The human vaginal ecosystem is dominated by Lactobacillus species. An altered vaginal flora can result in symptomatic conditions such as bacterial vaginosis and vulvo-vaginal candidiasis, and urogenital colonisation by uropathogenic bacteria can cause urinary tract infection. The protective role of lactobacilli is gradually being accepted and clinical studies have been carried out in order to evaluate the use of promising probiotic bacteria, which are defined as “live microorganisms which when administered in adequate amounts confer a health benefit on the host”. This thesis includes an investigation into the ecological role of lactobacilli in the genital tract in healthy women, with respect to the relationship to other species and vaginal pH. Furthermore, in order to find different probiotic strains with promising probiotic qualities, Lactobacillus strains were screened in two diverse screening processes. The selected strains were further evaluated in clinical trials. The prevalence of group B streptococci (GBS) and yeast was significantly dependent on the number of vaginal lactobacilli among healthy women. GBS were less frequently found in women with high numbers of vaginal lactobacilli than in women with low numbers and the prevalence of yeast was significantly higher in women with 3-6.99 log10 lactobacilli sample-1 than in women with less than 3 or ≥7 log10 lactobacilli sample-1. Furthermore, the first screening made on 511 strains isolated from the female genital tract resulted in the final selection of a Lactobacillus plantarum, designated LB931. The screening showed that LB931 had a strong technical growth, survived through freeze-thawing, produced substances bactericidal to uropathogenic bacteria and was a rapid and strong producer of hydrogen peroxide. Further characterisation showed that LB931 possessed the properties required for probiotics with the capability to prevent urogenital infections. LB931 could be supplied to the genital tract through the usage of panty liners impregnated with the strain. In the second screening, Lactobacillus fermentum, designated Ess-1, was the only one out of 126 Lactobacillus strains with strong capacity to inhibit Candida albicans and Candida glabrata. Additional characterisation showed that L. fermentum Ess-1 had the properties that are needed to prevent over-growth of Candida in the vulvo-vaginal tract. The result of the case study showed that a high and frequent dosage of Ess-1 is needed and that improved vulvo-vaginal candidiasis specific diagnostic criteria are required. In conclusion, L. plantarum LB931 and L. fermentum Ess-1 are promising probiotic strains to be used in the prevention of recurrent urogenital infections in women and to enhance the normal flora in healthy women.

Submitted by Fabio Sobreira Campos da Costa (fabio.sobreira@ufpe.br) on 2016-05-09T14:32:17Z No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Carmen Marangoni_PPGBV_CCB_2015.pdf: 2902792 bytes, checksum: be5b9acdf98bf4f4ba08a353a0ed4480 (MD5) / Made available in DSpace on 2016-05-09T14:32:17Z (GMT). No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Carmen Marangoni_PPGBV_CCB_2015.pdf: 2902792 bytes, checksum: be5b9acdf98bf4f4ba08a353a0ed4480 (MD5) Previous issue date: 2015-04-15 / CNPq / As infecções das vias gênito-urinárias são uma realidade muito difundida mundialmente. São causadas por bactérias, fungos e protozoários que colonizam os tecidos urogenitais. Existem muitos produtos naturais à base de plantas que demostraram ter eficácia no tratamento desses distúrbios. No Brasil, em muitas regiões, o emprego das plantas medicinais é prática comum. Estudos etnobotânicos realizados no Nordeste do Brasil com populações tradicionais, incluindo tribos indígenas, indicam que muitas plantas são utilizadas para tratar distúrbios das vias gênito-urinárias. Para melhor entender as práticas de cura tradicionais e corroborar as suas eficácia, neste estudo avaliou-se o potencial antimicrobiano in vitro de uma seleção de plantas medicinais tradicionalmente utilizadas para tratar esses tipo de transtorno por duas populações indígenas, os Pankararu e os Fulni-ô, que habitam a região semiárida do Nordeste do Brasil. As plantas foram coletadas no final da estação chuvosa na comunidade rural de Riachão de Malhada de Pedra, município de Caruaru. Extratos aquosos e hidroalcoólicos foram preparados com a parte da planta tradicionalmente utilizada e a avaliação da atividade antimicrobiana foi realizada com a técnica da microdiluição, testando os extratos frente microrganismos capazes de colonizar o aparelho urogenital (Candida albicans, C. tropicalis, Enterococcus faecalis, Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa, Staphylococcus aureus, S. saprophyticus). A avaliação da atividade anti- Trichomonas vaginalis foi realizada através do teste da viabilidade dos trofozooides. Foi realizado o perfil fitoquímico e avaliada a atividade antioxidante dos extratos obtidos da forma tradicional. Foi também testada a correlação entre atividade antimicrobiana e uso tradicional, reportado como índice de Importância Relativa de Uso. Os resultados mostraram que os microrganismos mais susceptíveis foram os dois pertencentes ao gênero Staphylococcus, sendo os extratos aquoso e hidroalcoólico de Maytenus rigida e Spondias tuberosa os mais ativos (MIC = 0.2 mg/mL). As plantas cujos extratos da casca demonstraram maior espectro de ação foram Anacardium occidentale, Myracrodruon urundeuva e S. tuberosa. A maioria dos extratos testados mostrou atividade frente ao protozoário T. vaginalis. Os extratos de Sideroxylon obtusifolium mostraram uma toxicidade contra ao parasita comparável à do metrodinazol. O teor de taninos encontrado nos extratos mostrou ter uma correlação significativa com a atividade antimicrobiana, indicando um papel dessa classe de compostos na atividade aqui registrada. Encontrou-se também uma correlação significativa entre a atividade antimicrobiana e o uso tradicional. As espécies que apresentam maior atividade antioxidante foram Anadenanthera colubrina, M. urundeuva, S. tuberosa que coincidiram com as espécies que apresentaram os maiores teor de fenóis totais. O teor de flavonoides não parece estar correlato com a atividade antioxidante. Os resultados aqui apresentados apontam que as espécies A. occidentale, M. urundeuva e S. tuberosa produzem substâncias ativas frente a todos os microrganismos testados, justificando o uso tradicional. Essas três espécies, juntas com M. rigida e S. obtusifolium mostraram atividades promissora para o desenvolvimento de novos tratamentos contra T. vaginalis. / The urogenital infections are a common reality worldwide, caused by bacteria, yeasts and protozoans which are able to colonize the urogenital tissues. Several plant-based products demonstrated efficacy in treating this kind of disturbs. Ethnopharmacological surveys realized with traditional populations, including indigenous ones, in the Northeastern semiarid region of Brazil, showed that several plant species are utilized to treat urinary and genital tracts infections. To better understand the traditional medicine practices and corroborate their efficacy, in this study the antimicrobial potential of eight medicinal plants used to treat urogenital infections by two indigenous tribes, Pankararu e Fulni-ô, located in the Northeastern Brazil, was tested in vitro. The plant material was collected at the end of the rain season in the rural community Riachão de Malhada de Pedra, Caruaru district, in the agreste of the Pernambuco state. Aqueous and hidroalcoholic extracts were obtained using the part of the plant traditionally used. The antimicrobial assay was performed with the microdiluition method against human pathogens able to colonize the urogenital system (Candida albicans, C. tropicalis, Enterococcus faecalis, Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa, Staphylococcus aureus, S. saprophyticus). The anti-Trichomonas vaginalis activity was evaluate testing the viability of the trophozoites. The phytochemical profile of the extracts was realized and the antioxidant activity was measured too. Furthermore the correlation between the antimicrobial activity and the traditional use, using the Relative Use Importance index, was tested. The results showed that the most susceptive of the tested microorganism were the two Staphylococcus species, being the aqueous and hidoalcoholic extracts of Maytenus rigida and Spondias tuberosa the most active ones (MIC = 0,2 mg/mL). Anacardium occidentale, Myracrodruon urundeuva and S. tuberosa bark extracts were found to have the broadest spectrum of activity. The majority of the extracts showed an anti-T. vaginalis activity. Sideroxylon obtusifolium extracts showed the higher activity toward the parasite, comparable with metrodinazol one. A positive correlation was encountered between the quantity of tannins and the antimicrobial activity, this indicates the role of this class of compounds in the activity here reported. A significant correlation was also found between the antimicrobial activity and the traditional use. The species presenting the higher antioxidant activities were Anadenanthera colubrina, M. urundeuva, S. tuberosa, which are the species with the higher total phenols content. No correlation between flavonoids content and antioxidant activity was found. The results of this study indicate that A. occidentale, M. urundeuva e S. tuberosa synthetize substances which act towards all the tested microorganism, justifing the traditional use of these medicinal plants. These three species, along with M. rigida and S. tuberosa showed a strong anti-T. vaginalis activity, which is promising for the development of new treatments for this protozoan infections.