococci, Clostridia, Bacilli, Corynebacteria, Gram-negative rods, other Gram-positive rods, cocci and Candida, were isolated. Healthy vaginal microbiota consisted mainly of lactobacillus morphotypes. Animals with high Nugent scores had increased number of Gram-positive cocci and variable rods, with increased number of Gram-negative morphotypes.
 
Conclusion: The baboon vaginal microbiota is heterogeneous in terms of species composition and is typified by a scarcity of lactobacilli. Characterisation of baboon vaginal microbial communities, their interactions and impact on reproductive outcomes warrant investigation.


Prevalence, antimicrobial susceptibility, serotypes and risk factors of group B streptococcus rectovaginal isolates among pregnant women at Kenyatta National Hospital


Identification: Salano, Jisuvei

Credits: None available.

 

Prevalence, antimicrobial susceptibility, serotypes and risk factors of group B streptococcus rectovaginal isolates among pregnant women at Kenyatta National Hospital
 
Jisuvei Clayton Salano1,2, Maina Anne Njeri1, Osoti Alfred1  
1University of Nairobi, 2Kenyatta National Hospital
 
Estimates of group B streptococcus (GBS) disease burden, antimicrobial susceptibility and its serotypes in circulation among pregnant women in many developing countries including Kenya, are limited, yet these data are required for prophylaxis and treatment of infections due to GBS. We evaluated the rectovaginal prevalence, antimicrobial susceptibility, serotypes and factors associated with GBS colonization among pregnant women receiving antenatal care at Kenyatta National Hospital (KNH) between August and November 2017. In this cross sectional study, 292 consenting pregnant women between 12 and 40 weeks of gestation were enrolled. Interview-administered questionnaires were used to assess risk factors associated with GBS colonization. Two swabs; one from the anorectal canal the other from the lower vagina were collected and cultured on Granada agar for GBS isolation. Positive colonies were tested for antimicrobial susceptibility to penicillin G, ampicillin, vancomycin and clindamycin. Serotyping was done using Immulex Strep-B kit. We used logistic regression to identify factors associated with GBS colonization. Data analysis was done using STATA® version 13. P values of <0.05 were considered significant. The median age of study participants was 30 years (IQR 26-35) with a median gestational age of 35 weeks (IQR 30-37). The prevalence of GBS in this study was 20.5%. Isolates were most resistant to penicillin G (72.4%) followed by ampicillin (55.2%), clindamycin (30.4%) and vancomycin (24.1%). All ten GBS serotypes were isolated. Serotype Ia was the most prevalent (75.9%) while serotype VIII (44.2%) was the least occuring. 37 (69.8%) participants carried more than one GBS serotype. None of the risk factors were associated with GBS colonization. The prevalence of GBS is high among mothers attending antenatal clinic at KNH. There is high prevalence of GBS isolates resistant to commonly prescribed intrapartum antibiotics hence other measures like GBS vaccination is a potentially useful approaches to GBS prevention and control in this population. Screening of pregnant mothers for GBS colonization should be introduced.
 
Funding source: National Research Fund-Kenya
 

 


Hormonal Contraception is Associated with Lactobacillus iners Dominated Cervicovaginal Microbiota in Reproductive-Age Black South African Women


Identification: Onywera, Harris

Credits: None available.

Hormonal Contraception is Associated with Lactobacillus iners-Dominated Cervicovaginal Microbiota in Reproductive-Age Black South African Women
 
Harris Onywera1,2, Anna-Lise Williamson1,2,3, Zizipho Z. A. Mbulawa1,2,3,4, David Coetzee5, Tracy L. Meiring1,2*
1Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Cape Town, South Africa; 2Division of Medical Virology, Department of Pathology, Faculty of Health Sciences, University of Cape Town, South Africa; 3UCT-MRC Clinical Gynaecological Cancer Research Centre, University of Cape Town, Cape Town, South Africa; 4Center for HIV & STIs, National Institute for Communicable Diseases, National Health Laboratory Service, Johannesburg, South Africa; 5Center for Infectious Disease Epidemiology and Research, School of Public Health and Family Medicine, University of Cape Town, South Africa
*Corresponding author
 
Cervicovaginal microbiotas (CVMs) have a profound influence on the women's reproductive health. A CVM dominated by Lactobacillus spp. is thought to be a biomarker for cervicovaginal health. Complex and undesirable imbalances in the CVMs predisposes women to bacterial vaginosis (BV), the most common vaginal syndrome among reproductive-age women. BV has been associated with infertility and increased susceptibility to infections such as genital human papillomavirus (HPV), which is causally associated with cervical cancer. Despite the high health burden of HPV in Africa and evidence that a majority of women of African ancestry lack Lactobacillus-dominated CVMs, the CVMs of African women remain understudied. Here, the CVMs of Black South African women with and without HPV were characterized and associated with the participants' metadata. The CVMs of 62 reproductive-age women were profiled from cervical DNA by using Ion Torrent sequences from the V4 hypervariable region 16S rRNA gene. The CVMs were analyzed using QIIME, UPARSE, and metagenomeSeq tools. Associations of CVMs with participants' categorical and continuous variables were computed by Chi-square/Fisher's exact and Kruskal-Wallis tests, respectively. Twenty three women (37.1%) were HPV-positive. Twenty five women (40.3%) were on hormonal contraception. The CVMs clustered into three discrete community state types (CSTs): CST I (n=24, 38.7%) and CST II (n=3, 4.8%) that were dominated by Lactobacillus iners and an unclassified Lactobacillus species, correspondingly; and CST III (n=35, 56.5%) that was enriched with an array of heterogeneous BV-associated bacterial taxa, predominantly Gardnerella, Prevotella, Sneathia, and Shuttleworthia. CST III was associated with BV (p=0.001). Neither CST nor bacterial diversity was associated with HPV infection. Women in CST I were more likely to be on hormonal contraception, especially progestin-based, compared to women in CST III (odds ratio: 5.2 [95% CI 1.6-17.2]; p=0.006). The majority of the women had CVMs not dominated by Lactobacillus. Additional studies are needed to examine whether these CVMs represent abnormal, intermediate or variant states of health. Our findings on the association of hormonal contraception with L. iners-dominated CVMs warrants further investigation and may have implications on personalized microbiota-based diagnostics and probiotics that promote reproductive health.
 


Wishing for Wellness’s “Taxi Talks” – Using a forum theatre approach to talk to adolescents about reproductive health


Identification: Passmore, Jo-Ann

Credits: None available.

Wishing for Wellness's “Taxi Talks” - Using a forum theatre approach to talk to adolescents about reproductive health
 
Ntomboxolo Makhutshi1, Bobbie Fitchen2, Joanna Glanville2, Zandile Ciko3, Margaret Jacks4, Felicity Hartley2,3, Dante Robbertze5, Katherine Gill5, Glenda Gray6,7, Smritee Dabee3, Shameem Z. Jaumdally,3 Shaun Barnabas3,5, Linda-Gail Bekker3,5, Linzi Rabinowitz8, and
Jo-Ann S. Passmore3,9,10
1Center for Theatre, Dance and Performance Studies, University of Cape Town, Cape Town, South Africa; 2Dynamic Facilitators, Cape Town, South Africa; 3Institute of Infectious Diseases and Molecular Medicine, University of Cape Town, Cape Town, South Africa; 4Independent evaluator; 5Desmond Tutu HIV Foundation, University of Cape Town, Cape Town, South Africa; 6Peri-natal HIV Research Unit (PHRU), Soweto, South Africa; 7South African Medical Research Council (SAMRC), Cape Town, South Africa; 8Empathy Trust, Cape Town, South Africa; 9SAMRC-UCT Gynaecological Cancer Research Centre, University of Cape Town, Cape Town, South Africa; 10National Health Laboratory Service, Cape Town, South Africa
 
WISH'ing-for-Wellness (W4W) employed participatory drama-based methods to create a safe space for adolescent and young women to understand their sexual and reproductive health and rights. The project emerged from research which found high rates of sexually transmitted infections but poor health seeking behaviour in young women from socio-economically deprived townships in Cape Town, South Africa. To address the silence, misinformation and stigma around reproductive health, we constructed a life-sized taxi to playfully reflect the public realm in which adolescent and young women in South Africa live, using an adaption of Boal's forum theatre (theatre of the oppressed) to create the platform - called “Taxi Talks” - to confront social and gender discourses, including sex, 'blessers' (sugar daddies), family life, desire, and love. Most significant change (MSC), after action reviews (AAR), and journals kept by the facilitators evaluated the impact of this approach. “Taxi Talks” demonstrated that the young women's courage to challenge the conservatism they face in their communities around sexual reproductive health. Some of the stereotypical characters the young women brought to life in Taxi Talks included “the judgmental and bossy clinic sister who had a baby when she was a teenager”, and “the pregnant young woman that will give the baby to her mother to look after”. These characters allowed some participants to see that they did not want to be like that. With this burden of STIs/BV, poor school-based programmes, and risky behaviour early during their reproductive lives, developing adolescent-focused engagements, such as Taxi Talks and forum theatre, centred on reproductive and sexual health is critical.


Sexual behaviours impact the vaginal microbiota of women who have sex with women


Identification: Plummer, Erica

Credits: None available.

Sexual behaviours impact the vaginal microbiota of women who have sex with women
 
Plummer EL1, Vodstrcil LA1,2, Murray GL3, Tabrizi SN3, Garland SM3, Fairley CK1, Tan A3, Law M4, Hocking JS2, Bulach DM5, Philip G5, Bradshaw CS1,2
1Central Clinical School, Monash University; Melbourne Sexual Health Centre, Alfred Health; 2Melbourne School of Population & Global Health, The University of Melbourne; 3The Royal Women's Hospital; 4Kirby Institute, UNSW Australia; 5Melbourne Bioinformatics, The University of Melbourne
 
We investigated the impact of sexual behaviors on the vaginal microbiota (VM) of women-who-have-sex-with-women (WSW) participating in a 2 year cohort study. Women self-collected high vaginal swabs and completed a behavioral questionnaire every 3 months for 24 months or until incident bacterial vaginosis (BV). We characterized the VM using 16S-rRNA gene sequencing of the V3V4 region. Community state types (CSTs) were identified using hierarchical clustering. Bacterial diversity was calculated using the Shannon diversity index and instability of the VM was assessed using change of CST and Bray-Curtis dissimilarity between consecutive longitudinal specimens. The impact of behaviors on diversity and instability of the VM was determined using multivariate regression models. Linear discriminant analysis effect size was used to identify bacteria associated with exposure to a new sexual partner. 360 specimens from 100 women were included in analyses. The VM clustered into 5 CSTs: 3 dominated by Lactobacillus spp. (CST1 L. crispatus; CST2 L. crispatus and L. iners; CST3 L. iners), one abundant in Gardnerella vaginalis and one of mixed bacteria. Exposure to a new sexual partner increased bacterial diversity (Adj coef=0.33,95%CI:0.11,0.54) and instability of the VM, both in terms of change of CST (AOR=2.69,95%CI:1.37,5.28) and increased Bray-Curtis dissimilarity (Adj coef=0.22,95%CI:0.12,0.32). Sex with a new partner increased the abundance of bacteria often seen in BV including G. vaginalis, Megasphaera and BVAB1 (p<0.05). Conversely, no sex/sex in established ongoing relationships was associated with a favorable vaginal microbiota abundant in L. crispatus. Sex with a new partner markedly reshapes the VM of WSW by increasing the diversity and abundance of potentially pathogenic bacteria.
 
Funding: National Health and Medical Research Council (NHMRC) Program Grant #1071269 and NHMRC Project Grant #1020457
 


Vaginal Microbiome of premenopausal Indian women: A comparison of healthy and dysbiotic flora


Identification: Pramanick, Rinku

Credits: None available.

Vaginal Microbiome of premenopausal Indian women: A comparison of healthy and dysbiotic flora
 
Rinku Pramanick1, Niranjan Mayadeo2, Himangi Warke2 and Clara Aranha1   
 1 ICMR-National Institute for Research in Reproductive Health, Parel, Mumbai
2 Department of Obstetrics and Gynecology, Seth G.S. Medical College & KEM Hospital, Parel, Mumbai
 Presenting author email: rinku.pramanick@gmail.com
Corresponding author email: aranhac@nirrh.res.in
 
The vaginal microbiome plays a critical role in determining the progression of female genital tract infections. Since the vaginal microbiome varies with geography and ethnicities, deciphering the lacking information on Indian women is needed. We aimed to decode the vaginal microbial architecture of women with healthy and dysbiotic flora. To achieve this, we sequenced 16S rRNA V3-V4 region (HiSeq Illumina) of DNA extracted from vaginal swabs collected from women with bacterial vaginosis (BV) (n=10) and healthy controls (n=10) of 18-45yrs. The abundance profile of 20 samples had 876 OTUs from 33 families and 45 genera. Rarefaction analysis showed higher number of species in normal flora compared to BV. Alpha diversity as measured by Shannon diversity revealed normal flora had less diverse communities compared to BV. Beta diversity comparison using Bray Curtis indicated distinct microbial communities between normal and BV flora. The most abundant Phylum that showed significant difference between normal and BV samples were Firmicutes (p=0.0004) and Actinobacteria (p=0.009) where Firmicutes were abundant in normal flora. A significant difference was observed for relative proportions of Lactobacillus, Gardnerella, Aerococcus, Brevibacterium between the groups. Lactobacillus abundance decreased significantly in BV flora (14.57%), as compared to normal flora (83.25%) whereas the abundance of Bifidobacterium was observed increased from 12.0% in normal flora to 45.25% in BV. Besides, increased levels of Klebsiella, Sneathia, Coriobacteriaceae and Prevotella was noted in BV. L.crispatus was exclusively present in normal flora whereas L.iners was detected from both the groups with 80% and 100% prevalence in normal and BV flora respectively.
The study provides insights into the vaginal microbiome structure of Indian women that will enable us to unravel the microbial biomarkers and explore the prospective candidates for restoring the vaginal microbiota.


Exposure of pregnant and non-pregnant African women to highly toxic environmental compounds


Identification: Reid, Gregor

Credits: None available.

Exposure of pregnant and non-pregnant African women to highly toxic environmental compounds
 
Gregor Reid1,2*, Stephanie L Collins2, Justin B Renaud3, Amy M McMillan1,2, Jacob Walsh3, Nicholas Nduti4, Remco Kort5, Wilbert Sybesma5, Ivan Mukisa6, and Mark W Sumarah3
1Departments of Microbiology & Immunology, and Surgery, Western University, 2Lawson Health Research Institute, 3Agriculture and Agri-Foods Canada, London, Canada, 4Technical University of Kenya, 5Yoba-for-life Uganda, and 6Makerere University, Uganda
 
The reproductive health of women and their babies can be severely affected by exposure to environmental toxins. A report showing 15% of Canadian women with mercury levels sufficiently high to cause neurological damage to the fetus illustrated the extent of the problem. We previously showed that pregnant women in Tanzania are exposed to high levels of mercury and arsenic, likely from consumption of fish from Lake Victoria, and Kenyan children are exposed to mycotoxins, carcinogenic and teratogenic compounds that enter the food chain. In this study, we explored exposure in 149 pregnant and non-pregnant Rwandan women to aflatoxins (AF), deoxynivalenol (DON), fumonisins (F), ochratoxin A (OTA) and zearalenone (ZEA). Using a state-of-the-art LC-MS/MS approach with isotope-labelled standards, we found that plasma AFB1-lysine, a sensitive biomarker for AFB1 exposure, was widely prevalent in 85% of women at a mean concentration of 2.195 ± 1.750 pg/mg albumin. Urinary AFM1, AFB1, and AFG1 were also found in 47%, 8% and 26% of women at concentrations of 97.9 ± 145.1, 9.08 ± 5.70, and 38.4 ± 79.7 pg/mg creatinine. DON, FB1, FB2, OTA and ZEA were detected in the urine of 60%, 30%, 15%, 71% and 42% of individuals in this population, at a mean quantity of 117.5 ± 342.6 ng/mg creatinine, 9.00 ± 18.16, 51.7 ± 39.8, 24.1 ± 31.2 and 42.9 ± 72.6 pg/mg creatinine, respectively. Although the source of contamination remains unknown, AFM1 levels were elevated in women with daily vegetable intake. In addition, pregnant women had greater exposure to AFB1 consistently across each trimester, compared to their non-pregnant counterparts. There was no correlation with vaginal microbiome abundance profiles or Nugent scores, emphasizing that urinary metabolites provide additional information on the health of the mother and fetus. Laboratory studies showed that Lactobacillus rhamnosus Yoba 2012, a probiotic that along with L. rhamnosus GR-1 is now distributed to over 350,000 people in fermented milk and cereal across east Africa further to program we initiated, can degrade aflatoxins. Daily intake of this food offers the potential to reduce adsorption of these carcinogens.


Possible mechanistic drivers of rapid shifts within the vaginal microbiome


Identification: Schiffer, Joshua

Credits: None available.

Possible mechanistic drivers of rapid shifts within the vaginal microbiome
 
Florencia A. T. Boshier1, Sujatha Srinivasan1, Amalia Magaret1,2, Hyunju Son2, Anthony Lopez1, Sean Proll1, David N. Fredricks1,2, Joshua T. Schiffer1,2
1University of Washington, Seattle
2Fred Hutchinson Cancer Research Center, Seattle
 
Dramatic transitions from Lactobacillus predominance to a complex polymicrobial state correspond with development of bacterial vaginosis (BV). We sought to understand how individual species interact to establish this dysbiotic state. We enrolled 20 women with recurrent BV who self-collected vaginal swabs every 8 hours for 60 days. Swabs were processed for quantitative PCR (qPCR) of seven key species, with daily assessment of bacterial diversity using broad-range PCR and deep sequencing. We generated univariate Poisson mixed models: independent variables were menses, vaginal sex and oral sex; outcome was proportion of days with an increase in bacterial quantity of >0.5 log; time windows between exposure and outcome varied between 0 and 5 days. We mapped bacterial interaction networks using statistical and dynamical systems approaches including Granger causality, LIMITS (Learning Interactions from Microbial Time Series) and SSMAP (Equation Free Modeling Linear Regression Analysis). Three women had stable high levels of Lactobacillus species with periodic introduction but rapid elimination of BV-associated species; five women had high levels of Lactobacillus species with moderate levels of fluctuating BV-associated species; two women demonstrated BV-associated species predominance, high diversity and periodic introduction and elimination of Lactobacillus species; ten women had occasional profound shifts between Lactobacillus predominance and a diverse polymicrobial profile. Shifts of 2-3 logs in individual species occurred in 8-24 hour-windows resulting in complete reorganization of the vaginal microbiome. Menses and sexual activity did not predict shifts in any species. The two dynamical approaches (LIMITS and SSMAP) detected interactions between species that were relatively sparse with evidence of intra- and inter-species competition which varied between individuals, but no growth synergy between pairs.  While the vaginal microbiome is often volatile over intervals of hours in women with BV, the key mechanistic drivers of abrupt shifts in vaginal microbial diversity remain unknown. 
 
This work was funded by the NIH STI CRC (U19 AI 113173)
 

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