Philosophiae Doctor - PhD / The aim of the study was to detect the prevalence of AV and its associated bacteria with preterm delivery in the Western Cape, South Africa. Furthermore, it sought particularly to examine and investigate the predictive value of GBS and E. faecalis for preterm delivery (PTD). It also aimed to establish other factors which may predict adverse pregnancy outcomes. Three hundred and one pregnant women were recruited from four different antenatal in the Western Cape, South Africa. The study conformed with the Declaration of Helsinki (2013). Maternal data was collected from a questionnaire and maternal medical records. Vaginal and rectal swabs were collected and microscopically examined for AV, followed by culture characterization of GBS and E. faecalis. Antimicrobial susceptibility testing was also performed. In this study, AV was detected in 79 (26.2%) of the 301 pregnant women, and GBS and E. faecalis isolated from 50 (16.6%) and 21 (7.0%) respectively. GBS serotype V was the predominant serotype, followed by serotype III. Pulse field gel electrophoresis (PFGE) profile analysis for both GBS and E. faecalis yielded a total of 24 restrictions profiles for GBS and 16 for E. faecalis. Multivariable analysis revealed that parity, gravidity, vaginal discharge, urinary tract infection, and smoking were significantly associated with PTD. The results from the study provides improved guidelines maternal screening of pregnant women. The early detection of AV-related bacteria may significantly reduce maternal and neonatal morbidity.

Source: Masters Abstracts International, Volume: 47-06, page: 3500. Adviser: Kim Waller. Includes bibliographical references.

La vaginosis bacteriana (VB) es actualmente una de las causas más comunes de infección vaginal en mujeres en edad fértil, sin embargo, poco es lo que se sabe sobre su historia natural y las causas que la generan. En este sentido, se realizó un estudio analítico descriptivo, retrospectivo del tipo casocontrol para determinar los factores de riesgo asociados a la vaginosis bacteriana (VB) en pacientes atendidas en la clínica Good Hope. Utilizando la puntuación de Nugent se analizaron las muestras de secreción vaginal de las pacientes atendidas en la clínica durante el periodo julio–octubre de 2017. De las 673 muestras analizadas, el 61,1% de pacientes presentaban una microbiota normal y el 18,4% presentaban VB, a partir de estos dos grupos se seleccionaron aleatoriamente 60 pacientes con VB para conformar el grupo de los casos y 120 pacientes sin VB para el grupo de los controles. A partir de la historia clínica se colectaron en una ficha los datos clínicos y demográficos de cada paciente. Estos datos fueron procesados y por medio del análisis bivariado y multivariado, donde se evaluó el riesgo y grado de asociación con la VB. No se encontró diferencias significativas en cuanto a la edad en ambos grupos; en cuanto a la residencia, los distritos de Santiago de Surco y Cercado de Lima fueron los más prevalentes para el grupo de los casos y de los controles respectivamente. Tras el análisis estadístico los factores: estado de gestación (OR= 0,483, IC= 0,244–0,955, p=0,036) y el antecedente de Candidiasis (OR= 3,651 IC= 1,055–12,630, p=0,041) tuvieron significancia estadística como factor protector y de riesgo para la VB, respectivamente. / Tesis

Estrus synchronization (ES) is a valuable technology that can help beef cow-calf producers to overcome infertility caused by prolonged anestrus. Protocols for ES that use progesterone (P4) supplementation are of particular value to cows with prolonged postpartum anestrus as P4 triggers them to begin cycling and allows them to have fertility similar to that of cycling cows. Supplementation of P4 intravaginally with the use of a controlled internal drug release device (CIDR) improves cycle induction when compared to oral administration of P4. Vaginitis has been reported as a side effect to CIDR use in cattle, which raises concerns about its downstream effects on fertility. More specifically, the effects of CIDR use on the vaginal environment requires exploration, as no studies have explored the changes in vaginal microbiome in response to CIDR based ES protocols. In cattle, the vaginal microbiome has not been widely explored. On the contrary, the human vaginal microbiome is a well-defined environment, rich in bacteria from the genus Lactobacillus, which are responsible for promoting an environment of acidic pH. The dominance of Lactobacillus in the human vagina, however, fluctuates according to steroid hormone concentrations, and disruptions in the vaginal environment will cause depletion of Lactobacillus species, increase in vaginal pH and decreased fertility. Based on this data in humans, our objectives were to describe incidence of vaginitis caused by the CIDR in beef cows, as well as the vaginal microbiome changes in response to CIDR based protocols, and explore their relation to fertility. We found high incidences of vaginitis caused by CIDR use, yet CIDR-induced vaginitis did not negatively affect pregnancy outcomes. However, at CIDR withdrawal, there was decreased bacterial diversity, increased vaginal pH, increased bacterial abundance, and increased vaginal inflammation when compared to what was observed prior to CIDR insertion. Furthermore, abundance of bacteria, vaginal inflammation, and bacterial diversity, but not vaginal pH, were restored to normal values by the day of timed artificial insemination. This important finding suggests that although the vaginal microbiome was disrupted by the use of CIDR, the vaginal microbiome is resilient and capable of restoring its natural conditions without intervention. Finally, cows that ultimately became pregnant were found to have had increased bacterial diversity and decreased vaginal pH prior to protocol initiation, suggesting that the vaginal microbiome may play a role in individual cow fertility. In conclusion, our results indicate that for beef cows a more diverse vaginal microbiome with decreased vaginal pH presents greater resilience of the microbiome towards disruptions caused by an ES protocol, which is translated in greater pregnancy success. / Doctor of Philosophy / According to the Food and Agriculture Organization, the world population is expected to grow by 51% by the year of 2100. The efficiency of food production must therefore be optimized, given the finite availability of farmable land. In beef production, cow fertility is of great importance, since it will ultimately determine the number of animals available for slaughter. The main reproductive issue that cow-calf producers face is that after calving, cows will undergo a period known as postpartum anestrus, in which cows fail to ovulate. Artificial insemination (AI) can help to optimize beef production efficiency, since it allows for the dispersal of semen from valuable bulls to facilitate the genetic enhancement of herds. The use of estrus synchronization (ES) protocols allows for induction and synchronization of ovulation, which allows AI to be performed at the same time for large groups of cows. Progesterone is often used in ES protocols and is the hormone responsible for inducing cyclicity in postpartum cows. Progesterone can be administered either orally or intravaginally via the use of a controlled internal drug release (CIDR). While the CIDR seems to be more effective at inducing cyclicity of cattle compared to oral progesterone administration, vaginal inflammation as response to the CIDR has been reported in cattle. Little is known about the downstream effects of this inflammation on the normal vaginal microbiota and fertility in cattle. In humans, the vaginal microbiome is predominated by a single genus of bacteria (Lactobacillus), that has an essential role in producing lactic acid, which results in the human vagina being remarkably acidic. In humans, depletion of this bacteria, a condition called bacterial vaginosis (BV), allows for other types of bacteria to grow, which results in an increased vaginal pH and decreased fertility. The bovine vaginal microbiome composition and pH in response to the hormones administered during ES protocols and its relation to fertility have not been widely explored. Our objectives were to document the incidence of vaginitis caused by the CIDR in beef cows and evaluate its effects on the vaginal microbiome changes and fertility. We found high incidences of vaginitis caused by the CIDR, yet no effects of CIDR-induced vaginitis were seen on pregnancy success to the protocol. However, decreased bacterial diversity, followed by increases in vaginal pH, abundance of bacteria and vaginal inflammation are all detected at CIDR withdrawal when compared to before CIDR insertion. Furthermore, abundance of bacteria, vaginal inflammation, and bacterial diversity, but not vaginal pH, were restored to normal values by the day of timed AI, indicating that although the vaginal microbiome was disrupted using CIDR, the vaginal microbiome can restore to natural conditions, and indicate resilience of the vaginal microbiome. Finally, cows that became pregnant to the protocol presented increased bacterial diversity and decreased vaginal pH prior to the protocol. In conclusion, our results indicate that for beef cows a more diverse vaginal microbiome with decreased vaginal pH presents greater resilience of the microbiome towards disruptions caused by an ES protocol, which is translated in greater pregnancy success.

Vaginitis is a vaginal infection which affects many women all over the world. The disorder is characterized by an infection of the vaginal area which can cause problems like abnormal vaginal discharge, itching and redness. The two most common causes of vaginitis are bacterial vaginosis and Candida vaginitis. The prevalence of bacterial vaginosis in Sweden is around 10-20 % and approximately 75 % of all women will once in their lifetime suffer from vaginal yeast infection. The clinical symptoms of vaginal infections are not specific and the diagnosis methods of bacterial vaginosis and Candida vaginitis are subjective and depended on the acuity of the clinician. Due to the lack of standardized and objective diagnostic tools, misdiagnosis and consequently incorrect treatment may occur. As vaginal infections and symptoms impact greatly of women´s quality of life and vaginitis have been associated with serious public health consequences, it is essential to diagnose and treat the conditions correctly. Hence, there is a great need of better methods of diagnosing these conditions. The aim of this master thesis was to develop quantitative species-specific real-time PCR assays to use in diagnosing the two most common causes of vaginitis i.e. bacterial vaginosis and Candida vaginitis. Potential markers for bacterial vaginosis (Atopobium vaginae, BVAB2, Gardnerella vaginalis, Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus jensenii, Lactobacillus iners, Megasphaera type 1, Megasphaera type 2, Mobiluncus curtisii, Mobiluncus mulieris and Leptotrichia/Sneathia species) and Candida vaginitis (Candida albicans, Candida glabrata, Candida parapsilosis and Candida tropicalis) were chosen. Primers and probes were designed and tested on reference strains and vaginal samples. Single- and multiplex PCR reactions were successfully optimized with the designed oligonucleotides. Furthermore, standard curves with excellent linearity were created and covered more than five orders of magnitude. These developed quantitative species-specific real-time PCR assays will, in a prospective medical validation, quantify 300 vaginal samples from women visiting the RFSU Clinic in Stockholm.

A human safe, Alhydrogel adjuvanted whole cell Trichomonas vaginalis vaccine was tested for efficacy in a BALB/c mouse model of vaginal infection. Additionally, the systemic and local immune response were measured. Vaccination reduced incidence and increased clearance of infection, and induced both systemic and local humoral immune responses. CD4+ cells were detected in vaginal tissues following intravaginal challenge with T. vaginalis, but were not seen in uninfected mice. CD4+ cells were detected more often, earlier, and in greater numbers in vaccinated vaginal tissues compared to unvaccinated controls. Presence of CD4+ T cells following infection can have significant implications of increasing HIV susceptibility and transmission. These data suggest that the vaccine induces local and systemic immune responses, and confers significantly greater protection against vaginal challenge than unvaccinated vaginal challenge. These data support the potential for a human vaccine against T. vaginalis infection that could also impact the incidence of HIV infections.

A human safe, Alhydrogel adjuvanted whole cell Trichomonas vaginalis vaccine was tested for efficacy in a BALB/c mouse model of vaginal infection. Additionally, the systemic and local immune response were measured. Vaccination reduced incidence and increased clearance of infection, and induced both systemic and local humoral immune responses. CD4+ cells were detected in vaginal tissues following intravaginal challenge with T. vaginalis, but were not seen in uninfected mice. CD4+ cells were detected more often, earlier, and in greater numbers in vaccinated vaginal tissues compared to unvaccinated controls. Presence of CD4+ T cells following infection can have significant implications of increasing HIV susceptibility and transmission. These data suggest that the vaccine induces local and systemic immune responses, and confers significantly greater protection against vaginal challenge than unvaccinated vaginal challenge. These data support the potential for a human vaccine against T. vaginalis infection that could also impact the incidence of HIV infections.

Objetivo: Avaliar a citologia cérvico-vaginal em adolescentes com lúpus eritematoso sistêmico juvenil (LESJ) e comparar com controles. Material e métodos: Cinqüenta e duas adolescentes com LESJ (critérios do American College of Rheumatology) foram comparadas com 52 controles saudáveis. Todos os esfregaços de Papanicolaou foram avaliados por uma mesma citopatologista, que desconhecia o exame ginecológico, e foram classificados de acordo com o Sistema de Bethesda, 2001. Resultados: As médias das idades das pacientes com LESJ e controles foram similares (16,17 ± 1,94 versus 16,13 ± 2,16 anos, p=0,92). A citologia cérvico-vaginal foi similar em ambos os grupos, embora as relações sexuais no último mês tenham sido menos freqüentes nas pacientes com LESJ em relação aos controles (23% versus 59,6%, p=0,0003). Apenas uma paciente (2%) com LESJ e duas controles (4%) tinham displasia cervical (LIE-BG) e papilomavírus humano (HPV) (p=1,0). Citologia cérvico-vaginal inflamatória foi observada em 21 (60%) das pacientes com SLEDAI maior ou igual a 4 e em apenas 4 (23%) daqueles com SLEDAI < 4 (p=0,001). Assim como, uma maior freqüência de achados inflamatórios também foi observada em adolescentes virgens com LESJ (57% versus 8%, p=0,005). Vaginite por Candida spp foi observada em 7 pacientes com LESJ (14%) e em nenhuma dos controles (p=0,012), e foi associada com uso de drogas imunossupressoras (p=0,01) e dose alta de prednisona (p=0,002). Conclusão: Nossos achados indicam que o trato genital feminino é um órgão alvo no LESJ, pois inflamação cérvico-vaginal está associada com atividade da doença independentemente da atividade sexual. / Objective: To evaluate cervicovaginal cytology in adolescents with juvenile systemic lupus erythematosus (JSLE) and to compare them to controls. Material and methods: Fifty-two female adolescents with JSLE (American College of Rheumatology criteria) were compared to 52 age-matched healthy controls. All Pap smears were evaluated by the same cytopathologist blinded to gynecology examination, and performed according to the Bethesda Classification System 2001. Results: The mean age of JSLE patients and controls were similar (16.17 ± 1.94 vs. 16.13 ± 2.16 years, p=0.92). The cervicovaginal cytology was found to be similar in both groups, although sexual intercourses in the last month were less frequent in JSLE than controls (23% vs. 59.6%, p=0.0003). Only one patient (2%) with JSLE versus two controls (4%) had cervical dysplasia (LGSIL) and human papilomavirus (p=1.0). Inflammatory cervicovaginal cytology was observed in 21 (60%) of patients with SLEDAI ? 4 and only 4(23%) of those with SLEDAI<4 (p=0.001). Likewise, a higher frequency of inflammatory changes were also observed in virgin JSLE (57% vs. 8%, p=0.005). Candida spp vaginitis was observed in 7 JSLE (14%) versus none in controls (p=0.012) and was associated to immunosuppressive drugs (p=0.01) and high dose of prednisone (p=0.002). Conclusion: Our findings supports the notion that female genital tract is a target organ in SLE since cervical inflammation is associated to disease activity independently of sexual activity.

Objetivo: Avaliar a citologia cérvico-vaginal em adolescentes com lúpus eritematoso sistêmico juvenil (LESJ) e comparar com controles. Material e métodos: Cinqüenta e duas adolescentes com LESJ (critérios do American College of Rheumatology) foram comparadas com 52 controles saudáveis. Todos os esfregaços de Papanicolaou foram avaliados por uma mesma citopatologista, que desconhecia o exame ginecológico, e foram classificados de acordo com o Sistema de Bethesda, 2001. Resultados: As médias das idades das pacientes com LESJ e controles foram similares (16,17 ± 1,94 versus 16,13 ± 2,16 anos, p=0,92). A citologia cérvico-vaginal foi similar em ambos os grupos, embora as relações sexuais no último mês tenham sido menos freqüentes nas pacientes com LESJ em relação aos controles (23% versus 59,6%, p=0,0003). Apenas uma paciente (2%) com LESJ e duas controles (4%) tinham displasia cervical (LIE-BG) e papilomavírus humano (HPV) (p=1,0). Citologia cérvico-vaginal inflamatória foi observada em 21 (60%) das pacientes com SLEDAI maior ou igual a 4 e em apenas 4 (23%) daqueles com SLEDAI < 4 (p=0,001). Assim como, uma maior freqüência de achados inflamatórios também foi observada em adolescentes virgens com LESJ (57% versus 8%, p=0,005). Vaginite por Candida spp foi observada em 7 pacientes com LESJ (14%) e em nenhuma dos controles (p=0,012), e foi associada com uso de drogas imunossupressoras (p=0,01) e dose alta de prednisona (p=0,002). Conclusão: Nossos achados indicam que o trato genital feminino é um órgão alvo no LESJ, pois inflamação cérvico-vaginal está associada com atividade da doença independentemente da atividade sexual. / Objective: To evaluate cervicovaginal cytology in adolescents with juvenile systemic lupus erythematosus (JSLE) and to compare them to controls. Material and methods: Fifty-two female adolescents with JSLE (American College of Rheumatology criteria) were compared to 52 age-matched healthy controls. All Pap smears were evaluated by the same cytopathologist blinded to gynecology examination, and performed according to the Bethesda Classification System 2001. Results: The mean age of JSLE patients and controls were similar (16.17 ± 1.94 vs. 16.13 ± 2.16 years, p=0.92). The cervicovaginal cytology was found to be similar in both groups, although sexual intercourses in the last month were less frequent in JSLE than controls (23% vs. 59.6%, p=0.0003). Only one patient (2%) with JSLE versus two controls (4%) had cervical dysplasia (LGSIL) and human papilomavirus (p=1.0). Inflammatory cervicovaginal cytology was observed in 21 (60%) of patients with SLEDAI ? 4 and only 4(23%) of those with SLEDAI<4 (p=0.001). Likewise, a higher frequency of inflammatory changes were also observed in virgin JSLE (57% vs. 8%, p=0.005). Candida spp vaginitis was observed in 7 JSLE (14%) versus none in controls (p=0.012) and was associated to immunosuppressive drugs (p=0.01) and high dose of prednisone (p=0.002). Conclusion: Our findings supports the notion that female genital tract is a target organ in SLE since cervical inflammation is associated to disease activity independently of sexual activity.

Made available in DSpace on 2015-07-22T20:49:11Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-06-30 / Introdução - Os Streptococcus agalactiae são bactérias consideradas parte da microbiota da vagina, das vias aéreas superiores e do trato intestinal, porém sua importância como patógeno humano vem crescendo no decorrer dos anos. Quando presente na vagina podem trazer sérias consequências para a mulher, podendo estar associados à infertilidade, aumentar o risco de ruptura prévia de membrana, levar a corioaminionite, endometrite, etc. e, a partir da colonização vaginal da mãe, pode também haver contaminação dos neonatos na hora do parto. Objetivo - O objetivo deste estudo foi determinar a associação dos Streptococcus agalactiae com inflamação vaginal em mulheres atendidas nos laboratórios públicos de Maceió. Material e método - Foi coletado material do terço médio distal da vagina de 1014 mulheres entre 18 e 49 anos. Um dos swabs foi utilizado para a realização de exame bacterioscópico e os outros dois swabs foram utilizados para a realização de cultura para o Streptococcus agalactiae (colocado em caldo Todd-Hewitt) e outros microrganismos, além de exame a fresco direto. A identificação dos microrganismos foi realizada de acordo com a literatura. Resultados - A frequência de colonização vaginal pelo Streptococcus agalactiae encontrada neste estudo foi de 13,9%, sendo que 98 (9,7%) culturas foram positivas somente para este microrganismo e em 43 (4,2%) o mesmo estava associado a outros microrganismos. Dentre as mulheres com cultura positiva apenas para o estreptococo, 18,9% apresentaram reação inflamatória, enquanto somente 6,7% não tinham inflamação. A proporção de reações inflamatórias observadas nas mulheres colonizadas pelo estreptococo foi comparável à de outros patógenos comumente encontrados na vagina como: Candida spp., G. vaginalis e Trichomonas spp.. Em relação ao grau de associação destes microrganismos com inflamação vaginal, verificou-se um Risco Relativo de 2,80 para o Streptococcus agalactiae, 2,01 para Candida spp., 0,55 para Gardnerella vaginalis e 19,03 para Trichomonas spp. Estes dados foram confirmados pela análise multivariada pelo método Forward Stepwise (Wald). Foi verificado que a gestação não constitui um fator de risco, bem como a idade, número de parceiros sexuais, frequência de relações sexuais, abortos e gestações anteriores. Conclusão - O estreptococo do grupo B, quando presente na vagina, mostrou ser um agente promotor de inflamação, a exemplo de outros microrganimos como Candida spp e Trichomonas spp, que são sempre valorizados quando encontrados nesse sítio. Este fato mostra que esta bactéria, ainda considerada parte da microbiota vaginal, pode ser considerada tão patogênica como outros microrganismos, havendo assim a necessidade de diferenciar quanto a um estado de simples colonização ou se a bactéria está sendo agente de processo infeccioso. / superior aerial tract, and intestinal tract microbiota, but it is growing in importance as a human pathogen throughout the last years. When it is present in vagina, it can bring serious consequences to the woman, and can be associated to infertility, improving risk of premature rupture of membrane, leading to chorioamnionitis, endometritis, etc. and from mother vaginal colonization there may be contamination of neonates during delivery. Objective - Objective of this study was to determine the association of Streptococcus agalactiae with vaginal inflammatory process in women attended in public laboratories of Maceió. Material and methods – It has been collected material in the distal medium third of the vagina of 1014 women from 18 to 49 years. One of the swabs has been used to bacterioscopic examination procedures and the other two swabs had been used for culture procedures of Streptococcus agalactiae (put in Todd-Hewitt broth) and other microorganisms, beyond the fresh direct examination. The identification of the microorganisms has been made according to literature. Results – The frequency of vaginal colonization by Streptococcus agalactiae found in this study has been of 13,9%, being 98 (9,7%) positive cultures only for this microorganism and 43 (4,2%) associated with other microorganisms. Among women with positive culture only to streptococcus, 18,9% presented inflammatory reaction, while only 6,7% didn’t have inflammation. The proportion of inflammatory reactions observed in women colonized by the streptococcus has been comparable to that of other pathogens commonly found in vagina as: Candida spp., G. vaginalis and Trichomonas spp.. In relation to the degree of association of those microorganisms with vaginal inflammatory processes, it has been verified a Relative Risk of 2,80 for Streptococcus agalactiae, 2,01 for Candida spp., 0,55 for Gardnerella vaginalis and 19,03 for Trichomonas spp. Those data have been confirmed by the multivariant analysis using Forward Stepwise method (Wald). It has been verified that pregnancy does not constitute a risk factor, nor age, number of partners, frequency of intercourse, abortions and previous pregnancies. Conclusion – B group streptococcus, when it is present in vagina showed to be an inflammatory process promoting agent, as for example other microorganisms, like Candida spp and Trichomonas spp, that are always valued when they are found in that site. This fact shows that this bacteria that is still considered as part of vaginal biota, should be considered so much pathogenic as other microorganisms, thus existing the necessity of differenciating between a state of simple colonization or otherwise if the bacteria is being an infectious process agent. / TEDE / BV UNIFESP: Teses e dissertações