Thesis (Ph.D.)--Virginia Commonwealth University, 2010. / Prepared for: Dept. of Microbiology and Immunology. Title from resource description page. Includes bibliographical references.

Worldwide, bacterial vaginosis (BV) is the most common vaginal disorder in women of childbearing age. BV is of major clinical importance due to its ability to significantly affect pregnancy outcome and enhance the transmission and acquisition of HIV. BV is characterized by a dramatic shift in the vaginal microflora; in most BV cases, the predominant bacterial species is Gardnerella vaginalis. It has been demonstrated that G. vaginalis forms an adherent biofilm on the vaginal epithelium of women with BV. Furthemore, evidence suggests that the high rate of recurrence associated with BV is related to incomplete eradication of the biofilm. The overall goal of this study was to characterize G. vaginalis virulence properties, including biofilm formation, in order to better understand the pathogenesis of BV and to improve available treatment methods. In an effort to tease apart the uncertain etiology of this disorder, we utilized in vitro assays to compare three virulence properties of G. vaginalis relative to other BV-associated anaerobes. Only G. vaginalis demonstrated all three virulence properties, including robust biofilm formation. It has been shown that the biofilm phenotype allows its constituent bacteria to be resistant to many negative environmental stimuli. Therefore, we studied the susceptibilities of biofilm vs. planktonic cultures to H2O2 and lactic acid. Biofilms tolerated higher concentrations of both chemicals; however, when the biofilm was proteolytically disrupted, sensitivity to the chemicals returned to planktonic levels. Since our data suggested a critical role for a protein in biofilm formation, a partial genome sequence of G. vaginalis was searched for sequence homology to known biofilm adhesins using the tBLASTn program. This revealed an open-reading frame encoding a hypothetical protein with significant homology to the staphylococcal Bap protein. Antibody towards a portion of the identified gene product was produced in rabbits by inoculation of a recombinant peptide to an antigenic region of the protein. Antibody inhibition assays against biofilm formation, adherence, initial adherence and aggregation were conducted. Relative expression levels of the biofilm-associated protein were analyzed under different conditions by western blot analysis. Finally, the protein was expressed in heterologous hosts and analyzed for an increase in biofilm formation.

Orientador: Camila Marconi / Resumo: A microbiota vaginal normal é predominantemente composta por espécies de Lactobacillus spp., enquanto que o principal tipo de alteração de microbiota vaginal, a vaginose bacteriana (VB), é composta por uma diversidade de bactérias, não pela predominância de uma única espécie. Todavia, dentre as bactérias mais prevalentes nessa condição estão Atopobium vaginae e Gardnerella vaginalis, espécies conhecidamente produtoras de fatores de virulência, dentre eles a capacidade de formar biofilmes vaginais, dificultando a ação de drogas antimicrobianas, como o metronidazol, para o tratamento da VB. No caso particular da G. vaginalis deve-se destacar a produção de sialidases por algumas linhagens que conferem a capacidade de comprometer a barreira mucosa e degradar IgA vaginal, contribuindo para a adesão e proliferação de diversas espécies bacterianas que pertencem ao core patológico da VB. Entretanto, a ação das sialidases sobre a composição desse microbioma vaginal ainda não foi investigada. Dessa forma, os objetivos desse estudo foram: (1) determinar se as cargas cérvico-vaginais das espécies A. vaginae e G. vaginalis na VB estão associadas com o desfecho do tratamento, utilizando o tratamento convencional com metronidazol; (2) avaliar a influência da presença do gene da sialidase de G. vaginalis e da produção de sialidases sobre a composição do microbioma vaginal. Para tanto, foram incluídas 245 mulheres em idade reprodutiva no primeiro estudo, classificadas de acordo com o padrão d... (Resumo completo, clicar acesso eletrônico abaixo) / Doutor

A Chlamydia trachomatis (Ct) à a bactÃria de transmissÃo sexual mais comum em todo o mundo, apesar de existirem poucos dados sobre este agravo na populaÃÃo brasileira. O objetivo da pesquisa foi determinar a prevalÃncia da infecÃÃo por Ct, avaliando os fatores sÃciodemogrÃficos e achados clÃnicos, colposcÃpicos e citopatolÃgicos associados à ocorrÃncia desta infecÃÃo em mulheres jovens na periferia de Fortaleza. Foi realizado um estudo de corte transversal em 200 mulheres sexualmente ativas, com idade entre 12 e 25 anos, atendidas no perÃodo de agosto de 2011 a agosto de 2012, no ambulatÃrio de ginecologia geral do Hospital Distrital Gonzaga Mota â Barra do CearÃ. InformaÃÃes pessoais e dados do exame ginecolÃgico foram anotados em um questionÃrio e as pacientes submeteram-se à coleta de material da endocÃrvice para teste de captura hÃbrida II para Chlamydia trachomatis e para citologia oncÃtica convencional, seguido de colposcopia. Os dados foram analisados utilizando o software Graphpad Prism 5.0, procedendo-se a anÃlise descritiva e analÃtica utilizando o teste t de Student para as variÃveis nominais e o teste exato de Fisher para as variÃveis quantitativas. A prevalÃncia da infecÃÃo por Chlamydia trachomatis foi 15,5% (31/200) em mulheres adolescentes e adultas jovens. NÃo foi encontrada associaÃÃo entre a infecÃÃo estudada e as caracterÃsticas sÃciodemogrÃficas, hÃbitos sexuais, sinais e sintomas questionados, ectopia cilÃndrica e alteraÃÃes colposcÃpicas. Dentre as atipias citolÃgicas, o ASC-US esteve presente em 20,7% dos casos e 4,5% dos controles (p=0,0067, RR=3,452, IC=1,72-6,89), mostrando uma associaÃÃo positiva com a infecÃÃo clamidiana. A G. vaginalis foi encontrada em 54.8% das pacientes infectadas e em 30,7% das pacientes negativas (p=0,0133, RR=2,305. IC=1,21-4,39), mostrando uma relaÃÃo com a infecÃÃo estudada. Concluiu-se que a infecÃÃo por C. trachomatis teve uma prevalÃncia alta na populaÃÃo estudada, que nÃo houve associaÃÃo com fatores de risco sÃcio-demogrÃfico, biolÃgico, com achados clÃnicos e/ou colposcÃpicos. Houve associaÃÃo de ASC-US e G. vaginalis na citologia oncÃtica com a infecÃÃo estudada. / Chlamydia trachomatis (Ct) is the most common bacterial sexually transmitted infection worldwide, but there are few published dates about it in Brazil. The aim of this study was to determinate the prevalence of Chlamydia trachomatis infection and to assess the socialdemographic behavioral, clinical and cytopathological factors associated with this infection among adolescents and young women in a low-income area of Fortaleza - Brazil. A cross-sectional study was conducted in 200 sexually active women aged 12 to 25 years, from august 2011 to august 2012, in gynecology outpatient clinic of the General Hospital District Gonzaga Mota - Barra do CearÃ. Personal information and date of gynecogical examination were recorded in a questionnaire and the patients underwent collection of material from the endocervix to hybrid capture test for C. trachomatis and Pap test, followed by colposcopy. Data were analyzed using Graphpad Prism 5.0 software, proceeding to descriptive and analytical analysis using the Student t test for nominal variables and the Fisher exact test for quantitative variables. No association was found between infection and studied the socio-demographic characteristics, sexual habits, signs and symptoms questioned, cylindrical ectropion and colposcopic changes.Among the abnormal cytological atypia, ASC-US was presented in 20,7% of cases and 4,5% of controls (p=0,0067, RR=3,452, IC=1,72-6,89), with a positive association with the infection. G. vaginalis morphotype was identified in 54,8% of infected women and 30,7% of negative patients (p=0,0133, RR=2,305. IC=1,21-4,39), showing a relationship with the infection. It was concluded that infection with C. trachomatis had a high prevalence in the population studied, no association was observed with socio-demographic, biological and clinical findings and colposcopic changes. There was association of ASC-US and G. vaginalis in cytology with the infection studied.

Made available in DSpace on 2014-06-12T18:28:04Z (GMT). No. of bitstreams: 2 arquivo1143_1.pdf: 1341192 bytes, checksum: 28c1a0abdeb803275cd83cc199fae089 (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2009 / INSTITUTO DE MEDICINA INTEGRAL PROF FERNANDO FIGUEIRA / Esta tese, apresentada em dois artigos, teve por objetivo comparar dois tratamentos para Vaginose Bacteriana, utilizando metronidazol e aroeira, em aplicação tópica vaginal e estudar o perfil clínico, epidemiológico e bacteriológico das mulheres participantes, antes de serem submetidas ao tratamento. O primeiro artigo consistiu de um ensaio clínico randomizado, duplamente mascarado, que comparou a eficácia entre os dois tratamentos em mulheres com vaginose bacteriana diagnosticada, concomitantemente, pelos critérios de Amsel e Nugent. Foi utilizada a Análise por Intenção de Tratar. Do total de 277 participantes do ensaio clínico, 137 mulheres foram tratadas com gel de Aroeira e 140 tratadas com Metronidazol. Na avaliação de cura pelos critérios de Amsel, 21,2% das pacientes que utilizaram aroeira e 62,1% que usaram metronidazol obtiveram cura. Quando o Escore de Nugent foi utilizado foram curadas 13,9% das mulheres do grupo da aroeira e 56,4% do grupo metronidazol. A cura total (com a utilização dos dois critérios) foi observada em 12,4% do total de pacientes no grupo da aroeira e 56,4% das mulheres que usaram metronidazol. O segundo artigo constou de uma série de casos onde foram estudados os achados clínicos, epidemiológicos e microbiológicos das participantes do ensaio clínico antes do tratamento, além da presença de lactobacilos nas citologias oncóticas e a população bacteriana componente das microbiotas vaginais identificadas por culturas de secreção vaginal. Entre as queixas clínicas, as mais frequentes foram o corrimento genital, observado em 74,4% das participantes e o odor de peixe da secreção vaginal, que ocorreu em 68,6% dos casos. As culturas de conteúdo vaginal permitiram a identificação de Gardnerella vaginalis em 96,8% e de Mobiluncus em 53,1% dos casos. Apenas em uma terça parte dos exames (32,1%) havia a presença de Lactobacillus. Como conclusões dos estudos, foram observados índices de cura menores com a utilização de Aroeira do que com Metronidazol; os efeitos colaterais foram pouco frequentes e sem maior gravidade em ambos os grupos e continuam necessários novos estudos que melhor elucidem as inter-relações entre os achados microbiológicos e a expressão clínica da doença

Submitted by Renata Lopes (renatasil82@gmail.com) on 2017-06-21T19:28:58Z No. of bitstreams: 1 danielemariaknuppdesouza.pdf: 8224549 bytes, checksum: bc3a45c095a6504198c18b63f1922033 (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2017-08-07T19:19:03Z (GMT) No. of bitstreams: 1 danielemariaknuppdesouza.pdf: 8224549 bytes, checksum: bc3a45c095a6504198c18b63f1922033 (MD5) / Made available in DSpace on 2017-08-07T19:19:03Z (GMT). No. of bitstreams: 1 danielemariaknuppdesouza.pdf: 8224549 bytes, checksum: bc3a45c095a6504198c18b63f1922033 (MD5) Previous issue date: 2013-04-25 / FAPEMIG - Fundação de Amparo à Pesquisa do Estado de Minas Gerais / Vaginose Bacteriana (VB) é uma síndrome polimicrobiana, caracterizada pelo desequilíbrio da microbiota vaginal, associada à substituição da população bacteriana vaginal predominantemente aeróbia (Lactobacillus spp.) por uma população anaeróbia (principalmente Gardnerella vaginalis), gerando descarga vaginal anormal em mulheres em idade reprodutiva. O objetivo desse trabalho foi a avaliação de aspectos fisiológicos e moleculares de G. vaginalis em pacientes com e sem VB, atendidas na rede pública (SUS) e privada de Juiz de Fora/MG, além da determinação do perfil de susceptibilidade a antimicrobianos. Amostras de secreção vaginal foram coletadas e processadas para isolamento seletivo, conforme descrito na literatura. G. vaginalis foi identificada presuntivamente pela β-hemólise ou hemólise difusa e os testes da oxidase e catalase. A identidade bacteriana foi confirmada por reação de POR. O gene codificador para a vaginolisina (vly) foi detectado por PCR. O perfil de susceptibilidade a drogas antimicrobianas foi determinado pelo método da diluição em ágar, de acordo as recomendações do CLSI para microrganismos anaeróbios. A genotipagem foi realizada por AP-PCR. De 89 pacientes, G. vaginalis foi isolada de 42 por cultivo em meio de cultura, sendo 35 com VB, 02 com quadro intermediário e 05 saudáveis. Em 47 pacientes não foi possível realizar o isolamento de G. vaginalis por método de cultivo, sendo a reação de PCR positiva para 26 destas pacientes. Para classificar as pacientes em sintomáticas e saudáveis, foi realizado o escore de Nugent, Todos os 204 isolados de G. vaginalis submetidos à reação de PCR tiveram sua identidade confirmada, e destes, em 96,5% foi detectado o gene vly. Quanto ao perfil de susceptibilidade aos antimicrobianos, alta sensibilidade a ampicilina/sulbactam, clindamicina e cloranfenicol foi observada, e alta resistência foi observada a ampicilina, metronidazol, secnidazol e tinidazol. A cultura positiva de G. vaginalis, per se, não garante o diagnóstico de VB, já que os microrganismos fazem parte da microbiota residente. Não foi possível correlacionar a detecção do gene vly como marcador para determinação de linhagens patogênicas. A resistência bacteriana frente aos antimicrobianos mais utilizados na clínica corroboram outros estudos, e alertam para os riscos da terapia empírica rotineira. A técnica de genotipagem utilizada não permitiu o agrupamento de G. vaginalis, de acordo com sua origem de isolamento, sugerindo, assim, uma heterogeneidade populacional. Essa observação sustenta-se pelo não agrupamento bacteriano também em função da tipagem pelo perfil de susceptibilidade a antimicrobianos, embora outras técnicas de genotipagem não tenham sido utilizadas. Além disso, ainda não são descritos oligonucleotídeos iniciadores específicos para genotipagem de G. vaginalis por AP-PCR. Espera-se que esses resultados possam servir de base para suscitar discussões relacionadas ao tratamento empírico da vaginose bacteriana, considerando-se limitações na disponibilidade de diagnóstico ou mesmo terapia antimicrobiana. / Bacterial vaginosis (BV) is a polymicrobial syndrome characterized by an imbalance in the vaginal microbiota associated with the replacement of the vaginal bacterial population predominantly aerobic (Lactobacillus spp.) by an anaerobic population (especially Gardnerella vaginalis) leading to abnormal vaginal discharge in women of reproductive age. The aim of this study was to evaluate physiological and molecular aspects of G. vaginalis in patients with and without BV, treated at public (SUS) and private services in Juiz de Fora/MG, in addition to determining the antimicrobial susceptibility patterns. Vaginal secretion samples were collected and processed for selective isolation, as described in the literature. G. vaginalis was presumptively identified by 13-hemolysis or diffuse hemolysis and oxidase and catalase negative tests. The bacterial identity was confirmed by PCR. The encoding gene for vaginolisin (vly) was detected by PCR. Antimicrobial susceptibility patterns were determined by agar dilution method, according to CLSI guidelines for anaerobic microorganisms. Genotyping was performed by AP-PCR. Out of 89 patients, G. vaginalis was isolated from 42 by cultivation in culture medium, 35 of which with VB, 02 with intermediate clinical and 05 healthy. In 47 patients it was not possible to perform the isolation of G. vaginalis by cultivation method, with a positive PCR result for 26 of these patients. To classify patients in symptomatic and healthy, Nugent score was performed. All 204 isolates of G. vaginalis subjected to PCR reaction had its identity confirmed and, of these, vly gene was detected in 96.5%. Regarding antimicrobial susceptibility patterns, high sensitivity to ampicillin/sulbactam, clindamycin and chloramphenicol was observed, and high resistance was observed to ampicillin, metronidazole, secnidazole and tinidazole. The positive culture of G. vaginalis, per se, does not ensure the diagnosis of BV, as the microorganism may be considered resident microbiota. It was not possible to correlate vly gene detection as a marker for determination of pathogenic strains. Bacterial resistance against antimicrobial drugs commonly used in clinical corroborate with others studies, and warn to the risks of empiric therapy routine. The genotyping technique used did not allow G. vaginalis grouping, according to their source of isolation, suggesting, therefore, a heterogeneous population. This observation is sustained by the impossibility of grouping using the bacterial typing by antimicrobial susceptibility profile too, although other genotyping techniques have not been used. In addition, specific primers for genotyping G. vaginalis by AP-PCR have not been described yet. It is expected that these results can serve as basis for raising discussions related to empiric treatment of bacterial vaginosis, considering limitations in the availability of diagnostic or antimicrobial therapy.

Submitted by Renata Lopes (renatasil82@gmail.com) on 2016-09-06T14:26:33Z No. of bitstreams: 1 arthurladeiramacedo.pdf: 2377553 bytes, checksum: 6c4d490fea818c6c85a6c551bd1e3d26 (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2016-09-06T14:50:02Z (GMT) No. of bitstreams: 1 arthurladeiramacedo.pdf: 2377553 bytes, checksum: 6c4d490fea818c6c85a6c551bd1e3d26 (MD5) / Made available in DSpace on 2016-09-06T14:50:02Z (GMT). No. of bitstreams: 1 arthurladeiramacedo.pdf: 2377553 bytes, checksum: 6c4d490fea818c6c85a6c551bd1e3d26 (MD5) Previous issue date: 2013-07-17 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Folhas de Gossypium barbadense L. (Malvaceae), conhecido como algodoeiro, são utilizadas popularmente para o tratamento de corrimento vaginal. A principal etiologia é a vaginose bacteriana (VB). VB é uma infecção complexa, na qual várias bactérias anaeróbias estão presentes, sendo a de maior incidência Gardnerella vaginalis. VB causa o aumento do corrimento vaginal, lise de células epiteliais vaginais, podendo causar parto prematuro (PPM), e outras complicações durante a gravidez. O mecanismo de PPM não está totalmente elucidado, porém uma proposta bem aceita é a de inflamação das membranas fetais. No presente trabalho avaliaram-se as atividades de extratos brutos e frações semi-purificadas de folhas de G. barbadense: 1) contra G. vaginalis, 2) citotóxica contra Artemia salina, 3) na viabilidade celular de células saudáveis (3T3) e cancerosas (MCF7) e 4) antiinflamatória no perfil de citocinas (IL-17, TNF- e INF-) e NO. Além disso, foi realizado o perfil cromatográfico dos extratos e frações semi-purificadas em CCD, CLAE-DAD e CG-EM e o isolamento de um flavonoide que foi identificado utilizandose RMN 1H, COSY e HSQC como sendo uma quercetina com substituição na hidroxila do carbono 3. Os extratos e frações polares mostram-se ricos em flavonoides, enquanto os apolares mostraram-se ricos em terpenoides e ácidos graxos saturados e insaturados. A fração diclorometânica (FDM) do extrato bruto metanólico (EBM) apresentou substâncias dos extratos polares e apolares. EBM e FDM apresentaram atividade expressiva contra G. vaginalis, aparentemente devido ao sinergismo entre flavonoides e substâncias apolares. EBM e FDM não apresentaram citotoxicidade contra A. salina e, em conjunto com as frações hexânica (FH), e em acetato de etila de EBM (FAE), apresentaram aumento da viabilidade de 3T3 e o extrato bruto aquoso e FDM reduziram a de MCF7. O extrato bruto hexânico (EBH) apresentou melhor atividade anti-inflamatória. Os demais extratos apresentaram menor atividade. Extratos e frações de folhas de G. barbadense, principalmente EBM e FDM, apresentam potencial para futuros estudos in vivo visando a confirmação das atividades em sistemas biológicos complexos. / Gossypium barbadense L. (Malvaceae) leaves, known as cotton, have been used for vaginal discharge treatment. Bacterial vaginosis (BV) is the major etiology. BV is a complex infection, involving a wide variety of anaerobes, being Gardnerella vaginailis the most incident. BV increases the amount of vaginal discharge, causing epithelial cells’ lysis, it can cause, also, preterm delivery (PTD) and other complications during pregnancy, as well. PTD mechanism is not totally known, but the inflammation of fetal membranes is one accepted theory. The present work evaluated 1) antimicrobial against G. vaginalis, 2) cytotoxicity against Artemia salina, 3) cell viability of health (3T3) and cancer (MCF7) cells and 4) anti-inflammatory evaluating cytokines (IL-17, TNF- e INF-) and NO activities of crude extracts and semi-purified fractions of G. barbadense leaves. Moreover, the chromatographic profile of crude extracts and semi-purified fractions were evaluated by TLC, HPLC-DAD and GC-MS. A flavonoid was isolated, being elucidated as a replaced quercetin, probably in hydroxyl group of carbon 3, by NMR 1H, COSY and HSQC. Flavonoids were abundant in polar extracts and fractions, and fatty acids and terpenoids were abundant in nonpolar ones. Polar and nonpolar compounds were observed in dichloromethane fraction (DMF) of methanol crude extract (MCE). MCE and DMF showed a expressive activity against G. vaginalis and the synergism between flavonoids and nonpolar substances appear to be crucial. MCE and DMF do not show cytotoxicity against A. salina, and with the hexane fraction (HF) and ethyl acetate fraction (EAF) of MCE showed an increase in cell viability of 3T3. Aqueous crude extract and DMF reduced the cell viability of MCF7. A most promising anti-inflammatory activity was presented by hexanic crude extract. Other crude extracts showed a less intense activity. G. barbadense leaves extracts and semi-purified fractions, chiefly MCE and DMF, presented good potential for future in vivo studies seeking the confirmation of these activities in complex biological systems.

<p>Risk factors such as intrauterine and vaginal infection put pregnant women at risk for delivering preterm. Bacterial vaginosis (BV) is a polymicrobial clinical syndrome commonly diagnosed in women of reproductive age, with women of African descent with low socioeconomic status and previous preterm delivery at high risk. Although frequently isolated from healthy women,&nbsp / Gardnerella vaginalis has been most frequently associated with BV. There is limited data available on the prevalence of BV in Southern Africa / therefore, we embarked on a study to determine the&nbsp / prevalence of BV and G. vaginalis in predominantly black communities in the Western Cape, in order to establish the role of G. vaginalis in BV. Women attending various Maternity and Obstetrics&nbsp / units (MOU) in the Cape Peninsula with and without a history of pre-term delivery (PTD) were invited to participate in the study. Several factors were statistically associated with pregnancy history,&nbsp / including location of study population, parity, smoking and presence of clinical symptoms. The presence of G. vaginalis was determined by culture in 51.7% of the preterm delivery group (PTDG)&nbsp / and 44% of the full-term delivery group (FTDG) women. BV was detected in 31.13% of PTDG and 23.67% of FTDG by Gram stained analysis according to Nugent scoring criteria, with age and HIV&nbsp / status posing as risk factors. When comparing PTDG and FTDG for an association between the presence of G. vaginalis and BV, a stronger association was observed in the PTDG but it was not statistically significant. In both PTDG and FTDG, G. vaginalis was isolated significantly more often in women diagnosed with BV at 24.5% (p &lt / 0.05). Antibiogram studies revealed both Metronidazole and Clindamycin resistant strains of G. vaginalis. G. vaginalis Biotype 7 is specifically associated with BV, while Biotype 2 appears to be associated with BV in women with a history&nbsp / of PTD. Accuracy of diagnostic tools were tested and it was determined that Nugent scoring is more sensitive in diagnosing BV (76.04%), but culture for G. vaginalis is more specific (83.21%). Although this study was limited in that we were unable to follow-up pregnancy outcomes, we were able to confirm the perceived role of G. vaginalis in BV.&nbsp / </p>

<p>Risk factors such as intrauterine and vaginal infection put pregnant women at risk for delivering preterm. Bacterial vaginosis (BV) is a polymicrobial clinical syndrome commonly diagnosed in women of reproductive age, with women of African descent with low socioeconomic status and previous preterm delivery at high risk. Although frequently isolated from healthy women,&nbsp / Gardnerella vaginalis has been most frequently associated with BV. There is limited data available on the prevalence of BV in Southern Africa / therefore, we embarked on a study to determine the&nbsp / prevalence of BV and G. vaginalis in predominantly black communities in the Western Cape, in order to establish the role of G. vaginalis in BV. Women attending various Maternity and Obstetrics&nbsp / units (MOU) in the Cape Peninsula with and without a history of pre-term delivery (PTD) were invited to participate in the study. Several factors were statistically associated with pregnancy history,&nbsp / including location of study population, parity, smoking and presence of clinical symptoms. The presence of G. vaginalis was determined by culture in 51.7% of the preterm delivery group (PTDG)&nbsp / and 44% of the full-term delivery group (FTDG) women. BV was detected in 31.13% of PTDG and 23.67% of FTDG by Gram stained analysis according to Nugent scoring criteria, with age and HIV&nbsp / status posing as risk factors. When comparing PTDG and FTDG for an association between the presence of G. vaginalis and BV, a stronger association was observed in the PTDG but it was not statistically significant. In both PTDG and FTDG, G. vaginalis was isolated significantly more often in women diagnosed with BV at 24.5% (p &lt / 0.05). Antibiogram studies revealed both Metronidazole and Clindamycin resistant strains of G. vaginalis. G. vaginalis Biotype 7 is specifically associated with BV, while Biotype 2 appears to be associated with BV in women with a history&nbsp / of PTD. Accuracy of diagnostic tools were tested and it was determined that Nugent scoring is more sensitive in diagnosing BV (76.04%), but culture for G. vaginalis is more specific (83.21%). Although this study was limited in that we were unable to follow-up pregnancy outcomes, we were able to confirm the perceived role of G. vaginalis in BV.&nbsp / </p>

The vaginal microbial community is critical to a woman’s health and the health of her family. Bacterial vaginosis (BV) is a polymicrobial syndrome characterized by a shift of the vaginal microbiota from a Lactobacillus dominated community to a dense biofilm containing a complex mixture of organisms. Although BV is an important risk factor for poor reproductive health outcomes, the etiology of BV is poorly understood. Gardnerella vaginalis is a hallmark species of BV. Phylogenetic analysis of cpn60 universal target sequences from metagenomic studies of the vaginal microbiome and from G. vaginalis isolates resolved four subgroups within the species. This subdivision, supported by whole genome similarity comparisons, demonstrated that these subgroups might represent different species. Among a group of African women, only G. vaginalis subgroup B was significantly more abundant in women with BV relative to women with Nugent scores not consistent with BV. To characterize the subgroups further, several phenotypic and molecular factors of G. vaginalis subgroups were assessed. Proteomic profiles of isolates within each subgroup formed unambiguous clusters. Sialidase gene sequences were detected in all subgroups, however enzymatic activity was detected only in subgroup B. Two isolates of subgroup B isolates (N153 and N101) were incapable of growth in 7% CO2. Given the well-known relationship between an anaerobic microbiota and BV, anaerobic isolates of G. vaginalis are potentially important players in the vaginal microbial community. To determine genome content differences that could account for the phenotypic difference, whole genome sequences of four G. vaginalis subgroup B isolates representing facultative and anaerobic phenotypes were determined. Comparison of genomes led to the identification of genes predicted to encode proteins involved in cell wall biogenesis and protection from oxidative damage that might account for the observed phenotypes. The cpn60 universal target based methodology that improved resolution of the vaginal microbiota including G. vaginalis was applied in a prospective study of the vaginal microbiome of women with preterm premature rupture of membranes (PPROM). The objectives were to characterize the vaginal microbiota of women following PPROM, and to determine if microbiome composition at the time of rupture predicts latency duration and perinatal outcomes. Only 13/70 samples collected from 36 women were dominated by Lactobacillus spp., the expected profile for healthy women, while Megasphaera type 1 and Prevotella spp. were detected in all samples. Microbiome profiles at the time of membrane rupture did not cluster by gestational age at PPROM, or latency duration. Microbial profiles were unstable over the latency period, with dramatic shifts in composition between weekly samples, and an overall decrease in Lactobacillus abundance. Mollicutes were detected by PCR in 81% (29/36) of women, and these women had significantly lower gestational age at delivery and correspondingly lower birth weight infants than Mollicutes negative women. Taken together, the results presented in this thesis demonstrate the value of high resolution profiling of the vaginal microbiome using cpn60 UT sequences. The resolution of subgroups within G. vaginalis has potentially significant implications for women's health diagnostics, requiring a shift away from considering G. vaginalis as a single entity. The PPROM study provides foundational information that may lead to the identification of informative sequence patterns, providing clinicians with better tools for expectant management following PPROM.