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Articles

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
 

 


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.


Acceptability of Vaginal Probiotics Use To Prevent Bacterial Vaginosis Recurrence In High-Risk Rwandan Women


Identification: van de Wijgert, Janneke

Credits: None available.

Acceptability of Vaginal Probiotics Use To Prevent Bacterial Vaginosis Recurrence In High-Risk Rwandan Women
 
Verwijs MC1, Agaba SK2, Uwineza M2, Umulisa MM2, van de Wijgert JHHM2,3
1Institute of Infection and Global Health, University of Liverpool, Liverpool, United Kingdom; 2Rinda Ubuzima, Kigali, Rwanda. 3Julius Center for Health Sciences and Primary Care, University Medical Center Utrecht, Utrecht, The Netherlands
 
Aim: Bacterial vaginosis (BV) in Rwandan women at risk of HIV and sexually transmitted infections is common. We conducted a pilot study to determine the feasibility, acceptability, and preliminary efficacy of three interventions to prevent BV recurrence.
 
Methods: 68 women who completed oral metronidazole treatment for BV and/or Trichomonas vaginalis were randomised to four groups (N=17 each): no product, or intermittent use of oral metronidazole, or one of two vaginal probiotics. Participants inserted the first dose under direct observation and used products for two months. They were counselled to practice safer sex, cease vaginal practices, and encourage male partner penile hygiene. Data were collected in face-to-face and in-depth interviews, daily diaries, and focus group discussions.
 
Results: Most women (93%) were sex workers and the BV prevalence (by Nugent) at baseline was 83.6%. None had ever heard of or used probiotics. Women were able to insert correctly without practice (100%), inserted before going to sleep (100%), while lying down (94%), and reported that inserting became easier over time (100%). Most women (90%) reported to have used >80% of the required doses (Fisher's exact p=0.371 between groups), and 60% of women had perfect adherence. Most frequently reported reasons of non-adherence were 'simply forgetting' (n=11), being away from home and forgetting the product (n=2), side-effects (n=2), and having menses (n=2). Qualitative data suggested that women believed the products were helpful, but that male partners were not always supportive. 51% of the women at baseline reported to wash inside the vagina but this decreased to 19% during follow-up (McNemar's p<0.001). There were no changes in sexual behaviours (e.g. increased condom use) over time.
 
Conclusion: Vaginal probiotic use is feasible in this setting, and acceptability and adherence were high. Targeted counselling is needed to stimulate safer sex, and acceptability of investigational products by loved ones.
 


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.
 


Factors Influencing Vaginal Practices among Sexually Active Women in an urban Town along a Trans African highway in Southwestern Uganda


Identification: Bayigga, Josephine

Credits: None available.

Factors Influencing Vaginal Practices among Sexually Active Women in an urban Town along a Trans African highway in Southwestern Uganda
 
Josephine Bayigga 1, 2, Martin John Buwembo1, Christine Atuhairwe 2
1MRC/UVRI &LSHTM Uganda Research Unit, Entebbe
2Clarke International University, Namuwongo, Uganda
 
Background
Vaginal practices are done worldwide by women to attain a projected desirable state. The frequency, application and motivations vary by region and population. Several researchers report vaginal practices as a contributing factor to Sexually Transmitted Infections (STIs). We investigated factors influencing vaginal practices among sexually active women in Lyantonde town and the relationship with STI acquisition.
 
Methods
A cross sectional study was conducted among 143 sexually active women ages 18-45 years from Lyantonde town (Latitude: -0.4070; Longitude: 31.1575) along the Trans African highway in Southwestern Uganda. These were selected through convenience random sampling. Interviewer administered questionnaires and two focus group discussions were conducted to collect data.  Statistical data analysis was done using Stata version 12, Chi-square test was used to determine association and a p-value of 0.05 was considered significant. Qualitative data was thematically analyzed.
 
Results
Almost all women reported vaginal practices (98.6%, n=141) these included use of; water 24% (n=107), douching 26%(n=116),  cloth 20%(n=88), soap 17% (n=78), herbs 11% (n=48), others were  washing powder, perfume, soda, Jelly, cooking oil, cotton and paper at 3%(n=13). General hygiene was the main motivation reported by all of the women, followed by cleaning before or after sex 90.9% (n=130), preparing the vagina for sex (not to clean) 79.7% (n=114) and to treat infections 29.4% (42). Cultural norms and values,  peer influence, the persuasive market and affordability accelerated utilization of vaginal products. Education was associated with use of vaginal practices x2 12.359, p-value 0.015. Lesser educated women were three times likely to engage in vaginal practices compared to their counterparts. There was no association between vaginal practices and susceptibility to STIs among in this population.
 
Conclusion
General hygiene is a major motivation for vaginal practices in this population. However, incidence of STIs could have also been affected by under or over reporting of symptoms.
 

 


Association between dysbiosis of the vaginal microbiota and bacterial infections


Identification: ., Apoorva

Credits: None available.

Association between dysbiosis of the vaginal microbiota and bacterial infections
 
Apoorva1, Somesh Gupta1*, Seema Sood2, Benu Dhawan2, Garima Kacchawa3, A.K.Saxena4
Departments of 1Dermatology and Venereology,2Microbiology, 3Obstetrics and Gynaecology, All India Institute of Medical Sciences, New Delhi, India, 4Dermatology and S.T.D Department, Vardhman Mahavir Medical College and Safdarjung Hospital, New Delhi, India
*Corresponding author
 
Introduction: A healthy vaginal flora is composed of facultative and strict anaerobes with Lactobacillus species being predominant. Decrease in the protective lactobacilli and concomitant increase in anaerobic bacteria leads to bacterial vaginosis (BV). Alterations in the composition of the vaginal flora is associated with adverse reproductive outcomes and acquisition and transmission of sexually transmitted infections (STIs) e.g. Neisseria gonorrhea, Chlamydia trachomatis, Mycoplasma species and human immunodeficiency virus (HIV).
 
Aim & Objectives: The study was conducted to assess the prevalence of bacterial infections of vaginal tract in women with abnormal flora and its correlation with severity of dysbiosis.
MATERIALS & METHODS: Thirty five sexually active non-pregnant women between 18-45 years of age who presented with self reported symptoms of vaginitis (vaginal discharge/ genital itching/ genital burning/ lower abdominal pain) were included in the study. Vaginal swab specimens were collected from the lateral wall of the vagina and abundance of various bacterial morphotypes was assessed by microscopic evaluation of Gram stained smears using the Nugent scoring method for diagnosis of BV. Samples with a score of ≥4 suggestive of dysbiosis were processed further for polymerase chain reaction (PCR) based detection of Neisseria gonorrhea, Chlamydia trachomatis, Mycoplasma genitalium, Mycoplasma hominis and Ureaplasma urealyticum.
 
Results: Nugent's score ≥4 was reported in 15 (42.86%) out of 35 women enrolled in the study. Among these women with dysbiosis, Chlamydia trachomatis was detected in 1 (6.67%), Mycoplasma hominis in 2 (13.33%) and Ureaplasma urealyticum in 1 (6.6.7%) out of 15 samples. None of these 15 samples were positive for Neisseria gonorrhea and Mycoplasma genitalium. Co-infection with other bacteria was found in BV positive women.
 
Conclusion: Vaginal dysbiosis may be associated with other bacterial infections and therefore regular screening for co-existing pathogens contributing to STIs should be encouraged.
 

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