Poster Abstracts - S618


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.

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.

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)

The vaginal microbiota of HIV –infected pregnant women: associations with local inflammation and gestational age at delivery

Identification: Short, Charlotte

Credits: None available.


The vaginal microbiota of HIV -infected pregnant women: associations with local inflammation and gestational age at delivery
C Short1, R Quinlan1, R Brown1, Y Lee1, R Shattock1, P Bennett1, G Taylor1, D MacIntyre1 and London HIV Pregnancy Research Group
1Imperial College London
HIV + women experience high rates of PTB, in spite this, there is little data on their vaginal microbiota during pregnancy. We sought to describe the microbiome in a group of HIV+ pregnant women and explore associations with local cytokine environment and gestational age at delivery (GA).
A prospective observational multi-site study. Vaginal and cervicovaginal fluid (CVF) were obtained using a swab and menstrual softcup during the 2nd trimester from HIV+ and HIV- pregnant women (Exclusion criteria: <350 cells/mm3, multiple or in-vitro pregnancy and injecting drug user). MiSeq sequencing of 16S rRNA gene amplicons was used to characterize the vaginal microbiome. Multiplex assays were used to measure CVF cytokine concentrations. Multivariate modeling was performed to explore associations with bacterial genus/species, CVF cytokine concentrations and clinical data.
HIV+ women (n=53) had a median age of 35, 85% were Black and 14% had PTB. HIV- women (n=30) had a median age of 33 and 50% were Caucasian. HIV+ women delivering at term had higher abundance of Gardnerella (18% versus 3% p=0.003) and Prevotella genera (4% versus 0.1% p=0.002) and lower proportions of Lactobacillus species (70% versus 93% p=0.009) compared to HIV- women. The predominant vaginal community state type (CST) of HIV+ pregnant women was III (L.iners dominant) 55% (n=29), 26% (14) were CST IV (high diversity, anaerobic), 13% (7) were CST I and 2% (1) were CST II. Amongst HIV+ women, PTB was associated with increased proportions of Gardnerella spp. (p<0.0001), Prevotella spp. (p<0.0001), Aerococcus spp. (p=0.015) and Megasphaera spp. (p=0.03) compared with term birth.  All PTBs were in women assigned to CST III and IV. The proportion of read counts of CST IV associated anaerobes were positively correlated with CVF IFNγ, IL-1β, IL12p70 and TNFα. IL-1β was positively associated with bacterial diversity and richness. No associations between cytokine concentrations and GA were observed.
Diverse vaginal bacterial communities in HIV+ women are associated with PTB. The associated local proinflammatory cytokine profile may reflect the pathogenic contribution of these organisms to the early trigger of labor.
Funding: Wellcome Trust


Networks of Co-Abundant Bacteria that Increase Prior to Incident Bacterial Vaginosis

Identification: Taylor, Christopher

Credits: None available.

Networks of Co-Abundant Bacteria that Increase Prior to Incident Bacterial Vaginosis
Christopher M Taylor1, Omar Aldibasi1,2, Vincent Maffei1, Eugene Blanchard IV1, David H Martin1,3, Jane R Schwebke4, Christina A Muzny4
1Louisiana State University Health Sciences Center; New Orleans, LA, USA; 2King Abdullah International Medical Research Center; Riyadh, Saudi Arabia; 3Tulane University; New Orleans, LA, USA; 4University of Alabama at Birmingham; Birmingham, AL, USA      
Bacterial vaginosis (BV) is the most common vaginal infection and its etiology remains unknown. This study aims to determine co-abundant vaginal bacteria leading up to incident BV (Nugent score of 7-10 for at least 2 consecutive days). A cohort of African American women who have sex with women, with normal vaginal flora (Nugent 0-3 and no Amsel criteria), were prospectively enrolled and followed for 90 days with daily self-collected vaginal swabs to determine the sequence of microbiological events occurring prior to incident BV (iBV) [a]. We analyzed 176 vaginal specimens from 14 participants who developed iBV during the study using SParse InversE Covariance Estimation for Ecological ASsociation Inference (SPIEC-EASI) [b] in order to assess microbial networks arising in the 14 days leading up to iBV. A robust network of 3 bacteria was identified, consisting of Gardnerella vaginalis, Atopobium vaginae, and Aerococcus christensenii, which increased in abundance prior to iBV (p-value < 0.01). Aerococcus christensenii is particularly interesting due to its role as a pathogen in the female genital tract [c] and involvement in a case of severe polymicrobial chorioamnionitis [d], likely due to an ascending vaginal infection.
a) Muzny CA, Blanchard E, Taylor CM, et al. Identification of Key Bacteria Involved in the Induction of Incident Bacterial Vaginosis: A Prospective Study. J Infec Dis. 2018 Apr
b) Kurtz ZD, Muller CL, Miraldi ER, et al. Sparse and compositionally robust inference of microbial ecological networks. PLoS Comput Biol. 2015 May
c) Rasmussen M.  Aerococcus: an increasingly acknowledged human pathogen. Clin Microbiol Infect. 2016 Jan
d) Carlstein C, Marie Soes L, Jorgen Christensen J. Aerococcus christensenii as Part of Severe Polymicrobial Chorioamnionitis in a Pregnant Woman. Open Microbiol J. 2016 Mar
Funding is provided through NIH awards K23 AI106957 to CAM and R01 AI118860 to CMT.
Funding was also provided through NIH awards U54 TR001368 and P20 GM103424 for CMT.

Ubiquitous gram-positive bacteria in the male urogenital tract are disease causing agents in chronic pelvic pain

Identification: Thumbikat, Praveen

Credits: None available.

Ubiquitous gram-positive bacteria in the male urogenital tract are disease causing agents in chronic pelvic pain
Praveen Thumbikat1, Stephen F. Murphy1, Jonathan F. Anker1, Daniel J. Mazur1, Anthony J. Schaeffer1, Christel Hall1
1Feinberg School of Medicine, Northwestern University, Chicago, IL 60611
Gram-positive bacteria are considered an ubiquitous but incidental finding in urine and prostatic secretion cultures from male patients with chronic pelvic pain syndrome (CPPS)  However, we hypothesized that gram-positive traditional non-uropathogenic bacteria can play a direct role in CPPS pathogenesis. Staphylococcus epidermidis, Staphylococcus haemolyticus, and Enterococcus faecalis comprised 62% of prostate-localizing bacteria from CPPS patients. A subset of these induced high NFκB expression, contained pathogenic virulence factors, and were associated with increased clinical symptoms. Here we show that an immunogenic subset of the most common gram-positive prostate-localizing bacterial strains can directly cause pelvic tactile allodynia and voiding dysfunction. Intra-urethral infection of mice with immunogenic bacterial isolates from each strain, but not an S. epidermidis isolate from a healthy control, recapitulated symptoms seen in patients. Mechanistically, these pathogenic gram-positive bacteria induced unique prostate inflammation, involving T cells, NK cells, and cytokine production. We demonstrate for the first time that the prevalent, yet untreated traditional non-uropathogenic gram-positive bacteria can play a causative role in CPPS pathogenesis.

Role of Bacterial Vaginosis associated bacterial sialidase in HIV infection

Identification: Traviss, Riley

Credits: None available.

Role of Bacterial Vaginosis associated bacterial sialidase in HIV infection
Riley Traviss and Zenda Woodman
University of Cape Town, Integrative Biomedical Sciences, Cape Town, South Africa
Studies suggest that women with Bacterial Vaginosis (BV) are at a higher risk of being infected by HIV-1 than those with a microbiome dominated by Lactobacillus spp.. A potential mechanism is the secretion of sialidase by BV-associated bacteria (BVAB) that modify either host cell and/or viral N-glycans, facilitating binding of Envelope (Env) to CD4 and/or CCR5 and thus enhance viral entry.  When TZM-bl cells were infected with pseudovirus in the presence of sialidase, infection increased 2-fold and this effect was reduced upon addition of N-Acetyl-2,3-dehydro-2-deoxyneuraminic acid (DMM), an inhibitor of sialidase. Therefore, the addition of bacterial sialidase enhanced HIV infection. When TZM-bl cells were infected with pseudovirus in the presence of brain heart infusion (BHI) medium used to culture the BVAB, Gardnerella vaginalis (GV), abiotic GV culture medium inhibited pseudovirus entry without affecting TZM-bl cell viability. This suggested that either sialidase was not secreted by the GV strain and/or another factor was influencing pseudovirus infection. When the abiotic BHI medium was adjusted to pH 7.48, the inhibitory effect of GV was reduced 2-fold and when the abiotic BHI medium was boiled, GV culture no longer inhibited pseudovirus entry.  Therefore, GV secreted a heat labile factor that inhibited pseudovirus infection without being toxic to TZM-bl cells. GV secretes Vaginolysin, a member of the cholesterol-dependent cytolysin family believed to contribute to BV pathogenesis. As the MTT test indicated that TZM-bl cells remained viable during incubation with abiotic GV culture medium, they are unlikely to have lysed.  Although we cannot discount the effect of Vaginolysin in our assay, it is possible that anaerobic culture of GV in BHI might stimulate the secretion of a compound that inhibits HIV-1 infection in vivo and that other mitigating factors such as inflammation could negate the protective effect.

Modulation of dendritic cell immune responses by Envelope, and not functional fitness, might be important for subtype C HIV-1 transmission

Identification: Woodman, Zenda

Credits: None available.

Modulation of dendritic cell immune responses by Envelope, and not functional fitness, might be important for subtype C HIV-1 transmission
Evelyn Ngwa Lumngwena1,2, Bahiah Meyer3, Riley Traviss3 and Zenda Woodman3
1Institute for Medical Research and Medicinal Plants Research, MINRESI, Cameroon; 2Faculty of Health Sciences, University of Cape Town, South Africa
HIV-1 Envelope (Env) plays a major role in the competitive ability of the virus and a number of studies have investigated the role that it might play in transmission. One such study identified subtype B transmission motifs: a Histidine at position 12 (H12) of the signal peptide and the absence of a potential N-glycosylation site (PNG) at position 413 (N413), suggested to enhance viral fitness and/or facilitate escape from immune responses. We determined how Env might contribute to the selective advantage of subtype C transmitted founders (TF) by: 1) comparing four TF Env to matched chronic infection (CI) variants, and 2) characterising the impact of H12 and N413 on Env processing and phenotype. All nine subtype C Envs carried a Glutamic acid at position 12 (Q12) and a PNG at 413 representative of the population frequency, suggesting that subtype B TFs might have evolved by a pathway distinct from other subtypes. When a subtype C TF Env was mutated, pseudovirus (PSV) infectivity of constructs Q12H and Q12A decreased significantly, suggesting that H12 was not linked to enhanced infectivity as reported for subtype B TF. On the contrary, deletion of N413 significantly increased PSV entry efficiency, suggesting that even though the presence of a PNG at this site had a fitness cost, it was conserved in subtype C variants, including TFs. Furthermore, the processing and phenotype of TF Envs and PSV entry efficiency did not differ significantly from matched CI variants supporting the suggestion that Env functional fitness was not important for subtype C transmission. However, TFs were better able to stimulate monocyte derived dendritic cells (MDDCs) to secrete higher levels of IL-10, TNF-α, IL-6, IL-8 and MIP1β than their matched CI counterparts. This suggests that subtype C TFs might have the ability to induce immune responses in the female genital tract that favour transmission. Overall, HIV-1 subtype C transmission might not be dependent on Env function but rely more on its ability to induce dendritic cell-mediated immune responses that promote HIV survival within the female genital tract.