Submitted by Renata Lopes (renatasil82@gmail.com) on 2015-12-01T10:18:13Z No. of bitstreams: 1 lilianesilvaduraes.pdf: 4940502 bytes, checksum: aad2827698668df37140c043979a25c4 (MD5) / Made available in DSpace on 2015-12-01T10:18:13Z (GMT). No. of bitstreams: 1 lilianesilvaduraes.pdf: 4940502 bytes, checksum: aad2827698668df37140c043979a25c4 (MD5) Previous issue date: 2015-02-27 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O município mineiro de Juiz de Fora se destaca no contexto epidemiológico por possuir indicadores de vulnerabilidade, como alto número de notificações do estado, com casos ocorrentes no perímetro urbano, que detém elevada densidade populacional, ocupação de áreas limítrofes com fragmento de mata, as quais margeiam áreas confirmadamente endêmicas, possibilitando o intercâmbio de artrópodes vetores com mamíferos hospedeiros. Tais áreas propiciam o desenvolvimento e a manutenção dos ciclos enzoótico e epidêmico para Febre Maculosa. Mediante ao exposto, o objetivo geral deste projeto foi investigar e mapear a relação natural entre espécies de Rickettsia sp. e seus potenciais vetores em cinco regiões de vulnerabilidade do município. Especificamente, objetivaram-se: a determinação da composição da fauna de ectoparasitos potencialmente vetores coletados sobre hospedeiros vertebrados, bem como livres no ambiente; análise do ciclo enzoótico através da detecção de rickettsias por meio de Reação em Cadeia da Polimerase (PCR); a caracterização dos agentes rickettsias por sequenciamento e análises filogenéticas e a avaliação dos aspectos epidemiológicos da rickettsias circulantes, dos vetores e seus hospedeiros vertebrados. Os ectoparasitos foram coletados no período de 2013-2014 em localidades que carecem de informações sobre a manutenção do ciclo de transmissão de rickettsias: Caetés, Igrejinha, Paula Lima, Torreões e Valadares. No total foram coletados e identificados 12.692 ectoparasitos, entre pulgas do gênero Ctenocephalides e carrapatos dos gêneros Amblyomma, Rhipicephalus e Dermacentor, com posterior processamento de 2.379 espécimes de maneira individual ou reunidos em lotes de acordo com localidade, sexo, estádio de desenvolvimento, constituindo 249 amostras. O DNA genômico destas amostras foi extraído e submetido à pesquisa de genes rickettsiais gltA (gênero Rickettsia) e ompA (Grupo da Febre Maculosa-GFM). Foram obtidas 29 amostras positivas, exceto para Caetés; sendo 16 para o gene gltA, com 11 amostras sequenciáveis, e 13 amostras para o gene ompA, com sete sequenciáveis; com destaque para Amblyomma cajennense sendo a espécie mais prevalente e com 4,29% de taxa de infecção mínima. Sequências com 99 e 100% de identidade com a espécie patogênica Rickettsia felis foram diagnosticadas em Ctenocephalides canis e Ctenocephalides felis. E de forma inédita o registro de Rickettsia bellii em Amblyomma dubitatum na Microrregião de Juiz de Fora, apresentando sequências com 97% de homologia. O presente trabalho revela uma maior distribuição geográfica de rickettsias do GFM, incluindo R. felis causadora da rickettsiose felis, na Microrregião de Juiz de Fora. / The city of Juiz de Fora, in the Brazilian state of Minas Gerais, stands out in the epidemiologic context for possessing vulnerability indicators, such as a high number of this state´s notifications, with cases occurring in the urban perimeter, which contains an elevated population density, and the occupation of limit areas with forests´ fragments, which have a frontier with confirmed endemic areas, making possible the interchange of arthropod vectors with mammal hosts. Such areas propitiate the development and the maintenance of the enzootic and the epidemic cycles for the Spotted Fever. By the above, the general objective of this project was to investigate and to map the natural relation between Rickettsia sp. Species and its potential vectors in five regions of vulnerability in the city. Specifically, here are the objectives: the determination of the fauna composition for ectoparasites potentially vectors, collected on vertebrate hosts, as well as loose in the environment; analysis of the enzootic cycle through the detection of rickettsias by Polymerase Chain Reaction (PCR); the rickettsias agents characterization through sequencing and phylogenetic analyses and the evaluation of the epidemiologic aspects of the circulating rickettsias, of the vectors and their vertebrate hosts. The ectoparasites were collected in the period of 2013-2014 in places that need information on the maintenance of rickettsias transmission cycle: Caetés, Igrejinha, Paula Lima, Torreões e Valadares. As a total, there were collected and identified 12,692 ectoparasites, among fleas from the gender Ctenocephalides and ticks from the genders Amblyomma, Rhipicephalus and Dermacentor, with a posterior processing of 2,379 specimens in an individual way or gathered in lots according to place, sex, development state, constituting 249 samples. The genomic DNA from these samples was extracted and submitted to research of rickettsiais gltA (gênero Rickettsia) e ompA (Grupo da Febre Maculosa-GFM) genes. There were obtained 29 positive samples, except for Caetés; from which 16 for the gltA gene, with 11 sequenced samples, and 13 samples for the ompA gene, with seven sequenced; especially Amblyomma cajennense as the most prevalent species and with 4.29% of minimal infection rate. Sequences with 99 and 100% of identity with the pathogenic species Rickettsia felis were diagnosed in Ctenocephalides canis and Ctenocephalides felis. And in a surprising way the register of Rickettsia bellii in Amblyomma dubitatum in Juiz de Fora´s Micro-region, presenting sequences with 97% of homology. This paper reveals a bigger geographic distribution of rickettsias from GFM, including R. felis that causes rickettsiosis felis, in Juiz de Fora´s Micro-region.

Submitted by Renata Lopes (renatasil82@gmail.com) on 2015-12-03T09:53:31Z No. of bitstreams: 1 lilianesilvaduraes.pdf: 4940502 bytes, checksum: aad2827698668df37140c043979a25c4 (MD5) / Rejected by Adriana Oliveira (adriana.oliveira@ufjf.edu.br), reason: Título: somente primeira letra da primeira palavra em maiúsculo e nomes próprios também. Palavras-chave: primeira letra de cada palavra em maiúsculo. Acrescentar uma em cada linha (não precisa colocar ponto e nem vírgula no final) on 2015-12-03T11:59:21Z (GMT) / Submitted by Renata Lopes (renatasil82@gmail.com) on 2015-12-03T12:48:28Z No. of bitstreams: 1 lilianesilvaduraes.pdf: 4940502 bytes, checksum: aad2827698668df37140c043979a25c4 (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2015-12-03T13:57:54Z (GMT) No. of bitstreams: 1 lilianesilvaduraes.pdf: 4940502 bytes, checksum: aad2827698668df37140c043979a25c4 (MD5) / Made available in DSpace on 2015-12-03T13:57:54Z (GMT). No. of bitstreams: 1 lilianesilvaduraes.pdf: 4940502 bytes, checksum: aad2827698668df37140c043979a25c4 (MD5) Previous issue date: 2015-02-27 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O município mineiro de Juiz de Fora se destaca no contexto epidemiológico por possuir indicadores de vulnerabilidade, como alto número de notificações do estado, com casos ocorrentes no perímetro urbano, que detém elevada densidade populacional, ocupação de áreas limítrofes com fragmento de mata, as quais margeiam áreas confirmadamente endêmicas, possibilitando o intercâmbio de artrópodes vetores com mamíferos hospedeiros. Tais áreas propiciam o desenvolvimento e a manutenção dos ciclos enzoótico e epidêmico para Febre Maculosa. Mediante ao exposto, o objetivo geral deste projeto foi investigar e mapear a relação natural entre espécies de Rickettsia sp. e seus potenciais vetores em cinco regiões de vulnerabilidade do município. Especificamente, objetivaram-se: a determinação da composição da fauna de ectoparasitos potencialmente vetores coletados sobre hospedeiros vertebrados, bem como livres no ambiente; análise do ciclo enzoótico através da detecção de rickettsias por meio de Reação em Cadeia da Polimerase (PCR); a caracterização dos agentes rickettsias por sequenciamento e análises filogenéticas e a avaliação dos aspectos epidemiológicos da rickettsias circulantes, dos vetores e seus hospedeiros vertebrados. Os ectoparasitos foram coletados no período de 2013-2014 em localidades que carecem de informações sobre a manutenção do ciclo de transmissão de rickettsias: Caetés, Igrejinha, Paula Lima, Torreões e Valadares. No total foram coletados e identificados 12.692 ectoparasitos, entre pulgas do gênero Ctenocephalides e carrapatos dos gêneros Amblyomma, Rhipicephalus e Dermacentor, com posterior processamento de 2.379 espécimes de maneira individual ou reunidos em lotes de acordo com localidade, sexo, estádio de desenvolvimento, constituindo 249 amostras. O DNA genômico destas amostras foi extraído e submetido à pesquisa de genes rickettsiais gltA (gênero Rickettsia) e ompA (Grupo da Febre Maculosa-GFM). Foram obtidas 29 amostras positivas, exceto para Caetés; sendo 16 para o gene gltA, com 11 amostras sequenciáveis, e 13 amostras para o gene ompA, com sete sequenciáveis; com destaque para Amblyomma cajennense sendo a espécie mais prevalente e com 4,29% de taxa de infecção mínima. Sequências com 99 e 100% de identidade com a espécie patogênica Rickettsia felis foram diagnosticadas em Ctenocephalides canis e Ctenocephalides felis. E de forma inédita o registro de Rickettsia bellii em Amblyomma dubitatum na Microrregião de Juiz de Fora, apresentando sequências com 97% de homologia. O presente trabalho revela uma maior distribuição geográfica de rickettsias do GFM, incluindo R. felis causadora da rickettsiose felis, na Microrregião de Juiz de Fora. / The city of Juiz de Fora, in the Brazilian state of Minas Gerais, stands out in the epidemiologic context for possessing vulnerability indicators, such as a high number of this state´s notifications, with cases occurring in the urban perimeter, which contains an elevated population density, and the occupation of limit areas with forests´ fragments, which have a frontier with confirmed endemic areas, making possible the interchange of arthropod vectors with mammal hosts. Such areas propitiate the development and the maintenance of the enzootic and the epidemic cycles for the Spotted Fever. By the above, the general objective of this project was to investigate and to map the natural relation between Rickettsia sp. Species and its potential vectors in five regions of vulnerability in the city. Specifically, here are the objectives: the determination of the fauna composition for ectoparasites potentially vectors, collected on vertebrate hosts, as well as loose in the environment; analysis of the enzootic cycle through the detection of rickettsias by Polymerase Chain Reaction (PCR); the rickettsias agents characterization through sequencing and phylogenetic analyses and the evaluation of the epidemiologic aspects of the circulating rickettsias, of the vectors and their vertebrate hosts. The ectoparasites were collected in the period of 2013-2014 in places that need information on the maintenance of rickettsias transmission cycle: Caetés, Igrejinha, Paula Lima, Torreões e Valadares. As a total, there were collected and identified 12,692 ectoparasites, among fleas from the gender Ctenocephalides and ticks from the genders Amblyomma, Rhipicephalus and Dermacentor, with a posterior processing of 2,379 specimens in an individual way or gathered in lots according to place, sex, development state, constituting 249 samples. The genomic DNA from these samples was extracted and submitted to research of rickettsiais gltA (gênero Rickettsia) e ompA (Grupo da Febre Maculosa-GFM) genes. There were obtained 29 positive samples, except for Caetés; from which 16 for the gltA gene, with 11 sequenced samples, and 13 samples for the ompA gene, with seven sequenced; especially Amblyomma cajennense as the most prevalent species and with 4.29% of minimal infection rate. Sequences with 99 and 100% of identity with the pathogenic species Rickettsia felis were diagnosed in Ctenocephalides canis and Ctenocephalides felis. And in a surprising way the register of Rickettsia bellii in Amblyomma dubitatum in Juiz de Fora´s Micro-region, presenting sequences with 97% of homology. This paper reveals a bigger geographic distribution of rickettsias from GFM, including R. felis that causes rickettsiosis felis, in Juiz de Fora´s Micro-region. Keywords:

Rodents have been reported to play a significant role as reservoirs of over 22 rodent-associated Bartonella species. In this study, we contrast prevalence and diversity of Bartonella infections in 377 small mammals, representative of three terrestrial rodent genera, namely Aethomys, Gerbilliscus and Rhabdomys and one subterranean mole-rat species (Bathyergus suillus). The latter was sampled in close proximity to an informal human settlement, whereas the afore-mentioned murid rodent genera were sampled across a range of landscapes inclusive of natural, agricultural, urban, peri-urban and rural settings, from three provinces (Free State, Gauteng and Western Cape) in South Africa. Molecular estimates of Bartonella infection rates were determined through multi-gene screening of DNA extracted from clinical samples, primarily heart and spleen. PCR assays targeting the citrate synthase (gltA) and NADH dehydrogenase gamma subunit (nuoG) and/or beta subunit of bacterial RNA polymerase (rpoB) genes were used to ensure enhanced molecular estimates of Bartonella prevalence. Aethomys had the highest infection rate (86.7%), whereas Rhabdomys had the lowest (15%). Nucleotide sequencing and phylogenetic analyses revealed that the different primers sets used for Bartonella screening have different affinities to the different strains present in rodents from South Africa. Furthermore, the presence of Bartonella co-infections, confirmed through the presence of multiple peaks at 15% of the nucleotide sequences sites, ranged from 33.8% (in Aethomys) to 42.9% (in Gerbilliscus species) for the gltA gene region. For Aethomys ineptus, of the discrete Bartonella lineages recovered, one was closely related to zoonotic B. elizabethae. The latter species, which is associated with Rattus hosts worldwide and has been linked to cases of human endocarditis, suggests spillover from invasive to indigenous rodents. This is supported by previous studies indicating that indigenous Micaelamys namaquensis, a highly adaptable species, which like Aethomys is capable of utilizing natural and modified landscapes also hosts B. elizabethae-related lineages. Of potential public health importance, Bathyergus suillus were shown to be infected with a zoonotic Bartonella species, B. rochalimae. Our results further indicate that the level of anthropogenic transformation is significantly correlated with Bartonella prevalence, with Rhabdomys sampled from rural settings in the Western Cape Province having infection rates of 36% versus 0% in a nearby urban setting. This study also uncovered high levels of strain diversity in members of the Gerbilliscus cryptic species complex, sampled from an agricultural setting. The overall Bartonella PCR-positivity rate was 67.5 % and the gltA gene phylogeny confirmed the presence of six discrete Bartonella gerbil-specific lineages (I-VI). Lineages I and II clustered with Bartonella strains identified previously in G. leucogaster sampled from Sandveld nature reserves in the Free State Province South Africa, whereas lineages III-VI comprised of lineages that were restricted to either G. leucogaster or to G. brantsii, indicative of host-specificity. From the findings of this study, it is clear that the public health importance of the Bartonella species present in indigenous rodents warrants further investigation as at least two species, with known zoonotic potential (B. elizabethae and B. rochalimae) were shown to be present in rodents sampled in close proximity to human settlements. / Thesis (PhD (Zoology))--University of Pretoria, 2019. / This research was made possible through the financial support from the (AZD-IRT), CDC Co-Ag 5 NU2GGH001874-02-00 and through the NRF incentive and SARChI PI funding awarded to ADSB. / Zoology and Entomology / PhD (Zoology) / Unrestricted