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The genus Neorickettsia comprises trematode-associated bacteria that can cause diseases in animals and humans. Despite detection of Neorickettsia antigens in the intestine of coatis kept in captivity in southern Brazil through immunohistochemistry, the molecular identity of the bacteria in South American procyonids remains elusive. The aim of the present study was to investigate the occurrence of Neorickettsia sp. in blood samples from coatis in central-western Brazil. Between March 2018 and January 2019, animals were captured and recaptured in two areas of the Cerrado (Parque Estadual do Prosa, PEP; and Vila da Base Aérea, VBA) located in the city of Campo Grande, state of Mato Grosso do Sul, central-western Brazil. All captures were performed according to convenience. DNA from 97 blood samples was subjected to nested PCR (nPCR) targeting a fragment of the 16S rRNA gene of Neorickettsia sp. Six samples (3.6%; five from VBA and one from PEP) from different coatis were positive in nPCR based on the 16S rRNA. The sequences obtained (~500 bp) showed ˃ 99% similarity to N. risticii. Phylogenetic analysis clustered the sequences detected in the present study in a clade with N. risticii. This is the first molecular detection of Neorickettsia sp. in coatis in Brazil. Resumo O gênero Neorickettsia compreende bactérias associadas a trematódeos que podem causar doenças em animais e humanos. Apesar da detecção de antígenos de Neorickettsia por imuno-histoquímica no intestino de quatis mantidos em cativeiro no sul do Brasil, a identidade molecular da bactéria em procionídeos da América do Sul permanece indefinida. O presente trabalho teve como objetivo investigar a ocorrência de Neorickettsia sp. em amostras de sangue de quatis do Centro-Oeste do Brasil. Entre março de 2018 e janeiro de 2019, os animais foram capturados em duas áreas de Cerrado (Parque Estadual do Prosa PEP e Vila da Base Aérea VBA) localizadas na cidade de Campo Grande, Mato Grosso do Sul, Centro-Oeste do Brasil. Todas as capturas e recapturas foram realizadas por conveniência. O DNA de 97 amostras de sangue foi submetido a "nested" (nPCR), baseada em um fragmento do gene 16S rRNA de Neorickettsia sp. Seis (3,6% - 5 de VBA e 1 de PEP) amostras de quatis diferentes foram positivas na nPCR, baseada no rRNA 16S. As sequências obtidas (~500 pb) apresentaram ˃99% de identidade com N. risticii. A inferência filogenética agrupou as sequencias detectadas no presente estudo em um clado com N. risticii. Esta é a primeira detecção molecular de Neorickettsia sp. em quatis do Brasil.

A free-living, adult male maned wolf (Chrysocyon brachyurus) was referred to the Governador “Laudo Natel” – FCAV/Unesp veterinary hospital after being found with skin lesions and a fracture on the right pelvic limb, which had to be amputated due to compromised integrity. Around 20 days later, bilateral accentuated swollen on humerus-radius-ulna articulation was observed. The synovial liquid was drained and sent to the laboratory for synovial cytology with Rosenfeld staining that revealed predominantly degenerated neutrophils with karyolytic chromatin associated with intracellular inclusions suggestive of Hepatozoon sp. gametocytes. Blood and synovial liquid samples were submitted to molecular analysis, aiming to amplify the Hepatozoon spp. 18S rRNA gene fragment. Despite the positioning of the found Hepatozoon sequence together with Hepatozoon canis previously detected in domestic carnivores, the BLAST analysis showed only 98% identity with H. canis. To the best of the authors’ knowledge, this is the first time a Hepatozoon was detected in the synovial liquid by clinical pathology and molecular analyses. Resumo Um lobo guará (Chrysocyon brachyurus) adulto, macho, de vida livre foi encaminhado para atendimento no hospital veterinário Governador “Laudo Natel” – FCAV/Unesp após ser encontrado com lesões de pele e fratura em membro pélvico direito, sendo amputado devido a comprometimento da integridade do membro. Aproximadamente 20 dias após a chegada ao hospital, foi notado acentuado aumento de volume bilateral em região de articulação úmero-rádio-ulnar. O líquido sinovial foi drenado e enviado para análise citológica com coloração de Rosenfeld, revelando a presença de neutrófilos degenerados com cromatina cariolítica associados a inclusões intracelulares sugestivas de gametócitos de Hepatozoon sp. Amostras de sangue e líquido sinovial foram submetidas a análises moleculares visando amplificar um fragmento do gene 18S rRNA de Hepatozoon spp. Apesar da sequência de Hepatozoon detectada se posicionar filogeneticamente no mesmo clado que H. canis previamente detectado em carnívoros domésticos, o resultado da análise do BLAST mostrou somente 98% de identidade com H. canis. De acordo com o conhecimento dos autores, esta é a primeira vez que Hepatozoon foi detectado no líquido sinovial por meio de patologia clínica e análises moleculares.

Submitted by Livia Perles (liviaperles@hotmail.com) on 2019-03-22T13:15:03Z No. of bitstreams: 1 Dissertação_Lívia_Perles.pdf: 2901269 bytes, checksum: ba9042d31388702f6e9dabbfc6d5c252 (MD5) Rejected by Tatiana Camila Gricio (tatiana.gricio@unesp.br), reason: Solicitamos que realize a correção na submissão seguindo as orientações abaixo: 1 - Segundo a CAPES (Portaria nº 206, de 4 de setembro de 2018), o agradecimento deve conter exatamente a seguinte redação: O presente trabalho foi realizado com apoio da Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) - Código de Financiamento 001. Agradecemos a compreensão. on 2019-03-22T18:29:59Z (GMT) Submitted by Livia Perles (liviaperles@hotmail.com) on 2019-03-24T15:04:25Z No. of bitstreams: 1 Dissertação Lívia 24-03-19.pdf: 2777459 bytes, checksum: 61618e805d66b9c5a15905de8e7f7889 (MD5) Approved for entry into archive by Tatiana Camila Gricio (tatiana.gricio@unesp.br) on 2019-03-25T13:50:54Z (GMT) No. of bitstreams: 1 perles_l_me_jabo_par.pdf: 815334 bytes, checksum: 932643488c12aea5138cfd010e5be6b9 (MD5) Made available in DSpace on 2019-03-25T13:50:54Z (GMT). No. of bitstreams: 1 perles_l_me_jabo_par.pdf: 815334 bytes, checksum: 932643488c12aea5138cfd010e5be6b9 (MD5) Previous issue date: 2019-02-27 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) O gênero Hepatozoon spp. engloba protozoários parasitas de uma ampla variedade de vertebrados terrestres. Recentemente, vem se investigando o possível papel de roedores como hospedeiros intermediários e paratênicos nos ciclos de transmissão de espécies de Hepatozoon parasitas de carnívoros domésticos e selvagens. O presente estudo teve como objetivo investigar a presença e caracterizar o DNA de Hepatozoon spp. em amostras de baço de 31 gêneros de roedores amostrados em cinco biomas brasileiros. Das 462 amostras analisadas 195 amostras foram positivas em um ou ambos os protocolos utilizados. Hepatoozon spp. foi detectado em 24 gêneros de roedores, com primeiro relato da infecção pelo referido parasita em Rattus rattus, Mus musculus, Proechimys roberti, P. cuvieri, G. spixii, Hylaeamys megacephalus, Gracilinanus agilis, Cerradomys scotti, C. akroai, C. marinhus e Wiedomys cerradensis. As análises Bayesiana e de Máxima Verossimilhança das sequências 18S rRNA obtidas mostraram a presença de três clados de Hepatozoon, sendo um clado composto por sequências de Hepatozoon sp. detectados em roedores e répteis (cobras, jacarés e lagartos), um clado composto por Hepatozoon americanum e espécies de Hepatozoon detectadas em canídeos e felídeos selvagens, e um clado agrupando Hepatozoon canis e Hepatozoon spp. detectados em canídeos domésticos e selvagens e Rhipicephalus sanguineus. As sequências de Hepatozoon do presente estudo foram agrupadas no clado de roedores e répteis. A análise de distância pelo software Splitstree revelou que as sequências do presente estudo foram agrupadas em um grupo com sequências de Hepatozoon de outros roedores do Brasil e do mundo, próximas às sequências de Hepatozoon detectadas em répteis. As sequências de Hepatozoon obtidas de felídeos e canídeos formaram grupos distintos daquelas detectadas em roedores. Para análise de haplótipos 18S rRNA de Hepatozoon spp. detectados em roedores em outros estudos realizados no Brasil até o presente momento foram escolhidas 26 sequências e a análise foi realizada com o software TCS. Seis haplótipos foram encontrados, sendo o haplótipo 1 o mais frequente. Os resultados do presente estudo sugerem a existência de estruturação genética das espécies de Hepatozoon que ocorrem em roedores no Brasil de acordo com a localidade geográfica. The Hepatozoon spp. genus includes protozoan parasites of a wide range of terrestrial vertebrates. Recently, the role of rodents as intermediate and paratenic hosts in the transmission of Hepatozoon species parasites of domestic and wild carnivores, has been investigated. The present study aimed to investigate the presence and characterization of Hepatozoon spp. in spleen samples of 31 genera of rodents sampled in five Brazilian biomes. Out of 462 samples analyzed, 195 were positive in one or in both protocols. Hepatoozon spp. was detected in 24 genera of rodents, with the first report of the infection in Rattus rattus, Mus musculus, Proechimys roberti, P. cuvieri, G. spixii, Hylaeamys megacephalus, Gracilinanus agilis, Cerradomys scotti, C. akroai, C. marinhus and Wiedomys cerradensis. Bayesian and Maximum Likelihood analyzes of the obtained 18S rRNA sequences demonstrated the presence of three clades of Hepatozoon, one clade being composed of Hepatozoon sp. detected in rodents and reptiles (snakes, alligators and lizards), a clade composed of Hepatozoon americanum and Hepatozoon species detected in canids and wild felids, and a clade grouping Hepatozoon canis and Hepatozoon spp. detected in domestic and wild canids and in Rhipicephalus sanguineus. Hepatozoon sequences of the present study were grouped in the clade of rodents and reptiles. Distance analysis by Splitstree software revealed that the sequences of the present study were grouped into a group with Hepatozoon sequences from other rodents from Brazil and the world, close to the Hepatozoon sequences detected in reptiles. For analysis of 18S rRNA haplotypes of Hepatozoon spp. detected in rodents from other studies conducted in Brazil up to the moment, 26 sequences were chosen and the analysis was performed with the TCS software. Six haplotypes were found, with haplotype 1 being the most frequent. Results of the present study suggest the existence of genetic structuring of Hepatozoon species in rodents in Brazil according to the geographical location. FAPESP: 2015/14896-1 CNPq: 2014/401.403.120.16-5

Made available in DSpace on 2019-10-06T16:34:19Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-01-01 Yellow fever (YF) is a zoonotic arthropod-borne disease that is caused by the yellow fever virus (YFV) and characterized by a sylvatic and urban cycle. Its most severe presentation is manifested as a hemorrhagic disease, and it has been responsible for thousands of deaths in the last decades. This study describes the public health approaches taken to control the 2016-2017 YF outbreak in nonhuman primates (NHPs) that took place in the northeastern region of São Paulo state, Brazil. NHPs recovered from the field were necropsied, and YF diagnoses were made at the Laboratory of Molecular Virology, Ribeirão Preto Medical School and the Center of Pathology, Adolfo Lutz Institute of São Paulo. NHP samples were inoculated into Vero cells for YFV isolation. RNA extraction was performed directly from NHP tissues and tested by RT-qPCR. YFV-positive samples were confirmed by sequencing. Based on the rapid RT-qPCR results, surveillance actions were implemented in the entire region. Confirmatory histopathology and immunohistochemistry for YFV were also performed. Among nine NHPs, gross hepatic involvement was observed in six animals, five of which were YFV-RT-qPCR-positive. One YFV was isolated from the serum of an infant NHP. YFV RNA sequences diverged from the virus responsible for the last epizootic that occurred in São Paulo state, but it was similar to the current Brazilian epizootic. Public health actions included dissemination of information on YF transmission, investigation of the probable location of NHP infection, characterization of the environment, and subsequent creation of the blueprint from which prevention and control measures were implemented. The YFV sylvatic cycle occurred in the periurban areas of the northeastern region of São Paulo state, but no human cases were reported during this period, showing that integrated actions between human, animal, and environmental health professionals were critical to restrain the virus to the sylvatic cycle. Department of Internal Medicine Ribeirão Preto Medical School University of São Paulo FMRP/USP, Av. Bandeirantes, 3900, Vila Monte Alegre Veterinary Preventive Medicine and Animal Reproduction Department Faculty of Agricultural and Veterinary Sciences Universidade Estadual Paulista Júlio de Mesquita Filho (FCAV/Unesp), Access Way Prof. Paulo Donato Castellane s/n Animal Pathology Department Faculty of Agricultural and Veterinary Sciences Universidade Estadual Paulista Júlio de Mesquita Filho (FCAV/Unesp), Access Way Prof. Paulo Donato Castellane s/n Municipal Health Secretary, Av. General Glicério, 569, Center Nucleus of Pathological Anatomy Pathology Center Adolfo Lutz Institute of São Paulo State Secretary of Health, Av. Dr. Arnaldo, 351 Veterinary Preventive Medicine and Animal Reproduction Department Faculty of Agricultural and Veterinary Sciences Universidade Estadual Paulista Júlio de Mesquita Filho (FCAV/Unesp), Access Way Prof. Paulo Donato Castellane s/n Animal Pathology Department Faculty of Agricultural and Veterinary Sciences Universidade Estadual Paulista Júlio de Mesquita Filho (FCAV/Unesp), Access Way Prof. Paulo Donato Castellane s/n

Hepatozoon species are the most common intracellular hemoparasite found in reptiles. Hepatozoon caimani, whose vectors are Culex mosquitoes, has been detected in a high prevalence among caimans in Brazil by blood smears examinations. The present work aimed to detect and characterize the Hepatozoon spp. found in 33 caimans (24 free-ranging and 9 captive; 28 males and 5 females) (Caiman crocodilus yacare) sampled at Poconé, North Pantanal, state of Mato Grosso, Brazil, using blood smears examinations and molecular techniques. Hepatozoon spp.-gametocytes were found in 70.8% (17/24) and 88.8% (8/9) of blood smears from free-ranging and captive caimans, respectively. Hepatozoon spp. 18S rRNA DNA was found in 79.2% (19/24) and 88.8% (8/9) of free-ranging and captive caimans, respectively. Comparative analysis of parasitized and non-parasitized erythrocytes showed that all analyzed features were significantly different (P