Poster Abstracts - S618
Articles
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.
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
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.
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.
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.
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
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)
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)
Exploring microbe-immune crosstalk: immunoglobulin coating of vaginal microbiota
Identification: Schuster, Heleen
Credits: None available.
Exploring microbe-immune crosstalk: immunoglobulin coating of vaginal microbiota
Heleen Schuster MDa,b, Annelot Breedveld MScc, Andries Budding MD PhDa, Paul Savelkoul PhDa,d
aDepartment of Medical Microbiology and Infection Control, Amsterdam UMC, Vrije Universiteit Amsterdam, Amsterdam, Netherlands; bDepartment of Obstetrics and Gynecology, Amsterdam UMC, Univ of Amsterdam, Amsterdam, Netherlands; cDepartment of Molecular Cell Biology and Immunology, Amsterdam UMC, Vrije Universiteit Amsterdam, Amsterdam, Netherlands; dDepartment of Medical Microbiology, Maastricht University Medical Centre, Maastricht, Netherlands
As knowledge on the role and composition of the vaginal microbiome is accumulating, more interest is directed to microbe:immune crosstalk. Local immunoglobulins (Igs) play an important role as mediator of mucosal immunity and homeostasis. In the vaginal mucosa, IgG is the dominating antibody, while at other mucosal surfaces IgA is most prevalent. In the gut, alterations in IgA coating of commensal bacteria are associated with inflammatory bowel disease and Clostridium difficile infection. Less is known about the role of local Igs in the female genital tract. We aim to provide insight in the IgA and IgG coating of vaginal bacteria using a combination of flow cytometry based techniques and microbiota profiling.
Vaginal swabs were collected from healthy volunteers. Fluorescence-Activated Cell Sorting (FACS) was used to sort micro-organisms according to IgA and IgG coating. The bacterial composition was analyzed with bacterial profiling technique IS-pro, using fragment variability of the 16S-2S rDNA intergenic spacer region.
Lactobacillus crispatus and L. iners dominated the vaginal microbiota of healthy volunteers and show large numbers of IgA and IgA/IgG coated Lactobacilli. Bacteria with only IgG on their surface were not detected. The proportions of Ig coated bacteria varied among individuals with similar microbial compositions.
Vaginal bacteria show varying amounts of IgA and IgA/IgG coating between different healthy volunteers. The high amount of Ig coated vaginal commensal bacteria demonstrate the importance of local Igs in the mucosal environment. Disturbances of the coating might lead to increased local inflammation and can possibly identify women at risk for diseases of the reproductive tract and adverse pregnancy outcomes. These promising results indicate a putative role of Igs in the maintenance of a healthy microbiome.
Heleen Schuster MDa,b, Annelot Breedveld MScc, Andries Budding MD PhDa, Paul Savelkoul PhDa,d
aDepartment of Medical Microbiology and Infection Control, Amsterdam UMC, Vrije Universiteit Amsterdam, Amsterdam, Netherlands; bDepartment of Obstetrics and Gynecology, Amsterdam UMC, Univ of Amsterdam, Amsterdam, Netherlands; cDepartment of Molecular Cell Biology and Immunology, Amsterdam UMC, Vrije Universiteit Amsterdam, Amsterdam, Netherlands; dDepartment of Medical Microbiology, Maastricht University Medical Centre, Maastricht, Netherlands
As knowledge on the role and composition of the vaginal microbiome is accumulating, more interest is directed to microbe:immune crosstalk. Local immunoglobulins (Igs) play an important role as mediator of mucosal immunity and homeostasis. In the vaginal mucosa, IgG is the dominating antibody, while at other mucosal surfaces IgA is most prevalent. In the gut, alterations in IgA coating of commensal bacteria are associated with inflammatory bowel disease and Clostridium difficile infection. Less is known about the role of local Igs in the female genital tract. We aim to provide insight in the IgA and IgG coating of vaginal bacteria using a combination of flow cytometry based techniques and microbiota profiling.
Vaginal swabs were collected from healthy volunteers. Fluorescence-Activated Cell Sorting (FACS) was used to sort micro-organisms according to IgA and IgG coating. The bacterial composition was analyzed with bacterial profiling technique IS-pro, using fragment variability of the 16S-2S rDNA intergenic spacer region.
Lactobacillus crispatus and L. iners dominated the vaginal microbiota of healthy volunteers and show large numbers of IgA and IgA/IgG coated Lactobacilli. Bacteria with only IgG on their surface were not detected. The proportions of Ig coated bacteria varied among individuals with similar microbial compositions.
Vaginal bacteria show varying amounts of IgA and IgA/IgG coating between different healthy volunteers. The high amount of Ig coated vaginal commensal bacteria demonstrate the importance of local Igs in the mucosal environment. Disturbances of the coating might lead to increased local inflammation and can possibly identify women at risk for diseases of the reproductive tract and adverse pregnancy outcomes. These promising results indicate a putative role of Igs in the maintenance of a healthy microbiome.
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
Impacts of microbial derived short chain fatty acids in regulating immune activation at the female reproductive tract via epigenetic mechanisms
Identification: Siddik, Abu Bakar
Credits: None available.
Impacts of microbial derived short chain fatty acids in regulating immune activation at the female reproductive tract via epigenetic mechanisms
Abu Bakar Siddik1, Chih-Yu Chen2, Chris Grant2, Garrett Westmacott2, T. Blake Ball1,2, Ruey-Chyi Su1,2,*
1University of Manitoba, Medical Microbiology & Infectious Diseases; 2National HIV and Retrovirology Laboratory, JC Wilt Infectious Diseases Research Centre, Winnipeg, Manitoba, Canada
*Corresponding author.
Epithelia are physical barriers, as well as key regulators of mucosal immunity. Soluble mucosal factors such as short chain fatty acids (SCFAs) in the vaginal fluids (VF) can alter the epigenetic regulation of immunologic genes via inhibition of cellular histone deacetylase (HDAC) activities. The role of HDAC is to deacetylase histone which consequently wrapped the DNA tightly with the histone core. So the inhibition of HDAC cause keeping DNA in open conformation which eventually become favourable for gene expression. This study explores the impact of SCFAs in regulating the transcription of immunologic genes in epithelial cells derived from the female reproductive tract (FRT). To define the effects of sodium butyrate (NaB), one of the vaginal SCFAs, on the expression of anti- and pro-inflammatory mediators, and to identify cellular proteins that are specifically affected by exposure to NaB. The vaginal epithelial cell line 'Vk2' was pre-treated with NaB at non-toxic concentrations prior to stimulation with Toll-like-receptor (TLR) agonists. Cellular RNA was quantitated with RT-qPCR, and cellular proteins were analyzed for post-translational modifications (i.e. acetylation, methylation, and phosphorylation) using proteomic profiling. NaB (5mM) significantly enhanced transcription of the pro-inflammatory cytokines IL-6 and TNFα, and only TNFα transcript levels rose in response to TLR-1, -2, -3, -5, and -6 stimulation. A change in the profile of phosphorylated proteins was also observed following NaB-treatment in the absence of TLR stimulation. Proteins that lost phosphorylation in NaB-treated Vk2 cells were implicated in cell-cell adhesion and mRNA splicing. This suggests that changes in cytokine/chemokine expression may be affected at transcription or post-transcription. In addition, NaB may affect epithelial barrier function by altering protein phosphorylation.
Abu Bakar Siddik1, Chih-Yu Chen2, Chris Grant2, Garrett Westmacott2, T. Blake Ball1,2, Ruey-Chyi Su1,2,*
1University of Manitoba, Medical Microbiology & Infectious Diseases; 2National HIV and Retrovirology Laboratory, JC Wilt Infectious Diseases Research Centre, Winnipeg, Manitoba, Canada
*Corresponding author.
Epithelia are physical barriers, as well as key regulators of mucosal immunity. Soluble mucosal factors such as short chain fatty acids (SCFAs) in the vaginal fluids (VF) can alter the epigenetic regulation of immunologic genes via inhibition of cellular histone deacetylase (HDAC) activities. The role of HDAC is to deacetylase histone which consequently wrapped the DNA tightly with the histone core. So the inhibition of HDAC cause keeping DNA in open conformation which eventually become favourable for gene expression. This study explores the impact of SCFAs in regulating the transcription of immunologic genes in epithelial cells derived from the female reproductive tract (FRT). To define the effects of sodium butyrate (NaB), one of the vaginal SCFAs, on the expression of anti- and pro-inflammatory mediators, and to identify cellular proteins that are specifically affected by exposure to NaB. The vaginal epithelial cell line 'Vk2' was pre-treated with NaB at non-toxic concentrations prior to stimulation with Toll-like-receptor (TLR) agonists. Cellular RNA was quantitated with RT-qPCR, and cellular proteins were analyzed for post-translational modifications (i.e. acetylation, methylation, and phosphorylation) using proteomic profiling. NaB (5mM) significantly enhanced transcription of the pro-inflammatory cytokines IL-6 and TNFα, and only TNFα transcript levels rose in response to TLR-1, -2, -3, -5, and -6 stimulation. A change in the profile of phosphorylated proteins was also observed following NaB-treatment in the absence of TLR stimulation. Proteins that lost phosphorylation in NaB-treated Vk2 cells were implicated in cell-cell adhesion and mRNA splicing. This suggests that changes in cytokine/chemokine expression may be affected at transcription or post-transcription. In addition, NaB may affect epithelial barrier function by altering protein phosphorylation.
Morphotypic variation of vaginal clue cells broadens understanding of vaginal biofilms
Identification: Sycuro, Laura
Credits: None available.
Morphotypic variation of vaginal clue cells broadens understanding of vaginal biofilms
Laura K. Sycuro*,1,2, Shaelen Konschuh1, George Kuo2, and David N. Fredricks2,3
1International Microbiome Centre, University of Calgary; 2Fred Hutchinson Cancer Research Center; 3University of Washington
*Corresponding author
Bacterial vaginosis (BV) is a dysbiotic state of the vaginal microbiota that affects 10-60% of premenopausal women globally. BV increases a woman's risk of acquiring sexually transmitted infections (STI) and experiencing pregnancy complications such as preterm birth. One of the pivotal problems in the clinical management of BV is its high rate of recurrence. A primary hypothesis for why BV recurs is the presence of an adherent vaginal biofilm that enables microbes to survive exposure to antimicrobials. Here we report a longitudinal sub-study aimed at describing the morphotypic appearance of vaginal epithelial cells coated in adherent biofilm (clue cells) from diagnosis and treatment through 3-6 months of follow-up. Of 85 mostly African American (58%) and White (34%) women enrolled through the Public Health Seattle and King County Sexually Transmitted Diseases Clinic, 38 (45%) completed all 6 months of follow-up and 60% experienced BV recurrence. Most participants (73/74) who were BV-positive at the baseline visit had vaginal clue cells observed in their vaginal fluid. Freshly collected cervicovaginal lavage fluid from the study's 374 patient visits was microscopically observed, Gram stained, and fixed for downstream analysis. Using microscopy and molecular probing we observed that in addition to the established clue cell morphotype consisting predominantly of Gardnerella vaginalis and Atopobium vaginae, there was a second, although rare morphotype dominated by curved rod bacteria. We confirmed the identity of the abundantly adherent bacteria in some samples was Mobiluncus mulieris, and subsequent in vitro studies suggested this species' ability to form adherent biofilms is strain-variable. To our knowledge, this study is the first to report bacteria other than G. vaginalis and A. vaginae can dominate the vaginal biofilm and contribute a distinct morphotypic variation of vaginal clue cells. Efforts to identify the other curved rod bacteria contributing to the 'fuzzy' appearance of vaginal clue cells are on-going, as is a systematic assessment of how variable clue cell morphotypes are linked to patient outcomes.
Laura K. Sycuro*,1,2, Shaelen Konschuh1, George Kuo2, and David N. Fredricks2,3
1International Microbiome Centre, University of Calgary; 2Fred Hutchinson Cancer Research Center; 3University of Washington
*Corresponding author
Bacterial vaginosis (BV) is a dysbiotic state of the vaginal microbiota that affects 10-60% of premenopausal women globally. BV increases a woman's risk of acquiring sexually transmitted infections (STI) and experiencing pregnancy complications such as preterm birth. One of the pivotal problems in the clinical management of BV is its high rate of recurrence. A primary hypothesis for why BV recurs is the presence of an adherent vaginal biofilm that enables microbes to survive exposure to antimicrobials. Here we report a longitudinal sub-study aimed at describing the morphotypic appearance of vaginal epithelial cells coated in adherent biofilm (clue cells) from diagnosis and treatment through 3-6 months of follow-up. Of 85 mostly African American (58%) and White (34%) women enrolled through the Public Health Seattle and King County Sexually Transmitted Diseases Clinic, 38 (45%) completed all 6 months of follow-up and 60% experienced BV recurrence. Most participants (73/74) who were BV-positive at the baseline visit had vaginal clue cells observed in their vaginal fluid. Freshly collected cervicovaginal lavage fluid from the study's 374 patient visits was microscopically observed, Gram stained, and fixed for downstream analysis. Using microscopy and molecular probing we observed that in addition to the established clue cell morphotype consisting predominantly of Gardnerella vaginalis and Atopobium vaginae, there was a second, although rare morphotype dominated by curved rod bacteria. We confirmed the identity of the abundantly adherent bacteria in some samples was Mobiluncus mulieris, and subsequent in vitro studies suggested this species' ability to form adherent biofilms is strain-variable. To our knowledge, this study is the first to report bacteria other than G. vaginalis and A. vaginae can dominate the vaginal biofilm and contribute a distinct morphotypic variation of vaginal clue cells. Efforts to identify the other curved rod bacteria contributing to the 'fuzzy' appearance of vaginal clue cells are on-going, as is a systematic assessment of how variable clue cell morphotypes are linked to patient outcomes.