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Made available in DSpace on 2022-04-29T01:26:52Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-10-19 Serum samples from 714 equids of Itaguaí and Serrana microregions, Rio de Janeiro, southeastern Brazil, were examined by indirect fluorescent antibody test (titer 1:80) for Theileria equi. The prevalence in the microregions and factors associated with seropositivity were evaluated and the prevalence ratio (PR) calculated. The overall prevalence of T. equi infection was 81.09% (n = 579), with higher prevalence (p < 0.05) in the Itaguaí (85.43%) when compared to Serrana microregion (76.92%). The geographic area, altitude, farming condition and area of origin of equids were associated (p < 0.05) with seropositivity for T. equi. Equids reared in the Itaguaí microregion (PR = 1.11, p = 0.003) and at altitudes below 500 m (PR = 1.10; p = 0,014) were more likely to be seropositive for T. equi. Furthermore, when equids were born in the farm (PR = 1.10, p = 0.008) and reared with poor farming conditions (PR = 1.13, p = 0.018) they were more likely to be exposed to T. equi. The main ticks found on equids were Amblyomma cajennense and Dermacentor (Anocentor) nitens. The microregions studied are endemic areas for equine theileriosis and there exists enzootic stability for T. equi. Only factors related to the collection area of serum samples influenced the seropositivity of equids for T. equi in that region. Universidade Federal Rural do Rio de Janeiro UFRRJ Departamento de Medicina e Cirurgia Veterinária Instituto de Veterinária Universidade Federal Rural do Rio de Janeiro - UFRRJ Departamento de Patologia Veterinária Faculdade de Ciências Agrárias e Veterinárias Universidade Estadual Paulista - UNESP Departamento de Parasitologia Animal Instituto de Veterinária Universidade Federal Rural do Rio de Janeiro - UFRRJ Departamento de Patologia Veterinária Faculdade de Ciências Agrárias e Veterinárias Universidade Estadual Paulista - UNESP

Made available in DSpace on 2022-04-29T02:17:14Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-10-01 Neosporosis and toxoplasmosis are parasitic diseases which can cause reproductive problems in goats and sheep. The current study aimed to determine the occurrence of anti-Neospora caninum and anti-Toxoplasma gondii IgG antibodies in goats and sheep from the districts of Amarante do Maranhão and Buritirana, Imperatriz microregion, western area of Maranhão State, northeastern Brazil, and to assess factors associated to infection by these etiologic agents. Blood samples from 110 animals (46 goats and 64 sheep) from five herds were collected, and indirect immunofluorescence assay was used for serological testing. Of 46 goat samples, 17.39% (n = 8) showed anti-N. caninum antibodies and 4.35% (n = 2) anti-T. gondii, while of 64 sheep samples 4.69% (n = 3) and 18.75% (n = 12) showed anti-N. caninum and anti-T. gondii antibodies, respectively. No significant difference regarding the presence of domestic cats and/or dogs on the property and veterinary care was seen for both etiologic agents studied. However, food supplementation and animal reproductive failure were significantly (p < 0.05) for N. caninum among sheep and goats, respectively. The current study showed that goats and sheep in western Maranhão are exposed to N. caninum and T. gondii. It is the first evidence of these agents in small ruminants in this region. Departamento de Medicina e Cirurgia Veterinária - DMCV Instituto de Veterinária Universidade Federal Rural do Rio de Janeiro - UFRRJ Departamento de Parasitologia Animal - DPA Instituto de Veterinária Universidade Federal Rural do Rio de Janeiro - UFRRJ Escola de Medicina Veterinária e Zootecnia Universidade Federal do Tocantins - UFT Departamento de Patologia Veterinária Faculdade de Ciências Agrárias e Veterinárias - FCAV Universidade Estadual Paulista - UNESP Departamento de Patologia Veterinária Faculdade de Ciências Agrárias e Veterinárias - FCAV Universidade Estadual Paulista - UNESP

Submitted by Sandra Pereira (srpereira@ufrrj.br) on 2020-08-10T19:44:27Z No. of bitstreams: 1 2011 - Huarrisson Azevedo Santos.pdf: 1223931 bytes, checksum: 542b1c4c1fd9100a410f1fe8a7772c37 (MD5) Made available in DSpace on 2020-08-10T19:44:27Z (GMT). No. of bitstreams: 1 2011 - Huarrisson Azevedo Santos.pdf: 1223931 bytes, checksum: 542b1c4c1fd9100a410f1fe8a7772c37 (MD5) Previous issue date: 2011-04-11 Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior, CAPES, Brasil. Anaplasma phagocytophilum is an emerging pathogen of humans, horses, and dogs worldwide that is transmitted by Ixodid ticks and maintained in a variety of small wild mammal species.The objective of the present study was to evaluate the presence of A. phagocytophilum DNA in dogs and ticks of Itaguai microregion, Rio de Janeiro, as well as establish the associated factors (age, gender, breed, tick infestation, etc) and possible hematological alterations with A. phagocytophilum infection in dogs. Anaplasma phagocytophilum was detected for the first time from Brazil, in dogs the ticks of urban the rural areas in the Itaguai microregion, Rio de Janeiro state, by real-time polymerase chain reaction (qPCR). Of 398 samples, 24 (6.03%) were positive, with a CT ranging from 26 to 35 cycles. Amplicons from one positive sample were cloned in the pGEM-T and sequenced. The sequence obtained demonstrated 100% identity with other A. phagocytophilum sequences published in the GenBank database. The analytical sensitivity of RT-PCR using SYBR green system was able to detect three plasmid copies when defined numbers of plasmid copies containing 122 base pairs from the msp2 gene were used. The assay was considered specific when DNA from bacteria closely related to A. phagocytophilum was placed in the reaction. A fragment of 122pb msp2 gene of A. phagocytophilum was detected by TaqMan-PCR in one female tick of Amblyomma cajennense and in five Rhipicephalus sanguineus, being four males and one female. Positive tick samples obtained CT that ranged from 26.75 to 33.14 cycles. The TaqMan-PCR assay was capable to detect a single copy of A. phagocytophilum msp2 gene in DNA tick sample. Tick infestation was observed in 66.67% (n = 16) of PCR positive dogs. The tick species identified in positive dogs were R. sanguineus and A. cajennense, with predominance of the latter tick species. The history of tick infestation, access to forest areas and environment condition of cleanliness were factors associated with A. phagocytophilum infection in dogs. The main hematologic findings in dogs infected with A. phagocytophilum were thrombocytopenia, lymphopenia and increased mean platelet volume.These results demonstrate that the canine granulocytic anaplasmosis agent is present in regions in which dogs could be a source of infection for tick vectors. This is the first report of A. phagocytophilum in Brazil, a emergent zoonotic agent responsible of Human Granulocitic Anaplasmosis. Anaplasma phagocytophilum ? um pat?geno emergente capaz de infectar humanos, equinos e c?es em todo o mundo. Essa bact?ria ? transmitida por carrapatos ixod?deos e mantida na natureza por uma variedade de pequenos mam?feros silvestres. O objetivo do presente estudo foi avaliar a presen?a do DNA de A. phagocytophilum em c?es e carrapatos da microrregi?o de Itagua?, Rio de Janeiro, al?m de estabelecer os fatores associados (idade, sexo, ra?a, infesta??o por carrapatos, etc) e as poss?veis altera??es hematol?gicas com a infec??o por A. phagocytophilum em c?es desta regi?o. Essa bact?ria foi detectada pela primeira vez no Brasil, em c?es e carrapatos de ?reas rurais e urbanas da microrregi?o de Itaguai, RJ atrav?s da Rea??o em Cadeia da Polimerase (PCR) em tempo real. Dos 398 c?es amostrados, 24 (6,03%) foram considerados positivos, com limite de detec??o (CT) variando entre 26 a 35 ciclos. As amostras positivas clonadas em pGEM-T apresentaram 100% de identidade com outras sequ?ncias de A. phagocytophilum depositadas no GenBank. Utilizando n?meros definidos de c?pias do plasm?dio contendo um inserto de 122pb do gene msp2 a sensibilidade anal?tica do ensaio de PCR em tempo real atrav?s do sistema SYBR green foi capaz de detectar at? tr?s c?pias do plasm?dio. O ensaio foi considerado espec?fico quando DNA de bact?rias proximamente relacionadas ? A. phagocytophilum foi colocado na rea??o. Um fragmento de 122pb do gene msp2 de A. phagocytophilum foi detectado atrav?s da TaqMan- PCR em um carrapato f?mea da esp?cie Amblyomma cajennense e em cinco Rhipicephalus sanguineus, sendo quatro machos e uma f?mea. As amostras de carrapatos positivas obtiveram CT que variaram entre 26,75 e 33,14 ciclos. A t?cnica de PCR em tempo real utilizando o sistema TaqMan foi capaz de detectar uma ?nica c?pia do gene msp2 de A. phagocytophilum em amostras de DNA de carrapatos. Infesta??o por carrapatos foram observadas em 66,67% (n=16) dos c?es positivos na PCR. As esp?cies de carrapatos identificadas nos c?es positivos foram R. sanguineus e A. cajennense, com predomin?ncia desta ?ltima esp?cie de carrapato. O hist?rico de infesta??o por carrapatos, acesso a ?reas de mata e condi??o de limpeza do ambiente foram os fatores que apresentaram associa??o com a presen?a do DNA de A. phagocytophilum em c?es na regi?o estudada. Os principais achados hematol?gicos em c?es infectados com A. phagocytophilum foram trombocitopenia, linfopenia e aumento do volume plaquet?rio m?dio. Os resultados do presente estudo demonstram que o agente da Anaplasmose Granuloc?tica Canina circula na regi?o estudada e os c?es podem servir como fonte de infec??o para carrapatos vetores do agente. Este ? o primeiro relato de A. phagocytophilum no Brasil, um agente zoon?tico emergente respons?vel pela Anaplasmose Granuloc?tica Humana.

Este trabalho teve o objetivo de avaliar a reação de imunofluorescência indireta (RIFI), ensaio imunoenzimático (ELISA) e a reação de fixação do complemento (RFC) no diagnóstico de Theileria equi em amostras de soro de 79 equinos na Universidade Federal Rural do Rio de Janeiro (UFRRJ), Seropédica, RJ, Brasil. Houve reação positiva para Theileria equi em 74,7, 75,9 e 60,8% das amostras testadas pela RIFI, ELISA e RFC, respectivamente. Observou-se discrepância em 16,45% (n=13) das amostras de soro testadas pelo ELISA indireto e RIFI. Quando comparado a RIFI e a RFC, a discrepância observada entre os soros testados foi de 36,70% (n=29). O teste ELISA indireto e a RFC apresentaram discordância em 37,97% (n=30) das amostras de soros. Os resultados do presente estudo sugerem que a melhor alternativa para o diagnóstico sorológico de T. equi em eqüinos portadores é a associação dos testes de RIFI e ELISA indireto, especialmente para a realização de estudos soroepidemiológicos. This study was carried out to evaluate indirect fluorescent antibody test (IFAT), enzyme-linked immunosorbent assay (ELISA) and complement fixation test (CFT) of Theileria equi diagnosis in sera samples of 79 horses at Universidade Federal Rural do Rio de Janeiro (UFRRJ), Seropédica, RJ, Brazil were tested. Positive reaction was obtained in 74.7, 75.9 and 60.8% of samples tested by IFAT, indirect ELISA and CFT, respectively. Discrepancy was observed in 16.45% (n=13) of serum samples tested by ELISA and IFAT. While IFAT and CFT were compared, the discrepancy observed among the samples tested were 36.71% (n=29). Indirect ELISA and CFT test presented disagreement in 37.97% (n=30) of serum samples tested. Results of present study suggests that the best alternative for serological diagnosis T. equi in carriers horses is the combined use of IFAT and indirect ELISA test, especially for accomplishment of seroepidemiological studies.

Made available in DSpace on 2016-04-28T20:15:32Z (GMT). No. of bitstreams: 1 2009 - Huarrisson Azevedo Santos.pdf: 780165 bytes, checksum: de6d416bd5e351432ab7a787d05594f2 (MD5) Previous issue date: 2009-02-20 The objective of the present study was to describe the biological aspects of Argas (Persicargas) miniatus in controlled conditions to 27?1?C and 80?10% (RH) and outdoor condition. The eggs originated of A. (P.) miniatus females were incubated in conditions mentioned above. The emerged larvae were fed in chick, while the nymphal and adult stages were fed in chicken. The nymphal instars biological characteristics were studied to 15, 30 and 60 days fast periods. The biological aspects of nymphal and adult stages were studied in outdoor condition in the dry (May to October) and rainy (November to April) station. The feeding time, recovery percentage, mortality percentage, weight before and after feeding, molting periods, preoviposition and oviposition, daily eggs output, incubation period, hatching percentage, estimated reproduction and nutritional index were evaluated. The larvae medium weight was 0,94 ? 0,13mg with a medium weight gain of approximately 81,37 times. In acclimatized camera the molting medium period was 6,37?0,24 days, while in outdoor conditions it was 8,12?0,95 days. When evaluated the larvae fixation capacity submitted to different fast periods was observed that unfed larvae maintained in acclimatized camera and outdoor conditions were capable to attachment in birds with fast periods from 6 to 75 days and 8 to 60 days, respectively. When nymphs were submitted to 15 and 30 days fast periods, in both conditions, the development of second and third nymphal instars happened. When submitted to 60 fast period days were verified mortality of 28 and 37% in acclimatized camera and outdoor condition, respectively and survivors do not attachment on the hosts. The nymphs of second instar submitted to 60 fast days developed, in both conditions, in third nymphal instar and male adults. Still in the group submitted to 60 fast days, the nymphs of third instar developed in adults (42,42% and 40,54% males; 36,36% and 48,65% females in outdoor condition and acclimatized camera, respectively) and the others developed in third nymphs instars, and further, developed in female adults. The medium number of produced eggs varied of 46 to 138 eggs in the 18 female s oviposition maintained in controlled conditions; 41 to 108 eggs in the 9 female s oviposition at the rainy station; and 74 to 138 eggs for the females at the dry station, with significant difference in all experimental conditions. The medium duration of the cycle with adults occurrence in N2 was 49.05 days in acclimatized camera, 53.01 days in the dry station and 67.41 days in the rainy station. The results obtained in the present study demonstrate that the biological aspects of A. (P.) miniatus are influenced by climatic factors, occurring a biological cycle prolongation during the dry station, period that the temperature is lower. O objetivo do presente estudo foi descrever os aspectos biol?gicos de Argas (Persicargas) miniatus em condi??es controladas a 27?1?C e 80?10% (UR) e ambiente de laborat?rio. Os ovos oriundos de f?meas de A. (P.) miniatus foram incubados nas condi??es descritas acima. As larvas emergidas foram alimentadas em aves jovens, enquanto que os est?gios ninfais e adultos foram alimentados em aves adultas. As caracter?sticas biol?gicas dos ?nstares ninfais foram estudadas em per?odos de jejum de 15, 30 e 60 dias. Os aspectos biol?gicos dos est?gios ninfais e adultos foram estudados em condi??o ambiente de laborat?rio nas esta??es secas (Maio a Outubro) e chuvosas (Novembro a Abril). Foram avaliados: tempo de alimenta??o, taxa de recupera??o, taxa de mortalidade, peso antes e ap?s a alimenta??o, per?odos de muda, pr?-postura e postura, n?mero de posturas e de ovos, per?odo de incuba??o, percentual de eclos?o, reprodu??o estimada e ?ndice nutricional. O peso m?dio das larvas foi de 0,94 ? 0,13mg com ganho m?dio de peso de aproximadamente 81,37 vezes. Em c?mara climatizada o per?odo m?dio de muda foi de 6,37?0,24 dias, enquanto que em condi??es de ambiente de laborat?rio a m?dia foi 8,12?0,95 dias. Ao avaliar a capacidade de fixa??o de larvas submetidas a diferentes per?odos de jejum observou-se que as larvas n?o alimentadas mantidas em c?mara climatizada e em condi??es ambientais de laborat?rio foram capazes de se fixar sobre as aves com per?odos de jejum de 6 a 75 dias e 8 a 60 dias, respectivamente. Quando as ninfas foram submetidas a um per?odo de jejum de 15 e 30 dias, nas duas condi??es estudadas, ocorreu o desenvolvimento de ninfas de segundo e terceiro instares. Quando submetidas a 60 dias de jejum verificou-se mortalidade de 28 e 37% em c?mara climatizada e em ambiente de laborat?rio, respectivamente e as sobreviventes n?o se fixaram sobre os hospedeiros. As ninfas de segundo instar submetidas ao jejum de 60 dias desenvolveram, em ambas condi??es estudadas, ninfas de terceiro instar e adultos machos. Ainda no grupo submetido a 60 dias de jejum, as ninfas de terceiro instar desenvolveram adultos (42,42% e 40,54% de machos; 36,36% e 48,65% f?meas nas condi??es de ambiente de laborat?rio e B.O.D., respectivamente) e as demais se desenvolveram em ninfas de quarto instar, que posteriormente, se desenvolveram em adultos f?meas. O n?mero m?dio de ovos produzidos variou entre 46 e 138 ovos nas 18 posturas das f?meas mantidas em ambiente controlado; 41 e 108 ovos nas 9 posturas das f?meas das esta??es chuvosas; e 74 e 138 ovos para as f?meas da esta??o seca, com diferen?a significativa em todas as condi??es experimentais. A dura??o m?dia do ciclo com ocorr?ncia de adultos em N2 foi de 49,05 dias em c?mara climatizada, 53,01 dias nas esta??es secas e 67,41 dias nas esta??es chuvosas. Os resultados obtidos no presente estudo demonstram que os aspectos biol?gicos de A. (P.) miniatus s?o influenciados por fatores clim?ticos, de modo que ocorre um prolongamento do ciclo biol?gico durante a esta??o seca, per?odo que a temperatura ? mais baixa.

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2022-05-05T21:17:13Z No. of bitstreams: 1 2020 - Miguel Angelo Leite Mota J?nior.pdf: 2578202 bytes, checksum: 77a0346aa7dd4ff2aec0a4fcf505e27a (MD5) Made available in DSpace on 2022-05-05T21:17:13Z (GMT). No. of bitstreams: 1 2020 - Miguel Angelo Leite Mota J?nior.pdf: 2578202 bytes, checksum: 77a0346aa7dd4ff2aec0a4fcf505e27a (MD5) Previous issue date: 2020-08-17 CAPES - Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior CNPq - Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico Chicken Infectious Anemia is an acute and contagious disease that leads to immunosuppression in young birds. Being a disease of great impact in the poultry sector, affecting the growth rate, providing a great non-uniformity of the flock, besides the capacity of association of the etiologic agent with other immunosuppressive diseases. The Chicken Infectious Anemia virus (CIAV) has a genetic material of ADN nature, consisting of 3 protein molecules that are crucial components for understanding the behavior of the virus. The objective of this study was to detect the molecular characterization of the CIAV VP1 gene from samples of broilers in industrial and subsistence farms in the state of Rio de Janeiro, Brazil. CIAV was detected using conventional PCR (?Polymerase Chain Reaction?), in different organs, such as: thymus, spleen, liver, cecal tonsil and bursa of Fabricius. From 60 sampled birds (30 from industrial farm and 30 from subsistence) it was possible to detect the presence of CIAV in 21.6% (n = 13/60). In livestock farms, a frequency of 20% (n = 6/30) was observed, while in industrial farms the frequency was 23.33% (n = 7/30) (p> 0.05). The virus was most frequently detected in the thymus and spleen (21.6%, n = 13/60), both in industrial and livestock farms. It is worth mentioning that in none of the evaluated farms the birds were vaccinated against CIAV. Subsequently, two positive samples from the livelihood system and two from the industrial system were sequenced and compared with other sequences available from GenBank to study the genetic diversity of viral strains. Phylogenetic analysis showed that samples 12 and 13 - Subsistence Creation and samples 31 and 36 - Industrial Creation were grouped into a single clade, which reveals that the strains are phylogenetically related. The results presented in this study allow us to conclude that CIAV circulates in farms of industrial creation and subsistence in the state of Rio de Janeiro. However, it cannot be said that livestock livestock are sources of infection for industrial farms. Monitoring using molecular tools for the detection of CIAV in industrial farms is extremely important to assess whether prevention methods are being effective in preventing the introduction of the virus. A Anemia Infecciosa das Galinhas ? uma enfermidade aguda e contagiosa que leva a imunossupress?o em aves jovens. Sendo uma doen?a de grande impacto no setor av?cola afetando a taxa de crescimento proporcionando uma grande desuniformidade do lote, al?m da capacidade de associa??o do agente etiol?gico com outras doen?as imunossupressoras. O v?rus da Anemia Infecciosa das Galinhas (vAIG), apresenta um material gen?tico de natureza DNA, constitu?do por 3 mol?culas proteicas que s?o componentes cruciais para o entendimento do comportamento do v?rus. O objetivo deste estudo foi realizar detec??o a caracteriza??o molecular do gene VP1 do vAIG a partir de amostras de frangos de corte em granjas industriais e de subsist?ncia do estado do Rio de Janeiro, Brasil. O vAIG foi detectado atrav?s da PCR (?Polymerase Chain Reaction?) convencional, em diferentes ?rg?os, como: timo, ba?o, f?gado, tonsila cecal e Bursa de fabr?cius. A partir de 60 aves amostradas (30 de granja industrial e 30 de subsist?ncia) foi poss?vel detectar a presen?a do vAIG em 21,6% (n=13/60). Nas granjas de cria??o de subsist?ncia observou-se uma frequ?ncia de 20% (n=6/30), enquanto nas granjas de cria??o industrial a frequ?ncia foi de 23,33% (n=7/30) (p>0,05). O v?rus foi mais frequentemente detectado no timo e no ba?o (21,6%, n=13/60), tanto em granjas de cria??o industrial como nas de cria??o de subsist?ncia. Vale ressaltar que em nenhuma das granjas avaliadas as aves foram vacinadas contra o vAIG. Subsequentemente, duas amostras positivas do sistema de cria??o de subsist?ncia e duas do sistema industrial foram sequenciadas e comparadas com outras sequ?ncias dispon?veis no GenBank para o estudo da diversidade gen?tica de estirpes virais. A an?lise filogen?tica demonstrou que as amostras 12 e 13 - Cria??o de Subsist?ncia e as amostras 31 e 36 - Cria??o de Industrial foram agrupadas em um ?nico clado, o que revela que as cepas s?o filogeneticamente relacionadas. Os resultados apresentados neste estudo nos permitem concluir que o vAIG circula em granjas de cria??o industrial e de subsist?ncia no estado do Rio de Janeiro. Contudo, n?o se pode afirmar que as aves cria??es de subsist?ncia s?o fontes de infec??o para as granjas industriais. O monitoramento utilizando ferramentas moleculares para detec??o do vAIG em granjas industriais ? de extrema relev?ncia para a avaliar se os m?todos de preven??o est?o sendo eficientes para evitar a introdu??o do v?rus.

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2022-08-02T16:53:02Z No. of bitstreams: 1 2020 - Rodrigo Gredilha Duarte.pdf: 3026424 bytes, checksum: 544bd2418b927996b76cd27d76647586 (MD5) Made available in DSpace on 2022-08-02T16:53:02Z (GMT). No. of bitstreams: 1 2020 - Rodrigo Gredilha Duarte.pdf: 3026424 bytes, checksum: 544bd2418b927996b76cd27d76647586 (MD5) Previous issue date: 2020-06-24 CAPES - Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior The Atlantic Forest is considered one of the 34 biodiversity hotspots on the planet due to the high richness of species and endemism, but the anthropic actions that occur in this biome have contributed to the fauna decline and particularly, infections by hemoparasites can be one more threat to birds of that ecosystem. Thus, understanding the diversity, distribution patterns and evolutionary aspects of these parasites in avian hosts is important for designing actions in conservation programs. In this study, blood samples from 188 wild birds were subjected to genetic analysis with conventional PCR using the 18S rDNA genes to identify Trypanosoma sp, resulting in a total of 10 positive samples; two specimens from the Itatiaia National Park (RJ/MG), Turdus flavipes (?) and T. albicollis, as well as in the capture areas belonging to the Zona da Mata Mineira for six individuals of the species Tachyphonus coronatus, a species of Thamnophilus caerulescens and one from Synallaxis spixi. Five clones from each PCR reaction were submitted to sequencing, totaling 15 sequences from each bird specimen evaluated. All 15 sequences obtained from T. albicollis and T. flavipes were identical to each other. In the six specimens of T. coronatus, five distinct sequences were observed. All sequences of T. caerulescens and S. spixi were identical to each other. Using the strategy of cloning different PCR reactions, it was possible to observe in one of the specimens of T. coronatus two distinct Trypanosoma sequences, which evidences a probable co-infection. The phylogenetic reconstruction showed the grouping of the seven new lineages with parasites of the genus Trypanosoma. The new lineages were grouped into two clades. The lineages JB03, JB04, JB05, ITA01 and ITA02 were grouped together with the species Trypanosoma bennetti. The lines JB01 and JB02 were grouped externally to the clades of the species T. avium, T. cullicavium and T. corvi. The new lineages of trypanosomes found in this study were grouped with lineages recorded in Europe, Asia and Africa. According to the evolutionary distance data obtained in the proposed study, we can observe that the same genetic lineages of Trypanosoma sp. is capable of infecting different species of birds, just as the same species of bird can be infected by more than one lineage of Trypanosoma sp. This study, in an unprecedented way, is the first to access the molecular diversity of parasites of the genus Trypanosoma in birds in Brazil. Until then, all studies related to avian trypanosomes were based mainly on reports of occurrence in blood smears from birds in studies focused on hemoparasites in general. For Brazil, this pioneering study provides possible methods to investigate the diversity of avian trypanosomes and shows the great potential that exists to be explored in this group of parasites. Based on the results of this study, we believe that Brazil, due to its territorial dimensions and the great biodiversity of host birds and insect vectors, may be one of the richest species of avian trypanosomes in the world. A Mata Atl?ntica ? considerada um dos 34 hotspots de biodiversidade do planeta devido ? alta riqueza de esp?cies e endemismo, por?m as a??es antr?picas que ocorrem nesse bioma v?m contribuindo com o decl?nio faun?stico e particularmente, infec??es por hemoparasitos podem ser mais uma amea?a para as aves desse ecossistema. Desta forma, compreender a diversidade, os padr?es de distribui??o e os aspectos evolutivos desses parasitos nos hospedeiros avi?rios ? importante para projetar a??es em programas de conserva??o. Neste estudo, amostras de sangue de 188 aves silvestres foram submetidas ? an?lise gen?tica com PCR convencional por meio dos genes 18S rDNA para identifica??o de Trypanosoma sp, resultando no total de 10 amostras positivas; dois esp?cimes provenientes do Parque Nacional de Itatiaia (RJ/MG), Turdus flavipes (?) e T. albicollis, assim como nas ?reas de captura pertencentes ? Zona da Mata Mineira para seis indiv?duos da esp?cie Tachyphonus coronatus, uma esp?cie de Thamnophilus caerulescens e uma de Synallaxis spixi. Foram submetidos ao sequenciamento 5 clones de cada rea??o de PCR, totalizando 15 sequ?ncias de cada esp?cime de ave avaliada. Todas as 15 sequ?ncias obtidas de T. albicollis e de T. flavipes foram id?nticas entre si. Nos seis esp?cimes de T. coronatus, foram observadas 5 sequ?ncias distintas. Todas as sequ?ncias de T. caerulescens e S. spixi foram id?nticas entre si. Utilizando a estrat?gia de clonagem de rea??es de PCR diferentes foi poss?vel observar em um dos esp?cimes de T. coronatus duas sequ?ncias de Trypanosoma distintas, o que evidencia uma prov?vel coinfec??o. A reconstru??o filogen?tica mostrou o agrupamento das sete novas linhagens com parasitos do g?nero Trypanosoma. As novas linhagens agruparam-se em dois clados. As linhagens JB03, JB04, JB05, ITA01 e ITA02 agruparam-se junto ? esp?cie Trypanosoma bennetti. As linhagens JB01 e JB02 agruparam-se externamente aos clados das esp?cies T. avium, T. cullicavium e T. corvi. As novas linhagens de tripanosomas encontradas neste estudo agruparam-se com linhagens de ocorr?ncia registrada na Europa, ?sia e ?frica. De acordo com os dados de dist?ncia evolutiva obtidos no estudo proposto, podemos observar que uma mesma linhagem gen?tica de Trypanosoma sp. ? capaz de infectar diferentes esp?cies de aves, assim como uma mesma esp?cie de ave pode ser infectada por mais de uma linhagem de Trypanosoma sp. Este estudo, de modo in?dito, ? o primeiro a acessar a diversidade molecular de parasitos do g?nero Trypanosoma em aves no Brasil. At? ent?o, todos os estudos relacionados com a tripanosomas avi?rios basearam-se principalmente em relatos de ocorr?ncia em esfrega?os sangu?neos de aves em estudos voltados aos hemoparasitos em geral. Para o Brasil, este estudo pioneiro fornece m?todos poss?veis para investigar a diversidade de tripanosomas avi?rios e mostra o grande potencial que existe para ser explorado neste grupo de parasitos. Pelos resultados deste estudo, acreditamos que o Brasil, por suas dimens?es territoriais e a grande biodiversidade de aves hospedeiras e insetos vetores, possa ser um dos mais ricos em esp?cies de tripanosomas avi?rios no mundo.

Submitted by Celso Magalhaes (celsomagalhaes@ufrrj.br) on 2022-09-19T13:37:19Z No. of bitstreams: 1 2020 - Ana Paula Martinez de Abreu.pdf: 1765091 bytes, checksum: 6e5b5d16117d027a0f9f2ead2121ddf0 (MD5) Made available in DSpace on 2022-09-19T13:37:20Z (GMT). No. of bitstreams: 1 2020 - Ana Paula Martinez de Abreu.pdf: 1765091 bytes, checksum: 6e5b5d16117d027a0f9f2ead2121ddf0 (MD5) Previous issue date: 2020-03-11 CAPES - Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior This study aimed to perform the molecular characterization of Trypanosoma vivax and Trypanosoma theileri through the 18S and catl genes, as well as to analyze the biochemical profile related to infection for Trypanosoma vivax in cattle from the state of Rio de Janeiro. The samples were collected by means of venocuncture of the coccygeal vein of 389 cattle, and the whole blood collected was placed in tubes containing 10% ethylene-amino-tetracetic acid for molecular analysis and tubes without anticoagulant for biochemistry, being subsequently sent to the Hemoparasites and vectors from the Federal Rural University of Rio de Janeiro where the analyzes were performed. The DNA samples were extracted following the protocol recommended by the manufacturer (Promega?) and subsequently stored in a -20 ? C freezer until the moment of molecular analysis. The serum for biochemical analysis was obtained by centrifugation in a clinical centrifuge at 3,000 rpm for 5 minutes and was later sent to the Clinical Analysis Laboratory of the Experimental Chemotherapy Laboratory in Veterinary Parasitology (LQEPV / UFRRJ), where the serum levels of Glutamyltransferase (GGTT) ), Total Serum Protein (PTS), Albumin (ALB), Total Bilirubin (BT) and Direct (BD), Creatinine (CR), Urea (UR), Alkaline Phosphatase (FAL), Lactate Dehydrogenase (LDH), Aspartate Aminotransferase (AST / TGO) and Alanine Aminotransferase (ALT / TGP) were determined using the automated biochemical analyzer A15 (Biosystems?) and commercial reagent kits according to the equipment used. Molecular analysis was performed using the 18S genes to identify Trypanosoma sp, and catl-like to identify Trypanosoma vivax and T. theileri. After frequency analysis, some samples from each municipality were selected for phylogenetic analysis and comparison of the identity of the sequences of the present study with other sequences found in GenBank. Among the analyzed samples 4.6% (18/389) were positive for Trypanosoma vivax in a blood smear, with no positive samples for Trypanosoma theileri being seen. For the 18S gene, 15.4% (60/389) were positive for Trypanosoma sp, while for the catl gene, 12.8% (50/389) were positive for T. vivax while for T. theileri there was 3.6 % (14/389) of positive animals. Phylogenetic analysis for T. vivax showed 2 clusters defined for the 18S rDNA gene and 4 for the catl gene, whereas for T. theileri, we observed 1 clusters defined for the 18S rDNA gene and 4 for the catl gene, with 15 distinct genotypes. This study aimed to report the first molecular characterization of T. vivax and T. theileri in the state of Rio de Janeiro, as well as the first report of the IB and IF lineage of T. theileri based on the catl gene. In addition, this was also the first report of biochemical analysis of animals naturally infected with Trypanosoma vivax in the state of Rio de Janeiro, where it was possible to verify that LDH was the only significantly altered analysis representing approximately seven times more chance of cattle being positive for T. vivax when this enzyme is increased Este estudo teve como objetivos realizar a caracteriza??o molecular de Trypanosoma vivax e Trypanosoma theileri atrav?s dos genes 18S e catl, bem como analisar o perf?l bioqu?mico relacionado ? infec??o para Trypanosoma vivax em bovinos do estado do Rio de Janeiro. As amostras foram coletadas, por meio de venopun??o da veia cocc?gea de 389 bovinos, sendo o sangue total coletado acondicionado em tubos contendo ?cido etileno-amino-tetrac?tico ? 10% para an?lise molecular e tubos sem anticoagulante para bioqu?mica, sendo posteriormente enviadas ao Laborat?rio de Hemoparasitos e Vetores da Universidade Federal Rural do Rio de Janeiro onde foram realizadas as an?lises. As amostras de DNA foram extra?das seguindo-se protocolo recomendado pelo fabricante (Promega?) e posteriormente armazenadas em freezer -20 ?C at? o momento das an?lises moleculares. O soro para an?lise bioqu?mica foi obtido pela centrifuga??o em centr?fuga cl?nica ? 3.000 rpm por 5 minutos e posteriormente foi enviado ao Laborat?rio de An?lises Cl?nicas do Laborat?rio de Quimioterapia Experimental em Parasitologia Veterin?ria (LQEPV/ UFRRJ), onde os n?veis s?ricos de Gama Glutamiltransferase (GGT), Prote?na S?rica Total (PTS), Albumina (ALB), Bilirrubina Total (BT) e Direta (BD), Creatinina (CR), Ur?ia (UR), Fosfatase Alcalina (FAL), Lactato Desidrogenase (LDH), Aspartato Aminotransferase (AST/TGO) e Alanina Aminotransferase (ALT/TGP) foram determinados usando o analisador bioqu?mico automatizado A15 (Biosystems?) e kits de reagentes comerciais de acordo com o equipamento utilizado. A an?lise molecular foi realizada atrav?s dos genes 18S para identifica??o de Trypanosoma sp, e catl-like (Cathepsina L-like) para identifica??o Trypanosoma vivax e T. theileri. Ap?s an?lise de frequ?ncia, algumas amostras de cada munic?pio foram selecionadas para an?lise filogen?tica e compara??o da identidade das sequ?ncias do presente estudo com outras sequ?ncias encontradas no GenBank. Dentre as amostras analisadas 4,6% (18/389) apresentaram-se positivas para Trypanosoma vivax em esfrega?o sangu?neo, n?o sendo visualizadas amostras positivas para Trypanosoma theileri. Para o gene 18S, 15,4% (60/389) foram positivas para Trypanosomasp, j? para o gene catl, 12,8% (50/389) foram positivos para T. vivax enquanto para T. theileri houve 3,6% (14/389) de animais positivos. A an?lise filogen?tica para T. vivax mostrou 2 clusters definidos para o gene 18S rDNA e 4 para o gene catl, j? para T. theileri, observamos 1clusters definidos para o gene 18S rDNA e 4 para o gene catl, com 15 gen?tipos distintos. O presente estudo teve por finalidade relatar a primeira caracteriza??o molecular de T. vivax e T. theileri no estado do Rio de Janeiro, bem como o primeiro relato da linhagem IB e IF de T. theileri baseado no gene catl. Al?m disso, este tamb?m foi o primeiro relato de an?lise bioqu?mica dos animais infectados naturalmente por Trypanosoma vivax no estado do Rio de Janeiro, onde foi poss?vel verificar que a LDH foi a ?nica an?lise significativamente alterada representando aproximadamente sete vezes mais chance dos bovinos serem positivos para T. vivax quando esta enzima est? aumentada

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2023-01-24T20:12:41Z No. of bitstreams: 1 2020 - Leo Paulis Mendon?a.pdf: 1894334 bytes, checksum: a655cc570a387d0452b2af035dd1fd0b (MD5) Made available in DSpace on 2023-01-24T20:12:41Z (GMT). No. of bitstreams: 1 2020 - Leo Paulis Mendon?a.pdf: 1894334 bytes, checksum: a655cc570a387d0452b2af035dd1fd0b (MD5) Previous issue date: 2020-03-05 CAPES - Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior Ticks are vectors of diseases of great importance for human and veterinary health. The ticks of the group Rhipicephalus sanguineus sense lato (s.l.), known as "common dog tick", are under investigation for their exact taxonomic position, and more recently using molecular techniques it was determined that there are two different lineages, each one addapted to the climate in which they live, temperate or tropical. Monocytic Canine Ehrlichiosis (EMC) is a disease transmitted by ticks Rhipicephalus sanguineus of tropical lineage, caused by the intramonocytic agent Ehrlichia canis and has great importance for the health of dogs and a new zoonotic profile still under investigation, being diagnosed very often in veterinary establishments. The present study deals with an epidemiological investigation, using a real-time PCR technique (qPCR) to detect the agent in question in tick organs samples collected at different altitudes in the State of Rio de Janeiro. Monthly surveys were made to the municipalities of Paracambi, Barra do Pira?, Petr?polis and Teres?polis, visiting residences by non-probabilistic convenience sampling in the urban and rural areas of each municipality and selected for dogs, after being allowed to participate in studies, who answered an epidemiological questionnaire using information about animals and the environment at the time. Ticks were searched for 15 minutes in animals and in the environment for the same ammount of time, put in test tubes capped with cotton, identified, conditioned in B.O.D. chamber and dissected the next day. The organs of each tick were stored in preservative RNAlater? in 1.5 ml microtubes at -20 ? C until the extraction of the genetic material. In all, 412 ticks were collected, among which 86 engorged females of R. sanguineus were dissected. Detection of the agent using qPCR revealed a positivity of 15 out of 86 dissected ticks (17.71%), and among the organs, 15 out of 86 cuts of the intestine and salivary gland were positive (17.71%), while only 10 among 86 ovary samples were positive (11.62%). Variables intrinsic to animals were did not influence in the infection of their ticks by E. canis. Ticks in the dry season were 3.97 times more likely to be infected with E. canis than in the rainy season (CI: 0.89 - 17.71, p

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2022-07-21T17:29:34Z No. of bitstreams: 1 2019 - Michele Bahia do Vale Silva.pdf: 2232740 bytes, checksum: 322f1c2ee1b47d848889c9f5d45a6270 (MD5) Made available in DSpace on 2022-07-21T17:29:34Z (GMT). No. of bitstreams: 1 2019 - Michele Bahia do Vale Silva.pdf: 2232740 bytes, checksum: 322f1c2ee1b47d848889c9f5d45a6270 (MD5) Previous issue date: 2019-10-02 CAPES - Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior This work is divided into two chapters and aims 1) to characterize the importance of rickettsia Anaplasma marginale and the protozoan parasites Babesia bovis and Babesia bigemina in the occurrence of cattle tick fever (CTF) among calves in the Northwest region of Minas Gerais and 2) to evaluate the relationship between A. marginale genetic diversity and the anaplasmosis outbreaks occurrence in dairy farms same region. In the first chapter, Blood samples from 300 calves were collected and after DNA extraction Nested-PCR was performed using oligonucleotide primers that amplified fragments of the genes sbp-4 (B. bovis) and rap-1a (B. bigemina) and Semi-Nested for the gene msp5 (A. marginale). Blood smears were made from these animals. The prevalence of A. marginale 55,66% (167/300), B. bigemina. 15,33% (46/300) and B. bovis 4,0 % (12/300) of calves examined by techinques of PCR. Parasitic forms of A. marginale and B. bigemina were found in 36,33% and 2,66% of the blood smears while B. bovis was not detected. Infection rates by age in group 1 (10-70 days) were 41.12%, 26.16% and 3.73% for A. marginale, B. bigemina and B. bovis, respectively. While in group 2 (> 70-300 days) the prevalences were 63.73%, 9.32% and 4.14% for the same agents, respectively. Fifteen animals with fever, weight loss, jaundice and anemia were examined and nested-PCR confirmed the simple infection by A. marginale. For chapter II, Thirty previously positive calves (msp5 gene) for A. marginale with asymptomatic (n = 24) and acute (n = 6) infection were selected for the study of genetic diversity through semi-nested PCR targeting the msp1? gene. After sequencing of the samples, the genotype and tandem repeats structures were determined. Thirteen different strains were found: : ??FFF (9 animals), 13 27 27 (3 animals), ? 27 18 (3 animals), ??? 195 31 (3 animals), ? 22 13 18 (3 animals), 80 FFFF (2 animals), ? 22 13 13, ????, M????F, 42 25 25 31, QQQM, BQBQBM, 1617FF (1 animal each). Genotype E was predominant in 93.33% of the samples (28/30), followed by genotypes C (3.33%) and G (3.33%). A new repeated tandem structure was described in the present work named BRA 1 (TDSSSASGVLSQSGQASTSSQLG). The ??FFF strain was present in six febrile animals that showed clinical sintoms of anaplasmosis like weight loss, jaundice and anemia. In the bovine herd studied anaplasmosis is the main cause of CTF. High genetic diversity of A. marginale occurred in persistently infected animals. However, during outbreaks of bovine anaplasmosis, low genetic diversity was observed. The ??FFF strain in northwestern Minas Gerais was associated with acute anaplasmosis. Genotype E was predominate. Este trabalho est? dividido em dois cap?tulos e tem por objetivos: 1) Caracterizar a import?ncia da rickettsia Anaplasma marginale e dos protozo?rios Babesia bovis e Babesia bigemina na ocorr?ncia do complexo da Tristeza Parasit?ria Bovina (TPB) em bezerros na regi?o Noroeste de Minas Gerais 2) avaliar a rela??o entre a diversidade gen?tica de A. marginale e a ocorr?ncia de surtos de anaplasmoses em bezerros da mesma regi?o. No primeiro cap?tulo foram coletadas 300 amostras sangu?neas de bezerros e ap?s extra??o de DNA foi realizada Nested- PCR utilizando oligonucleot?deos iniciadores que amplificam fragmentos dos genes sbp-4 (B. bovis) e rap-1a (B. bigemina) e a Semi-Nested para o gene msp5 (A. marginale). Esfrega?os sangu?neos foram confeccionados destes animais. A preval?ncia de A. marginale foi de 55,66% (167/300), B. bigemina 15,33% (46/300) e B. bovis 4,0% (12/300) nos animais examinados por t?cnicas de PCR. Formas parasit?rias de A. marginale e B. bigemina foram encontradas em 36,33% e 2,66% dos esfrega?os sangu?neos, enquanto B. bovis n?o foi detectado. As taxas de infec??es por faixa et?ria no grupo 1 (10-70 dias) de bezerros foram de 41,12%, 26,16% e 3,73% para A. marginale, B. bigemina e B. bovis, respectivamente. Enquanto no grupo 2 (> 70-300) as preval?ncias foram de 63,73%, 9,32% e 4,14% para os mesmos agentes, respectivamente. Foram examinados 15 animais com febre, perda de peso, mucosas hipocoradas, icter?cia e anemia, sendo confirmados posteriormente por nested-PCR a infec??o simples por A. marginale, todos animais pertenciam ao grupo 2. Para o segundo cap?tulo foram selecionados 30 bezerros previamente positivos (gene msp5) para A. marginale com infec??o assintom?tica (n=24) e aguda (n=6) para o estudo da diversidade gen?tica atrav?s da semi-nested PCR com alvo no gene msp1?. Ap?s o sequenciamento das amostras foi determinado o gen?tipo e estruturas em tandem repeats. Treze diferentes estirpes foram encontradas ??FFF (9 animais), 13 27 27(3 animais), ? 27 18 (3 animais), ??? BRA1 31 (3 animais), ? 22 13 18 (3 animais), 80 FFFF (2 animais), ? 22 13 13, ????, M????F, 42 25 25 31, QQQM, BQBQBM, 1617FF (um animal cada). O gen?tipo E foi predominante em 93,33% das amostras (28/30), seguidos pelos os gen?tipos C (3,33%) e G (3,33%). Uma nova estrutura repetida em tandem foi descrita no presente trabalho nomeada BRA 1 (TDSSSASGVLSQSGQASTSSQLG). A estirpe ??FFF esteve presente em seis animais com sinais cl?nicos de anaplasmose aguda com sinais cl?nicos de febre, perda de peso, icter?cia e anemia. No Rebanho bovino estudado a anaplasmose ? a principal causa de Tristeza Parasit?ria bovina. Alta diversidade gen?tica de A. marginale ocorreu em animais persistentemente infectados. Entretanto durante surtos de anaplasmose bovina foi observada baixa diversidade gen?tica. A estirpe ??FFF na regi?o noroeste de Minas Gerais foi associada a anaplasmose aguda. O gen?tipo E foi predomina-te.

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2021-11-23T19:12:54Z No. of bitstreams: 1 2019 - Tha?s Alves Fernandes.pdf: 1086460 bytes, checksum: cadad6fd66db182f3da30b96025cdb59 (MD5) Made available in DSpace on 2021-11-23T19:12:54Z (GMT). No. of bitstreams: 1 2019 - Tha?s Alves Fernandes.pdf: 1086460 bytes, checksum: cadad6fd66db182f3da30b96025cdb59 (MD5) Previous issue date: 2019-05-08 CAPES - Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior The intentional or incidental frauds in meat and meat products by the substitution of species of animals of butcher, occur in a recurring form, constituting as a pressing concern of governmental entities focused on the guarantee of the quality of food and consumer rights. This practice can cause economic losses as well as high negative impacts for the food producing industry and for consumers, whether for moral, ethical, religious or public health issues. The development of new technologies allowed industrial establishments to create more elaborate, diversified products with increasingly complex characteristics. Several analytical methods has been developed with the intention of detecting fraud by replacing types of meat from one animal species with another of smaller value, helping to recognize the authenticity of meat products. The aim of the present work was to standardize a Real Time Polymerase Chain Reaction (qPCR) Multiplex and High Resolution Melting (HRM) technique to detect and differentiate meat types from butchers and derivatives. The singleplex reaction of the specie Gallus gallus showed limit of detection of ten copies, with a coefficient of determination (R?) of 0.991 and an efficiency of 105%. The reaction of the species Sus scrofa presented detection limit of 1 copie, with R? 0.999 and an efficiency of 105%. Reactions of the species Equus caballus and Bos indicus showed limit of detection of 1 copy, R? of 0,999 and 0,996 and efficiency of 98 and 94,8%, respectively. The results showed that the singleplex reactions were specific, sensitive and applicable in the identification of animal species in meat and meat products. The multiplex reaction can be used to detect to identify different species of butcher animals in a single reaction. As fraudes intencionais ou incidentais em carnes e produtos c?rneos pela substitui??o de esp?cies de animais de a?ougue, ocorrem de forma recorrente, constituindo-se como uma preocupa??o premente de entidades governamentais voltadas para a garantia da qualidade de alimentos e direitos do consumidor. Esta pr?tica pode causar preju?zos econ?micos al?m de elevado impacto negativo para a ind?stria produtora de alimentos e para os consumidores, seja por quest?es, morais, ?ticas, religiosas ou relacionadas ? sa?de p?blica. O desenvolvimento de novas tecnologias permitiu aos estabelecimentos industriais a cria??o de produtos mais elaborados, diversificados e com caracter?sticas cada vez mais complexas. Diversos m?todos anal?ticos v?m sendo desenvolvidos, com a inten??o de detectar fraudes pela substitui??o de tipos de carne de uma esp?cie animal por outra de menor valor auxiliando no reconhecimento da autenticidade dos produtos c?rneos. O presente trabalho teve como objetivo padronizar uma Rea??o em Cadeia da Polimerase em Tempo Real (qPCR) Multiplex e ?High Resolution Melting? (HRM) para detectar e diferenciar tipos de carnes de animais de a?ougue e derivados. A rea??o singleplex da esp?cie Gallus gallus apresentou limite de detec??o dez c?pias do genoma, com um coeficiente de determina??o (R?) de 0,991 e uma efici?ncia de 105%. A rea??o da esp?cie Sus scrofa apresentou um limite de detec??o de 1 c?pia, com R? 0,999 e efici?ncia de 105%. As rea??es das esp?cies Equus caballus e Bos indicus apresentaram limite de detec??o de uma c?pia do genoma, R? de 0,999 e 0,996 e efici?ncia de 98% e 94,8%, respectivamente. Os resultados obtidos demonstraram que as rea??es singleplex, foram espec?ficas, sens?veis e aplic?veis na identifica??o de esp?cies animais em produtos c?rneos. A rea??o multiplex pode ser utilizada para detec??o e identifica??o de diferentes esp?cies de animais de a?ougue em uma ?nica rea??o.

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2021-08-31T00:26:11Z No. of bitstreams: 1 2018 - Renata Vit?ria Campos Costa.pdf: 5276179 bytes, checksum: 37eaa035f7cf5629b0fd719954616b3e (MD5) Made available in DSpace on 2021-08-31T00:26:11Z (GMT). No. of bitstreams: 1 2018 - Renata Vit?ria Campos Costa.pdf: 5276179 bytes, checksum: 37eaa035f7cf5629b0fd719954616b3e (MD5) Previous issue date: 2018-12-06 Trypanosomiasis is a disease caused by species of the genus Trypanosoma. In cattle trypanosomiasis is caused by Trypanosoma vivax that originates in tropical Africa, where the disease represents an obstacle to the development of livestock. The spread of disease to the Americas occurred during European colonization in the 1830s, and the displacement of the herds and the adaptation of the parasite to mechanical transmission by hematophagous flies allowed the spread of this parasite. In Brazil, T. vivax infection in cattle was reported for the first time in the State of Par?, in 1944, and the presence of the parasite has been reported in several states. This study aimed to report the epidemiological and clinical-pathological aspects of T. vivax natural infection in cattle in the state of Rio de Janeiro. Sixteen foci of Trypanosoma vivax infection were diagnosed in cattle in 13 municipalities in the state of Rio de Janeiro between April 2016 and October 2018. Of these, 12 outbreaks were studied based on epidemiological data, clinical-pathological signs and laboratory tests. Clinically affected animals presented anemia, decreased appetite, significant decrease in milk production, progressive weight loss, weakness, reproductive abnormalities, diarrhea and neurological signs. In some animals the picture evolved to death. The laboratory diagnosis (parasitological, molecular, necropsy and histopathological examinations) was performed on 284 whole blood samples and tissues collected from 280 cattle. Parasitological exams performed on leukocyte - layer smears were positive in 25 samples from six of the 12 foci studied. Molecular diagnosis using conventional polymerase chain reaction (cPCR), carried out on whole blood samples, detected 46 positive samples in five of the 12 foci and in seven other positive samples in tissue fragments collected. The cPCR presented high diagnostic sensitivity when compared to parasitological methods. Of the 12 cows that died, nine presented neurological signs and, on histological examination, inflammatory alterations were found mainly in the central nervous system and heart, characterized by meningoencephalitis and multifocal myocarditis; in the central nervous system of the animals these lesions occurred with greater intensity in the brain in relation to the marrow. Trypanosomiasis in bovine herds in the state of Rio de Janeiro is of concern because of its potential to cause economic losses and, due to the lack of specificity of clinical signs, laboratory tests are fundamental to elucidate the diagnosis. As tripanossom?ases s?o enfermidades causadas por esp?cies do g?nero Trypanosoma. Em bovinos a tripanossom?ase ? causada pelo Trypanosoma vivax que tem origem na ?frica tropical, onde a enfermidade representa um obst?culo para o desenvolvimento da pecu?ria. A expans?o da enfermidade para as Am?ricas ocorreu durante a coloniza??o europeia, na d?cada de 1830, e o deslocamento dos rebanhos e a adapta??o do parasito ? transmiss?o mec?nica por moscas hemat?fagas possibilitou a dissemina??o dessa parasitose. No Brasil a infec??o por T. vivax em bovinos foi relatada pela primeira vez no estado do Par?, em 1944, e atualmente a presen?a do parasito j? foi registrada em diversos estados. Este estudo teve como objetivo relatar os aspectos epidemiol?gicos e cl?nico-patol?gicos da infec??o natural por T. vivax em bovinos no estado do Rio de Janeiro. Foram diagnosticados 16 focos de infec??o Trypanosoma vivax em bovinos, em 13 munic?pios do estado do Rio de Janeiro, entre abril de 2016 a outubro de 2018. Destes, 12 focos foram estudados com base nos dados epidemiol?gicos, sinais cl?nico-patol?gicos e exames laboratoriais. Os animais clinicamente afetados apresentaram anemia, diminui??o do apetite, diminui??o significativa na produ??o de leite, emagrecimento progressivo, fraqueza, altera??es reprodutivas, diarreia e sinais neurol?gicos. Em alguns animais o quadro evoluiu para a morte. O diagn?stico laboratorial (exames parasitol?gico, molecular, necropsia e histopatol?gico) foi realizado em 284 amostras de sangue total e tecidos coletados de 280 bovinos. Os exames parasitol?gicos realizados em esfrega?os de capa leucocit?ria resultaram positivos em 25 amostras provenientes de seis dos 12 focos estudados. O diagn?stico molecular atrav?s da rea??o em cadeia de polimerase convencional (cPCR), realizado em amostras de sangue total, detectou 46 amostras positivas em cinco dos 12 focos e em mais sete amostras positivas em fragmentos de tecidos coletados. A cPCR apresentou elevada sensibilidade diagn?stica quando comparada aos m?todos parasitol?gicos. Das 12 vacas que morreram, nove apresentaram sinais neurol?gicos e, ao exame histol?gico, foram encontradas altera??es inflamat?rias principalmente no sistema nervoso central e cora??o, caracterizadas por meningoencefalite e miocardite linfohistiocit?ria multifocal; no sistema nervoso central dos animais estas les?es ocorreram com maior intensidade no c?rebro em rela??o ? medula. A tripanossom?ase em rebanhos bovinos no estado do Rio de Janeiro traz preocupa??o devido ao seu potencial em causar preju?zos econ?micos e, em face da inespecificidade dos sinais cl?nicos, os exames laboratoriais s?o fundamentais para elucidar o diagn?stico.

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2022-05-05T20:25:38Z No. of bitstreams: 1 2018 - Gabriela Lopes Vivas Vitari.pdf: 3853711 bytes, checksum: 045cc88cc08fab46bb8b4148ce9329b3 (MD5) Made available in DSpace on 2022-05-05T20:25:38Z (GMT). No. of bitstreams: 1 2018 - Gabriela Lopes Vivas Vitari.pdf: 3853711 bytes, checksum: 045cc88cc08fab46bb8b4148ce9329b3 (MD5) Previous issue date: 2018-03-28 CNPq - Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico Equine piroplasmosis, caused by the hemoparasite Theileria equi, is endemic in Brazil and is considered one of the major equine diseases in the world. The objective of the present study was to investigate the genetic diversity of geographically distinct isolates of 18S rRNA gene sequences of T. equi in naturally infected horses from the five Brazilian regions, characterize the hypervariable regions of the T. equi 18S rRNA sequences from this study and to elucidate the taxonomic relationships between T. equi isolates from Brazil and of the world. Equine blood samples were collected in the five regions of the country. The genomic DNA was extracted and the samples were submitted to real-time PCR for the selection of positive samples. Subsequently, the selected samples were submitted to a conventional PCR for amplification of the 18S rRNA gene (~1600 bp) of T equi, the products being sequenced. The sequences were aligned using ClustalW, and the alignment was submitted to the Maximum Likelihood reconstruction method with molecular distances estimated by the General Time Reversible Model, using 1000 replicates. From this alignment a similarity matrix was developed and the hypervariable regions were observed. Phylogenetic analyzes were performed in three different ways: using T. equi sequences from Brazil, using sequences from different species of Theileria and Babesia, and using T. equi sequences from all over the world. Twenty-three partial sequences of the 18S rRNA gene of T. equi were obtained, of which thirteen were unique variants of the gene in Brazil. The hypervariable gene regions V2, V4 and V8 were identified in the study samples, with V4 being the most variable. These hypervariable regions were confirmed by entropy analysis. Two distinct genotypes circulating in the Brazilian regions were identified. Only a single genotype was observed in the North region, while in the other regions both were observed. Samples were grouped into two clades (A and C), most of the study sequences being clustered in clade C. In another tree, all T. equi sequences grouped into group 1, different from the other theilerias and babesias, present in groups 2 and 3. In relation to the T. equi samples of the world, three clades were observed: A, B and C. This study adds important information on the genetic diversity of T. equi in horses from all regions of Brazil based on the 18S rRNA sequence. A piroplasmose equina, causada pelo hemoparasito Theileria equi, ? end?mica no Brasil e ? considerada uma das principais doen?as de equinos no mundo. O objetivo do presente estudo foi investigar a diversidade gen?tica de isolados geograficamente distintos de sequ?ncias do gene 18S rRNA de T. equi em equinos naturalmente infectados provenientes das cinco regi?es do Brasil, caracterizar as regi?es hipervari?veis das sequ?ncias 18S rRNA de T. equi deste estudo e elucidar as rela??es taxon?micas entre os isolados de T. equi do Brasil e do mundo. As amostras de sangue dos equinos foram coletadas nas cinco regi?es do pa?s. O DNA gen?mico foi extra?do e as amostras foram submetidas a PCR em tempo real para a sele??o das amostras positivas. Posteriormente, as amostras selecionadas foram submetidas a PCR convencional para a amplifica??o do gene 18S rRNA (~1600 pb) de T. equi, sendo os produtos sequenciados. As sequ?ncias foram alinhadas utilizando o ClustalW, e o alinhamento foi submetido ao m?todo de reconstru??o da M?xima Verossimilhan?a com dist?ncias moleculares estimadas pelo General Time Reversible Model, utilizando 1000 repeti??es. Desse alinhamento foi desenvolvida uma matriz de similaridade e foram observadas as regi?es hipervari?veis. As an?lises filogen?ticas foram realizadas de tr?s formas distintas: utilizando sequ?ncias de T. equi do Brasil, utilizando sequ?ncias de diferentes esp?cies de Theileria e Babesia, e utilizando sequ?ncias de T. equi provenientes de todo o mundo. Foram obtidas vinte e tr?s sequ?ncias parciais do gene 18S rRNA de T. equi, sendo treze delas variantes ?nicas do gene no Brasil. As regi?es do gene hipervari?veis V2, V4 e V8 foram identificadas nas amostras do estudo, sendo a V4 a mais vari?vel. Essas regi?es hipervari?veis foram confirmadas pela an?lise de entropia. Foram identificados dois gen?tipos distintos circulantes nas regi?es do Brasil. Apenas na regi?o Norte foi observado um ?nico gen?tipo, enquanto que nas demais regi?es observaram-se ambos. As amostras se agruparam em dois clados (A e C), estando a maioria das sequ?ncias do estudo agrupadas no clado C. Em outra ?rvore, todas as sequ?ncias de T. equi se agrupam dentro do grupo 1, diferente das demais theilerias e babesias, presentes nos grupos 2 e 3. Em rela??o ?s amostras de T. equi do mundo, tr?s clados foram observados: A, B e C. Este estudo acrescenta importantes informa??es sobre a diversidade g?nica de T. equi em equinos de todas as regi?es do Brasil baseado na sequ?ncia 18S rRNA.

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2021-08-30T23:53:33Z No. of bitstreams: 1 2018 - Renata Lins da Costa.pdf: 2024444 bytes, checksum: dbd9cb0e5d394366e1c63faf2ee951d9 (MD5) Made available in DSpace on 2021-08-30T23:53:33Z (GMT). No. of bitstreams: 1 2018 - Renata Lins da Costa.pdf: 2024444 bytes, checksum: dbd9cb0e5d394366e1c63faf2ee951d9 (MD5) Previous issue date: 2018-02-28 Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico (CNPq) Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) Funda??o Carlos Chagas Filho de Amparo ? Pesquisa do Estado do Rio de Janeiro (FAPERJ) The aim of the present study was to characterize molecularly Ehrlichia canis using the membrane glycoprotein genes involved in the host parasite interaction (gp19, gp36, and p28), and also, the thermo unstable elongation factor (tuf) gene for characterization together with the 16S rDNA gene from dog blood samples, as well as to evaluate the distribution of E. canis in dogs from the mesoregions of Rio de Janeiro state. 267 blood samples from dogs from the state of Rio de Janeiro were obtained and tested by the Real-Time PCR (qPCR) targeting a fragment of 93 base pairs (bp) of 16S rDNA gene as a screening method for detection of the positive samples. Before evaluating the genetic diversity of E. canis, the samples were submitted to the molecular detection by 16S rDNA-qPCR. A total of 267 blood samples of dogs from the state of Rio de Janeiro were collected. Among the samples analyzed, 42.3% (n=113/267) were qPCR positive for 16S rDNA gene of E. canis. The average value of Cq observed in positive samples was 34,1 ? 5,1, ranging between 18 to 40 cycles. The detection limit of the qPCR was 10 copies of the plasmid per ?L containing a 16S rDNA gene from E. canis. The determination coefficient of the seven dilutions tested in the standard curve was 99.9% and efficiency de 95.7%. When performing gp19-PCR and gp36-PCR, 100% (n=113/113) and 5.3% (n=6/113) of the samples were positive, respectively. The six PCR positive samples for the 16S rDNA gene also amplified the p28 gene. Only one PCR positive sample for the three genes (gp19, gp36 and p28) in each of the six mesoregions were selected and subjected to analysis of amino acid and nucleotide sequences. The frequency in each mesoregion for the 113 animals positive for E. canis, by the 16S rDNA gene, was 59,29% (n=67/113) belonged to Metropolitan mesoregion, 13.27% (n=15/113) to South Fluminense, 15.04% (n=17/113) to Northern Fluminense, 5.3% (n=6/113) Center Fluminense, 4.42% (n=5/113) to Northwest Fluminense and 2.65% (n=3/113) to Coastal Baixada. The characterization based on the gp19 gene demonstrated that this gene is highly conserved and the samples showed 100% similarity with the Brazilian and worldwide isolates available in GenBank. Using the gp36 gene, it was possible to observe that there are polymorphic points between the sequences. Cluster analysis shows the formation of 3 clades, the sequences of the present study were allocated to the genogroup of the United States, suggesting a genetic similarity between the Brazilian and North American strains. In the protein analysis, all the samples presented repeated sequences (Tandem Repeat Sequences), with 11 replicates. Seven high entropy amino acid sites were observed, with variations of Hx: 0.5 to 0.67 generating mean entropy of 0.6 detected along the TRP36, suggesting a high degree of polymorphism. The ratio of non-silent to silent mutations using both genes showed significant genetic differences (p 0.05) with E. canis positivity. The investigation of gene differences in the isolates of E. canis becomes useful for understanding the parasite-host relationship; in addition, determining the distribution of this bacterium in the evaluated municipalities is advantageous for the updating of the epidemiological chain of canine ehrlichiosis. Os objetivos do presente estudo foram de caracterizar molecularmente Ehrlichia canis, utilizando os genes de glicoprote?nas de membrana (gp19, gp36, p28) e o gene Fator de Alongamento termo inst?vel (tuf) para caracteriza??o g?nica juntamente com o gene 16S rDNA das amostras de sangue de c?es naturalmente infectados, bem como avaliar a distribui??o de E. canis e fatores associados em c?es oriundos das mesorregi?es do estado do Rio de Janeiro. Foram testadas 267 amostras de sangue de c?es do estado do Rio de Janeiro, pela rea??o em cadeia da polimerase em tempo real (qPCR) com alvo em um fragmento de 93 pares de base (pb) do gene 16S rDNA para a detec??o de E. canis. Para realiza??o da caracteriza??o molecular, as amostras positivas nesta primeira an?lise foram testadas por PCRs convencionais para os genes gp19 (414 pb), gp36 (814 pb) e p28 (840 pb). Uma amostra de cada mesorregi?o amplificada para os genes gp19, gp36 e p28 foram purificadas e sequenciadas, utilizando-se o m?todo Sanger. Antes de avaliar a diversidade gen?tica de E. canis, as amostras foram submetidas ? detec??o molecular por 16S rDNA-qPCR. Foram coletadas 267 amostras de sangue de c?es do estado do Rio de Janeiro. Entre as amostras analisadas, 42,3% (n = 113/267) foram positivas para qPCR para o gene 16S rDNA de E. canis. O valor m?dio de Cq observado nas amostras positivas foi de 34,1 ? 5,1, variando entre 18 e 40 ciclos. O limite de detec??o do qPCR foi de 10 c?pias do plasm?deo por ?L contendo um gene 16S rDNA de E. canis. O coeficiente de determina??o das sete dilui??es testadas na curva padr?o foi de 99,9% e efici?ncia de 95,7%. Ao realizar PCR para os genes gp19 e gp36, 100% (n=113/113) e 5,3% (n=6/113) das amostras foram positivas, respectivamente. As seis amostras positivas para PCR para o gene 16S rDNA tamb?m amplificaram o gene p28. Apenas uma amostra PCR positiva para os tr?s genes (gp19, gp36 e p28) em cada uma das seis mesorregi?es foi selecionada e submetida ? an?lise de sequ?ncias de amino?cidos e nucleot?deos. A freq??ncia em cada mesorregi?o para os 113 animais positivos para E. canis, pelo gene 16S rDNA, foi de 59,29% (n=67/113) para mesorregi?o Metropolitana, 13,27% (n =15/113) para regi?o Sul Fluminense, 15,04% (n=17/113) para mesorregi?o Norte Fluminense, 5,3% (n= 6/113) para mesorregi?o Centro Fluminense, 4,42% (n=5/113) na mesorregi?o Noroeste Fluminense e 2,65% (n= 3/113) na Baixada Litor?nea. A caracteriza??o baseada no gene gp19 demonstrou que este gene ? altamente conservado e as amostras apresentaram 100% de similaridade com as sequ?ncias brasileiros e mundiais dispon?veis no GenBank. Utilizando o gene gp36, foi poss?vel observar que h? pontos polim?rficos entre as sequ?ncias. Na an?lise de agrupamentos nota-se a forma??o de 3 clados, as sequ?ncias do presente estudo alocaram-se no genogrupo dos Estados Unidos, sugerindo similaridade gen?tica entre as sequ?ncias brasileiras e norte-americanas. Na an?lise de prote?nas, todas as amostras apresentaram sequ?ncias repetidas (?Tandem Repeat Sequence?), com 11 repeti??es. Foram observados sete s?tios de amino?cidos de alta entropia, com varia??es de Hx: 0,5 a 0,67 gerando uma m?dia de entropia de 0,6 detectados ao longo de TRP36, o que sugere um alto grau de polimorfismo. A raz?o das muta??es n?o-silenciosas sobre as silenciosas, utilizando ambos genes mostrou diferen?as gen?ticas significativas (p0,05) com a positividade de Ehrlichia canis nas sequ?ncias analisadas. A investiga??o de diferen?as g?nicas nos isolados de E. canis torna-se ?til para o entendimento e conhecimento da rela??o parasito-hospedeiro; al?m disso determinar a distribui??o desta bact?ria nos munic?pios avaliados torna-se vantajoso para a atualiza??o da cadeia epidemiol?gica da erliquiose canina.

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2021-07-27T22:07:47Z No. of bitstreams: 1 2018 - Nath?lia Alves de Senne.pdf: 1372014 bytes, checksum: b3a0d29f4c00d1a4f8dcaf631f17b3a0 (MD5) Made available in DSpace on 2021-07-27T22:07:47Z (GMT). No. of bitstreams: 1 2018 - Nath?lia Alves de Senne.pdf: 1372014 bytes, checksum: b3a0d29f4c00d1a4f8dcaf631f17b3a0 (MD5) Previous issue date: 2018-02-27 Leishmaniasis represents a group of zoonotic, infectious diseases that put at risk more than 350 million people worldwide. Its transmission occurs during the blood feeding of females of sand flies infected by the etiological agent of the disease, the protozoan of the genus Leishmania. Due to the fundamental role played by the vector in the chain of transmission of the disease, the objective of this study was to evaluate the ecology and epidemiology of sandflies present in endemic areas for Leishmaniasis in the municipality of Serop?dica. Sand flies were captured through a CDC-type light trap, located in peridomiciliary areas, for three consecutive days during a one-year period in five city neighborhoods, located in rural and peri-urban areas, where cases of Leishmaniasis detection in humans. Phlebotomines were identified for the species after the clarification and diaphanization process, according to Galati (2003). The indexes of temperature, relative air humidity and precipitation index were collected monthly on the site of the meteorological institute (INMET) and correlated with the number of specimens collected. An epidemiological questionnaire was applied to an owner of each residence where a trap was placed to obtain data about his knowledge about leishmaniasis and evaluation of environmental conditions and factors that contribute to the greater proliferation of sandflies. The main ecological data evaluated were of richness and abundance, using the coefficients of Shannon and Simpson; for statistical correlation, the Mann-Whitney and Kruskal-Wallis tests were used. A total of 17371 specimens were collected, identified in five species: Lutzomyia intermedia, Lutzomyia migonei, Lutzomyia pessoai, Lutzomyia whitmani and Lutzomyia fischeri. Of the specimens collected 57.5% (9984) were males and 42.5% (7387) were females. Val?o das Lou?as was the site that presented the largest number of specimens collected (9096). The highest Shannon index was recorded in Campo Lindo (H = 0.7021) and the most abundant species was L. intermedia (69.1%). A significant difference was observed in relation to the number of sandflies collected in rural and periurban areas (p

Submitted by Leticia Schettini (leticia@ufrrj.br) on 2021-08-16T15:17:11Z No. of bitstreams: 1 2018 - Patr?cia Gonzaga Paulino.pdf: 1868248 bytes, checksum: dc2e4d01c72edc3a37722a8afe55ab6b (MD5) Made available in DSpace on 2021-08-16T15:17:12Z (GMT). No. of bitstreams: 1 2018 - Patr?cia Gonzaga Paulino.pdf: 1868248 bytes, checksum: dc2e4d01c72edc3a37722a8afe55ab6b (MD5) Previous issue date: 2018-02-21 Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) Funda??o de Apoio ? Pesquisa do Estado do Rio de Janeiro (FAPERJ) Canine vector diseases are frequently found in brazilian veterinary clinics. This group represents a varied complex of illness including anaplasmosis, ehrlichiosis, babesiosis, rangeliosis, hepatozoonosis among other diseases. Many of these pathogens cause severe clinical conditions, death of patients and in some cases, have zoonotic potential. There are a variety of diagnostic methods for these microorganism on the market. Molecular techniques are based on the detection of DNA from the etiological agents directly from the blood with high sensitivity and specificity, allowing the detection even under conditions of very low parasitemias, which are often characteristic of asymptomatic carriers. Also allows the distinction between different species of parasites. In this study, two molecular methods were developed for the detection of Babesia vogeli in blood samples from dogs from the Itaguai Microregion, based on quantitative real-time polymerase chain reaction (qPCR) technology and polymerase chain reaction (cPCR) with a specific target in hsp70 gene from B. vogeli. After the optimization and standardization of the techniques, it was observed that the qPCR developed in this study was efficient and presented high precision and sensitivity in the detection of B. vogeli. Comparative studies were performed between the 18S rDNA and hsp70 targets using cPCR, where it was possible to observe that there are no statistically significant differences between the two targets, but in the sequencing a false positive for B. vogeli was detected in the 18S rDNA sequence, a proximal hemoprotozoal related to Rangelia vitalli. The cPCR and qPCR techniques were compared for the detection of B. vogeli and it was observed that qPCR has higher sensitivity. In addition, an epidemiological study was carried out, which showed three factors associated with B. vogeli infection among these were young animals (less than 5 years old), the infestation by ticks and the absence of shelters. As doen?as vetoriais que acometem c?es s?o diagnosticadas com alta frequ?ncia nas cl?nicas veterin?rias brasileiras. Esse grupo representa um complexo variado de mol?stias incluindo anaplasmose, ehrliquiose, babesiose, rangeliose, hepatozoonose entre outras enfermidades. Muitos desses pat?genos causam severas condi??es cl?nicas, ?bito de pacientes e ainda, em alguns casos, possuem potencial zoon?tico. Existe uma variedade de m?todos diagn?sticos para esses agentes no mercado. As t?cnicas moleculares t?m como base a detec??o do DNA dos agentes etiol?gicos diretamente do sangue com alta sensibilidade e especificidade, possibilitando a detec??o mesmo em condi??es de parasitemias muito baixas, que s?o muitas vezes caracter?sticas de animais portadores assintom?ticos, e permite a distin??o entre diferentes esp?cies de parasitos. Neste estudo foi desenvolvido dois m?todos moleculares para detec??o de Babesia vogeli em amostras de sangue de c?es provenientes da Microrregi?o de Itagua?, baseados na tecnologia de rea??o em cadeia da polimerase quantitativa em tempo real (qPCR) e a rea??o em cadeia da polimerase convencional (cPCR) com alvo espec?fico no gene hsp70 de B. vogeli. Ap?s os processos de otimiza??o e padroniza??o das t?cnicas, foi observado que a qPCR desenvolvida neste estudo foi eficiente, apresentando alta precis?o e sensibilidade na detec??o de B. vogeli. Foi realizado estudos comparativos entre os alvos 18S rDNA e hsp70 usando cPCR, onde, foi poss?vel observar que n?o existem diferen?as estat?sticas significativas entre os dois alvos. Por?m no sequenciamento foi detectado um falso positivo para B. vogeli na sequ?ncia 18S rDNA, um hemoprotozo?rio proximamente relacionado com Rangelia vitalli na regi?o. As t?cnicas de cPCR e qPCR foram comparadas quanto a detec??o de B. vogeli e foi observado que a qPCR tem maior sensibilidade. Al?m disso, foi realizado em paralelo um estudo epidemiol?gico, o qual evidenciou tr?s fatores associados a infec??o presen?a de B. vogeli sendo estes animais jovens (com menos de 5 anos), infesta??o por carrapatos e a aus?ncia de abrigos.

Submitted by Celso Magalhaes (celsomagalhaes@ufrrj.br) on 2021-05-18T13:58:48Z No. of bitstreams: 1 2017 - Roberta Caroline Alvarenga Tolomelli.pdf: 1156139 bytes, checksum: f473634631348eb53ff79188b874b6be (MD5) Made available in DSpace on 2021-05-18T13:58:48Z (GMT). No. of bitstreams: 1 2017 - Roberta Caroline Alvarenga Tolomelli.pdf: 1156139 bytes, checksum: f473634631348eb53ff79188b874b6be (MD5) Previous issue date: 2017-08-16 Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES Canine babesiosis and ehrlichiosis are transmitted by the brown dog tick, Rhipicephalus sanguineus sensu lato (s.l.). These hemoparasites are well distributed in Brazil and the canine ehrlichiosis is more frequent than babesiosis. The aim of this study was to assess the frequency of Ehrlichia canis and Babesia vogeli in unfed ticks from two counties of Itagua? microregion, Rio de Janeiro. 369 R. sanguineus s.l. ticks were analyzed in peridomiciliary environment, including adults (194), nymphs (92) and larvae (83). The DNA samples were extracted through a HotSHOT modified method. The molecular detection of E. canis was made by a PCR that amplifies a 409 bp fragment of Disulfide oxidoreductase (dsb) gene of Ehrlichia spp. For B. vogeli detection, the samples were tested by a PCR that amplifies a 523 bp fragment of heat shock protein 70kDa (hsp70) gene. We obtained 41 (11.11%) positive samples for E. canis, 11 (5,67%) adults, 8 (8,69%) nymphs ans 22 (26,5%) larvae (p

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2019-03-19T17:37:03Z No. of bitstreams: 1 2017 - D?bora Martins dos Santos.pdf: 1544900 bytes, checksum: 2029812eaa2eb53116da77515d19516e (MD5) Made available in DSpace on 2019-03-19T17:37:05Z (GMT). No. of bitstreams: 1 2017 - D?bora Martins dos Santos.pdf: 1544900 bytes, checksum: 2029812eaa2eb53116da77515d19516e (MD5) Previous issue date: 2017-03-07 The avian infectious bronchitis (AIB) is a disease caused by a single-stranded RNA virus pertaining to the coronaviridae family of the Gammacoronavirus genera. AIB is a disease of great economic significance to aviculture worldwide, not only because of the injury it causes to the breeders, but also because it is a differential diagnosis for several important pathologies that may affect poultry. The goals of the present research were to develop a RT-PCR RFLP technique capable of safely detecting and discriminating the strains of the IBV of the Massachusetts and BR serotypes from the trachea, kidney and intestines tissues in poultry farms of broilers and layers in the estate of Rio de Janeiro. In this study, 117 samples of intestines, kidneys and/or tracheae were collected from a total of 50 birds originating from 4 farms (three broiler farms and one layer farm) in the estate of Rio de Janeiro. The total RNA was extracted from all the samples and subjected to a reverse transcription process for the cDNA synthesis. The cDNA samples were subjected to the RT-qPCR technique. It was found that 92% of the birds (n= 46/50) were positive in at least one of the organs for the chicken bronchitis virus (vBIG). The RT-PCR RFLP technique developed in this work was able to detect a frequency of 94,64% (n= 53/56) of the samples belonging to the BR-I serotype, 3,7% (n= 2/56) of the samples showed fragments with patterns similar to what is expected from the Massachusetts Serotype and 1,78% (n= 1/56) showed patterns from both serotypes (Massachusetts and BR). There was no significant difference in the frequency of molecular detection of vBIG in the tissues of the trachea, kidney and intestines of the broiler and layer poultries from farms in the estate of Rio de Janeiro. The RT-PCR RFLP technique was able to safely detect and distinguish the Massachusetts and BR serotypes. The high frequency of the BR serotype demonstrates that, although vaccinated, the birds developed the disease, indicating that a new vaccine protocol must be addressed, since the majority of the strains found belong to a genetically distinct serotype. A Bronquite Infecciosa das Galinhas (BIG) ? uma doen?a causada por um v?rus RNA de fita simples pertencente ? Fam?lia Coronavidae, do g?nero Gammacoronavirus. A BIG ? uma doen?a de alta import?ncia econ?mica para a avicultura mundial, n?o s? por causar preju?zos aos plant?is acometidos pela doen?a, como tamb?m, por ser diagn?stico diferencial de diversas patologias importantes na sanidade av?cola. O objetivo da presente pesquisa foi desenvolver uma t?cnica de RT-PCR RFLP capaz de detectar e discriminar com seguran?a as cepas do vBIG do sorotipo Massachusetts e BR a partir de tecidos da traqueia, rim e intestino em granjas de frango de corte e postura, no Estado do Rio de Janeiro. Neste estudo, foram coletadas 117 amostras de intestino, rim e/ou traqueia de um total de 50 aves provenientes de quatro granjas (tr?s granjas de corte e uma granja de postura), no estado do Rio de Janeiro. O RNA total foi extra?do de todas as amostras e submetidos a uma rea??o de transcri??o reversa, para realizar a s?ntese do cDNA. As amostras de cDNA foram submetidas ? t?cnica de RT-qPCR. Desta maneira, constatou-se que 92% das aves (n = 46/50) foram positivas em pelo menos um dos ?rg?os para o v?rus da bronquite das galinhas (vBIG). A t?cnica de RT-PCR RFLP desenvolvido por esse trabalho foi capaz de detectar uma frequ?ncia de 94,64% (n = 53/56) das amostras pertencentes ao sorotipo BR-I, 3,57% (n = 2/56) das amostras apresentaram fragmentos com padr?o semelhante ao esperado do sorotipo Massachusetts e 1,78% (1/56) apresentou padr?o para ambos os sorotipos (Massachusetts e BR). N?o houve diferen?a significativa entre a frequ?ncia da detec??o molecular do vBIG nos tecidos da traqueia, rim e intestino de frangos de corte e galinhas de postura de granjas no estado do Rio de Janeiro. A t?cnica de RT-PCR RFLP foi capaz de detectar e difere.nciar com seguran?a os sorotipos Massachusetts e BR. A alta frequ?ncia do sorotipo BR demonstra que apesar de vacinadas, as aves desenvolveram a doen?a, sugerindo, que um novo protocolo vacinal deva ser abordado, j? que a maioria das cepas encontradas pertence a um sorotipo geneticamente distinto.

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2018-03-27T18:48:03Z No. of bitstreams: 1 2017 - Tays Araujo Camilo.pdf: 3555895 bytes, checksum: 8b6818596f8654186472296bc3b528aa (MD5) Made available in DSpace on 2018-03-27T18:48:03Z (GMT). No. of bitstreams: 1 2017 - Tays Araujo Camilo.pdf: 3555895 bytes, checksum: 8b6818596f8654186472296bc3b528aa (MD5) Previous issue date: 2017-03-07 CNPq Infectious Bronchitis Virus (vBIG) is a highly contagious viral agent among the species Gallus gallus, being considered one of the pathologies that most affects small and lare world poultry producers. Vaccination as a form of prevention to this disease, has not been effective, due to the great genetic variability found. As in the world, in Brazil there are already several studies characterizing the genetic variability found in this virus. In Rio de Janeiro, poultry farming suffers from the disease, but there is no recent study to analyze the genetic variability of the strains found, as a way of supporting epidemiological studies that may help in the future, in the choice of Protocols. The objective of this study was to characterize the S1 gene of the Chicken Infectious Bronchitis virus (vBIG) in chickens farms in the southern region of Rio de Janeiro, as well as to evaluate the genetic variability of the S1 gene in a BIG outbreak on a farm Of laying hens of the northern region of. Tissue samples were collected from animals presenting BIG-compatible clinical signs, such as weight loss, respiratory problems and low productivity. Samples of collected tissues were stored in solution of RNA Later and formalin 10% buffered for molecular and histopathological diagnoses, respectively. In order to perform the molecular analyzes, RNA extraction techniques, as well as RT-PCR (Reverse Transcriptase, Polymerase Chain Reaction), real-time PCR (RT-qPCR), Nested PCR (RT-nPCR), and Sequencing of some samples collected. For the histopathological analyzes, cleavage and preparation of microscopy slides were performed. The results obtained from the sequencing were compared to the sequence of the Ma5 vaccine strain as reference strain, in addition to other sequences deposited on GenBank, exhibiting a variable identity percentage of 72.41% to 100%. Among the samples from each farm, there was variability, even within the same bird. All sequences analyzed were described as genotype 1, belonging to strains 1 and 11; In the comparison of amino acids, there were changes in the hypervariable regions of the virus in the sequences classified as BR-1, being able to explain the low cross-protection that have been occurring in Brazilian plants. These results demonstrated the importance of virus identification, since it becomes an important tool in epidemiological surveillance and in the elaboration of efficient vaccine protocols. O v?rus da Bronquite Infecciosa das Galinhas (vBIG) ? um agente viral altamente contagioso entre a esp?cie Gallus gallus, sendo considerado uma das patologias que mais acomete a avicultura mundial. A vacina??o como forma de preven??o a esta doen?a, n?o tem sido eficaz, devido a grande variabilidade gen?tica encontrada. Assim como no mundo, no Brasil tamb?m j? existem diversos estudos caracterizando a variabilidade gen?tica encontrada neste v?rus. No Rio de Janeiro, a cria??o av?cola sofre com a doen?a, por?m ainda n?o existe um estudo recente com o objetivo de analisar a variabilidade gen?tica das cepas encontradas, como uma forma de apoio a estudos epidemiol?gicos, que possam auxiliar no futuro, na escolha de protocolos vacinais mais adequados. Este estudo teve como objetivo realizar a caracteriza??o molecular do gene S1 do v?rus da Bronquite Infecciosa das Galinhas (vBIG) em granjas de frangos de corte na regi?o Sul fluminense, como tamb?m avaliar a variabilidade gen?tica do gene S1 em um surto de BIG em uma granja de poedeiras da regi?o Norte fluminense. Foram realizadas coletas de tecidos dos animais que apresentavam sinais cl?nicos compat?veis com BIG, como perda de peso, problemas respirat?rios e baixa produtividade. Amostras de tecidos coletados foram armazenadas em solu??o de RNA Later e formalina 10% tamponada para os diagn?sticos molecular e histopatol?gico, respectivamente. Para realiza??o das an?lises moleculares foram utilizadas t?cnicas de extra??o de RNA, bem como RT-PCR (?Reverse Transcriptase, Polymerase Chain Reaction?), PCR em tempo real (RT-qPCR), ?Nested? PCR (RT-nPCR), e sequenciamento de algumas amostras coletadas. Para as an?lises histopatol?gicas foram realizadas clivagem e elabora??o de l?minas de microscopia. Os resultados obtidos do sequenciamento foram comparados a sequ?ncia da cepa vacinal Ma5, como cepa refer?ncia, al?m de outras sequencias depositadas no GenBank, apresentando um percentual de identidade vari?vel de 72,41% a 100%. Dentre as amostras de cada granja, houve variabilidade, at? mesmo dentro de uma mesma ave. Todas as sequencias analisadas foram descritas como gen?tipo 1, pertencentes as linhagens 1 e 11; Na compara??o de amino?cidos, houve mudan?as nas regi?es hipervari?veis do v?rus nas sequ?ncias classificadas como BR-1, podendo explicar a baixa prote??o-cruzada que v?m ocorrendo nos plant?is brasileiro. Estes resultados demonstraram a import?ncia da identifica??o g?nica do v?rus, pois se torna uma ferramenta importante na vigil?ncia epidemiol?gica e em elabora??es de protocolos vacinais que sejam eficientes.

Submitted by Celso Magalhaes (celsomagalhaes@ufrrj.br) on 2019-02-26T17:39:51Z No. of bitstreams: 1 2017 - N?dia Valesca Biral de Oliveira Peixoto de Mello.pdf: 2451067 bytes, checksum: 8baa15bc902ba124aab56edccdd0abde (MD5) Made available in DSpace on 2019-02-26T17:39:51Z (GMT). No. of bitstreams: 1 2017 - N?dia Valesca Biral de Oliveira Peixoto de Mello.pdf: 2451067 bytes, checksum: 8baa15bc902ba124aab56edccdd0abde (MD5) Previous issue date: 2017-03-03 Leishmaniasis in equidae is a reality in the world, and this family has been described as reservoir for the disease since 1927. These animals develop a cutaneous form, of a benign nature, most of them spontaneous cure. In the Federal District (FD), traction equids are widely used as means of transportation and work, being inserted directly inside the city, in close contact with dogs and humans. They live mostly in community pens, extremely unhealthy environments, with few hygienic-sanitary conditions, which attract large numbers of insects. The present study aimed to determine the prevalence of anti-Leishmania sp. Through the techniques of Indirect Immunofluorescence (IFI) and the Immunoenzymatic Assay (ELISA) in traction equids in the FD and to describe possible epidemiological variables associated with this prevalence. The FD equidae population is estimated to be 1682 animals (SEAGRI / FD), distributed in 20 administrative regions (AR). The sample size was calculated according to Medronho (2009) and the animals were randomly selected in each AR, of which 178 (43.31%) were seroreagents in the ELISA, 197 (47.93%) in the IFI and 111 (27.01%) reagents in both ELISA /IFI techniques. A semi-structured questionnaire was used and the variables associated with the prevalence of antibodies were analyzed by Qui-square in the BioEstat program, version 5.0. The following variables were considered significant (p 0.05): sex, length of stay with the owner and whether the animals had already left the FD, when the animals were analyzed in the IFI; Sex, locality, methods of reducing insects in the environment, residence time with the owner and if the animals have already left the FD, when analyzing the reactive animals in the ELISA and; Sex, to use repellent in the animals, if the animals have already left the FD, ocular mucosa and, entered in areas with stream, when the reagents in the IFI / ELISA were analyzed. New studies that may help to better understand the participation of equidae in the maintenance of the epidemiological cycle of Leishmania sp will be of great importance and will allow the recommendation of more efficient control measures. The present work collaborated in demonstrating the possibility of equidae being infected and presenting an important role in the epidemiological chain of Leishmania sp and in the maintenance of the disease in the territory of the Federal District. A leishmaniose em equ?deos ? uma realidade no mundo, sendo esta fam?lia descrita como reservat?rio para a doen?a desde 1927. Esses animais desenvolvem uma forma cut?nea, de car?ter benigno, tendo em sua maioria a cura espont?nea. No Distrito Federal (DF), equ?deos de tra??o s?o amplamente usados como meio de transporte e trabalho, estando inseridos diretamente dentro da cidade, em contato ?ntimo com c?es e seres humanos. Vivem na maioria das vezes em currais comunit?rios, ambientes extremamente insalubres, com poucas condi??es higi?nico-sanit?rias, que atraem grande quantidade de insetos. O presente trabalho teve como objetivo determinar a preval?ncia de anticorpos anti-Leishmania sp. atrav?s das t?cnicas de Imunofluoresc?ncia Indireta (RIFI) e do Ensaio Imunoenzim?tico (ELISA) em equ?deos de tra??o no DF e descrever poss?veis vari?veis epidemiol?gicas associadas com esta preval?ncia. A popula??o de equ?deos do DF ? estimada em 1682 animais (SEAGRI/DF), distribu?dos em 20 regi?es administrativas (RA). O c?lculo do tamanho da amostra foi realizado de acordo com Medronho (2009) e os animais foram selecionados de forma aleat?ria em cada RA, sendo 178 (43,31%) sororreagentes no ELISA, 197 (47,93%) na RIFI e 111 (27,01%) reagentes em ambas a t?cnicas ELISA/RIFI. Foi utilizado um question?rio semiestruturado e as vari?veis associadas com a preval?ncia de anticorpos foram analisadas pelo Qui quadrado, no programa BioEstat, vers?o 5.0. As seguintes vari?veis foram consideradas significativas (p 0,05): sexo, tempo de perman?ncia com o propriet?rio e se os animais j? deixaram o DF, quando analisados os animais reagentes na RIFI; sexo, localidade, m?todos de redu??o de insetos no ambiente, tempo de perman?ncia com o propriet?rio e se os animais j? deixaram o DF, quando analisados os animais reagentes no ELISA e; sexo, usar repelente nos animais, se os animais j? deixaram o DF, mucosa ocular e, entrada em ?reas com c?rrego, quando analisados os reagentes na RIFI/ELISA. Novos estudos que possam auxiliar no melhor entendimento da participa??o dos equ?deos na manuten??o do ciclo epidemiol?gico de Leishmania sp ser?o de grande import?ncia e possibilitar?o a preconiza??o de medidas de controle mais eficientes. O presente trabalho colaborou ao demonstrar a possibilidade de equ?deos estarem se infectando e apresentando um papel importante na cadeia epidemiol?gica de Leishmania sp e na manuten??o da doen?a no territ?rio do DF.

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2019-01-31T18:26:42Z No. of bitstreams: 1 2017 - Maristela Peckle Peixoto.pdf: 10701979 bytes, checksum: faffb982c158894c0c385ae44c6a8b7c (MD5) Made available in DSpace on 2019-01-31T18:26:42Z (GMT). No. of bitstreams: 1 2017 - Maristela Peckle Peixoto.pdf: 10701979 bytes, checksum: faffb982c158894c0c385ae44c6a8b7c (MD5) Previous issue date: 2017-02-22 CAPES - Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior CNPq - Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico FAPERJ - Funda??o Carlos Chagas Filho de Amparo ? Pesquisa do Estado do Rio de Janeiro This study aimed to perform the molecular characterization of the B. caballi rap-1 gene and to analyze the degree of heterogeneity of the T. equi 18S rDNA gene of naturally infected horses in the state of Rio de Janeiro. Complete sequences of the T. equi 18S rDNA gene (n = 20) were obtained. The PCR products were cloned and sequenced. Phylogenetic analyzes were performed on ARB-Silva basis. For the molecular characterization of a fragment of the B. caballi rap-1 gene, 32 cELISA and qPCR positive equine samples were used. A nested PCR was developed to amplify a fragment of the rap-1 gene and a conventional PCR, aiming the amplification of the complete gene. The products were sequenced. Sequences were submitted to the Maximum-Likelihood reconstruction method with Poisson model and 1000 replicates. In the analysis of T. equi, twelve different isolates were observed in equines from the state of Rio de Janeiro and were grouped into two of the three clades formed (I, II), most of them grouped in clade I (70%, n = 14/20). The sequences of T. equi are grouped within a clade different from other Theilerids. It was possible to amplify a fragment of the B. caballi rap-1 gene from 23 sequences using nested PCR, identified as single sequences. One of the sequences showed twelve mutation points. In cluster analysis, the Brazilian samples belonged to clade A, being analyzed as amino acids (152a) or nucleotides (1244 bp). The aim of the second chapter was to evaluate the dinamic of infection of T. equi in R. (B.) microplus fed in chronically infected horses. The larvae obtained from a hemoparasite-free colony were used in the infestation of an equine chronically infected by T. equi. Ticks and the horse blood were collected every two days. Ticks were dissected and stored in pools according to the pos-infestation day, evolutionary stage and gender. Some of these specimens were stored entire. Salivary gland and ovary samples were stored properly for Transmission Electron Microscopy. Some stages, prior to ecdysis, were stored in B.O.D. to molt. Samples were tested by qPCR. Regarding to entire tick samples, more than 90% of the samples were positive for T. equi. The mean of parasite load in entire ticks and salivary glands presented gradual increasing with the progress of the feeding period. Of the 45 salivary gland samples, 77.78% were positive for T. equi and had a higher infection rate at the end of the experimental period. It was found freshly molted ticks positives for T. equi by qPCR, indicating transestadial transmission. 84,38% of R. (B.) microplus ovary samples were considered positives for T. equi by molecular tools. Males of R. (B.) microplus were found parasiting the equine for 60 days. This fact associated with the high positivity for T. equi indicates males as an epidemiologically important gender. Transmission micrographs revealed morphologic structures compatible with a Piroplasmid in the acini of salivary glands and ovaries. The phylogenetic analysis of the T. equi 18S rDNA gene and of B. caballi rap-1 gene showed that different isolates co-exist in the same area in Rio de Janeiro, Brazil. Experimently, it was possible to observe that R. (B.) microplus could be infected with T. equi and that the forms of development of this parasite are capable of multiply inside salivary glands and, possibly, ovaries. Os objetivos deste trabalho foram caracterizar molecularmente Theileria equi e Babesia caballi de amostras de equinos do Rio de Janeiro, bem como realizar um estudo da din?mica da infec??o de T. equi em Rhipicephalus (Boophilus) microplus. No primeiro cap?tulo realizou-se a caracteriza??o molecular de um fragmento do gene rap-1 de B. caballi e a an?lise do grau de heterogeneidade do gene 18S rDNA em T. equi em amostras de equinos naturalmente infectados, do estado do Rio de janeiro. Foram obtidas vinte sequ?ncias do gene 18S rDNA de T. equi. As an?lises filogen?ticas foram realizadas baseadas no ARB-Silva. Para a caracteriza??o molecular de um fragmento do gene rap-1 de B. caballi foram utilizadas amostras positivas por cELISA e qPCR. Uma nested PCR foi desenvolvida para amplificar um fragmento do gene rap-1, e uma PCR convencional, para o gene completo. As sequ?ncias de amino?cidos e nucleot?deos sequenciados foram submetidas ao m?todo Maximum-Likelihood. Na an?lise das sequ?ncias do 18S rDNA de T. equi, doze diferentes isolados foram observados em equinos no estado do Rio de Janeiro, sendo agrupados em dois dos tr?s clados formados (I, II). As sequ?ncias de T. equi se agrupam dentro de um clado diferente dos demais Theiler?deos. Uma das 23 sequ?ncias de amino?cidos do gene rap-1 apresentou doze pontos de muta??o. Na an?lise de agrupamentos, as amostras brasileiras agruparam no clado A, sendo estas analisadas como amino?cidos (152 aas) ou nucleot?deos (1244 pb). No segundo cap?tulo foram avaliados aspectos da din?mica da infec??o de T. equi em R. (B.) microplus alimentados em equino cronicamente infectado. Larvas de uma col?nia livre de hemoparasitos foram utilizadas. Carrapatos e sangue do equino foram coletados de dois em dois dias. Os carrapatos foram dissecados e armazenados inteiros em ?pools? de acordo com o dia p?s-infesta??o, est?gio evolutivo e g?nero. Amostras de gl?ndula salivar e ov?rio foram armazenados para Microscopia Eletr?nica de Transmiss?o. As amostras foram testadas pela qPCR. Com rela??o aos carrapatos inteiros, mais de 90% das amostras foram positivas para T. equi. A carga parasit?ria m?dia das amostras de carrapatos inteiros e gl?ndulas salivares apresentaram aumento gradual com o avan?ar do per?odo de alimenta??o. Das 45 amostras de gl?ndula salivar, 77,78% foram positivas para T. equi e tiveram maior taxa de infec??o no final do per?odo experimental. Foi poss?vel observar positividade em carrapatos rec?m emergidos, ninfas e adultos, indicando a ocorr?ncia de transmiss?o transestadial. A positividade de amostras de ov?rios foi de 84,38%. A maior perman?ncia de machos sobre o equino, associado ? alta positividade deste tipo de amostra indicam que o macho pode ter participa??o significativa na transmiss?o. Visualizaram-se estruturas morfologicamente compat?veis com um piroplasma atrav?s de micrografia de transmiss?o em ov?rios e gl?ndulas salivares de em R. (B.) microplus. A an?lise do gene 18S rDNA de T. equi e do gene rap-1 de B. caballi de amostras brasileiras demonstrou que diferentes isolados coexistem na mesma ?rea. Experimentalmente, observou-se que R. (B.) microplus ? capaz de se infectar com T. equi e que as formas multiplicam-se nas gl?ndulas salivares e, possivelmente, em ov?rios.

Submitted by Celso Magalhaes (celsomagalhaes@ufrrj.br) on 2017-10-19T13:49:16Z No. of bitstreams: 1 2016 - Claudia Bezerra da Silva.pdf: 8032175 bytes, checksum: ef71dd2a0e7801e9000e116c822a3a00 (MD5) Made available in DSpace on 2017-10-19T13:49:16Z (GMT). No. of bitstreams: 1 2016 - Claudia Bezerra da Silva.pdf: 8032175 bytes, checksum: ef71dd2a0e7801e9000e116c822a3a00 (MD5) Previous issue date: 2016-03-11 CAPES - Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior and investigate the circulation of this agent in dogs in the Itaguai microregion, Rio de Janeiro, Brazil, and dogs and ticks in two provinces of the island of Cuba, analyzing epidemiological aspects associated with infections caused by this bacterium in dogs. A new real-time polymerase chain reaction method (qPCR) was patterned to target the citrate synthase gene (gltA) for the identification of A. platys in naturally infected dogs. The primers and probe were designed to amplify a fragment of 84 base pairs based on gltA gene sequences of A. platys available in GenBank. 186 blood samples of dogs from Itaguai microregion, Rio de Janeiro, Brazil, were tested by qPCR. The same samples were tested by cytology and nested polymerase chain reaction (nPCR, 16S rDNA) to determine the performance of qPCR front of these techniques. 17.20% of the samples tested positive by qPCR were significantly more than that detected by nPCR (13.98%). The qPCR technique was more specific than cytology, due to false-positive results obtained by optical microscopy. The prevalence of A. platys in dogs from Itaguai microregion was 14.4%. Dogs less than six months, infested by ticks, that spend the most of the time restrict to domestic environment and without shelter are factors associated with infection by this hemoparasite in dogs in the study area. During research, A. platys held in Cuba, 100 blood samples were collected from residents dogs in four cities located in the provinces of Havana and Mayabeque. When inspecting the animals, found ticks were collected, identified and carefully grouped, forming a total of 49 pools. DNA extracted from blood samples from dogs and ticks were subjected nPCR (16S rDNA). Positive samples in nPCR were also subjected to conventional PCR (gltA gene), and the products were sequenced. Only the species Rhipicephalus sanguineus sensu lato was found in Cuban dogs and 10.2% (n=5/49) of these ticks added to 16.0% (n=16/100) dogs were considered positive for A. platys. All sequences analyzed of the gltA and 16S rDNA genes, respectively, showed a 99-100% identity with sequences from A. platys reported in other countries. Phylogenetic analysis showed two clusters defined for the 16S rDNA gene and three clusters defined for the gltA gene. Based on the gltA gene, the deduced amino acid sequence showed two points of non-synonymous mutations at positions 88 and 168 compared to the reference sequence DQ525687. A preliminary study on the epidemiological aspects associated with infection with A. platys showed no statistical association with the variables studied (p> 0.05). This study also to report the first evidence of A. platys in both dogs and ticks in Cuba also presents for the first time the development of a new qPCR method that contributes to the advancement of research involving A. platys. The epidemiological study in Brazil allowed us to identify significant factors in the occurrence of canine anaplasmosis, while in Cuba, it can be concluded that more research is needed to assess what the deciding factors in the transmission and spread of A. platys in that country. platys, e investigar a circula??o deste agente em c?es na microrregi?o de Itagua?, Rio de Janeiro, Brasil, e c?es e carrapatos em duas prov?ncias da ilha de Cuba, analisando aspectos epidemiol?gicos associados ? infec??o causada por esta bact?ria em c?es. Um novo m?todo de rea??o em cadeia da polimerase em tempo real (qPCR) foi padronizado com alvo no gene citrato sintase (gltA) para a identifica??o de A. platys em c?es naturalmente infectados. Os oligoiniciadores e a sonda foram desenhados para amplificar um fragmento de 84 pares de base baseado em sequ?ncias do gene gltA de A. platys dispon?veis no GenBank. 186 amostras de sangue de c?es da microrregi?o de Itagua?, Rio de Janeiro, Brasil, foram testados pela qPCR. As mesmas amostras foram testadas pela citologia e rea??o em cadeia da polimerase nested (nPCR, 16S rDNA) para determinar o desempenho da qPCR frente ? essas t?cnicas. 17,20% das amostras testadas pela qPCR foram positivas, significativamente mais do que detectado pela nPCR (13,98%). A t?cnica de qPCR foi mais espec?fica que a citologia, em virtude dos resultados falsopositivos obtidos pela microscopia ?ptica. A preval?ncia de A. platys em c?es da microrregi?o de Itagua? foi de 14,4%. C?es com menos de seis meses, infestados por carrapatos, que possam maior tempo restrito ao ambiente dom?stico e sem abrigo s?o fatores associados a infec??o por este hemoparasito em c?es na regi?o do estudo. Durante investiga??o de A. platys realizada em Cuba, 100 amostras de sangue foram coletadas de c?es residentes em quatro cidades localizadas nas prov?ncias de Habana e Mayabeque. Ao inspecionar os animais, carrapatos encontrados foram coletados, identificados e criteriosamente agrupados, formando um total de 49 pools. Amostras de DNA extra?das do sangue dos c?es e de carrapatos foram submetidas a nPCR (16S rDNA). Amostras positivas na nPCR foram tamb?m submetidas a PCR convencional (gene gltA), e os produtos foram sequenciados. Somente a esp?cie Rhipicephalus sanguineus sensu lato foi encontrada em c?es cubanos, e 10,2% (n=5/49) desses carrapatos somado aos 16,0% (n=16/100) de c?es foram considerados positivos para A. platys. Todas as sequ?ncias analisadas dos genes gltA e 16S rDNA, respectivamente, mostraram uma identidade de 99-100% com sequ?ncias de A. platys reportadas em outros pa?ses. A an?lise filogen?tica mostrou dois clusters definidos para o gene 16S rDNA e tr?s clusters definidos para o gene gltA. Com base no gene gltA, a sequ?ncia de amino?cidos deduzidos demonstrou dois pontos de muta??es n?o-sin?nimas nas posi??es 88 e 168 comparados com sequ?ncia de refer?ncia DQ525687. Um estudo preliminar sobre os aspectos epidemiol?gicos associados com a infec??o por A. platys demonstrou nenhuma associa??o estat?stica com as vari?veis avaliadas (p > 0,05). O presente estudo al?m de relatar a primeira evid?ncia de A. platys em ambos c?es e carrapatos em Cuba, tamb?m apresenta pela primeira vez o desenvolvimento de um novo m?todo de qPCR que contribui para o avan?o da pesquisa envolvendo A. platys. O estudo epidemiol?gico realizado no Brasil permitiu identificar fatores importantes na ocorr?ncia da anaplasmose canina, enquanto em Cuba, pode-se concluir que mais investiga??es s?o necess?rias para avaliar quais os fatores decisivos na transmiss?o e dispers?o de A. platys nesse pa?s.

Submitted by Celso Magalhaes (celsomagalhaes@ufrrj.br) on 2017-10-03T16:25:38Z No. of bitstreams: 1 2016 - Viviane Figueira Marques.pdf: 1883187 bytes, checksum: 9996d5ea41ecd4f93424c808c4d82ba6 (MD5) Made available in DSpace on 2017-10-03T16:25:38Z (GMT). No. of bitstreams: 1 2016 - Viviane Figueira Marques.pdf: 1883187 bytes, checksum: 9996d5ea41ecd4f93424c808c4d82ba6 (MD5) Previous issue date: 2016-03-02 CAPES - Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior Staphylococcus spp. plays an important role in the etiology of bovine mastitis. Staphylococcus aureus is considered the most relevant specie due to the production of virulence factors such as ?slime?, which is required for biofilm formation. This study aimed to detect the phenotypic expression of the biofilm formation in 20 S. aureus isolates from bovine mastitis, to detect and quantify the expression of genes involved in its production and regulation, as well as to detect the phenogenotypic resistance to beta-lactam in order to evaluate the possible relation between biofilm production and antimicrobial resistance. The isolates were characterized by MALDI-TOF and phenogenotypic identification assays. Also they were submitted to the phenotypic tests to evaluate biofilm production and the susceptibility to beta-lactams. Minimum Inhibitory Concentration (MIC), Minimum Bactericidal Concentration (MBC) and Minimum Inhibitory Concentration in the Biofilm (MICB) were determined to three isolates presenting distinct biofilm production. Futherly, a PCR for the detection of ?slime? production genes (icaA and icaD), Bap protein (bap), beta-lactamase (blaZ) and protein altered penicillin-binding (mecA). Also, the Agr system was typified (agr I, agr II, agr III and agr IV) and its regulator (agr RNAIII) was detected. Scanning Electron Microscopy (SEM) was performed to determine the most suitable time interval for biofilm analysis. Real-time PCR (qPCR) was performed at the chosen times to quantify the expression of icaA, icaD and hld genes in the three studied isolates. All 20 isolates were biofilm producers and most presented icaA and icaD genes. Only one isolate presented the bap gene. The agr gene type II presented a prevalence of 70%. The SEM showed gradual changes in bacterial arrangement during the biofilm formation along the phases of the growth curve. The peak was reached at the stationary phase. Transcriptional analysis revealed increased expression of ica genes at 8 h of growth and of hld at 24 h. However, for the N-365 strain the ica expression was of low yield. For this study, the penicillin resistance was related to the production of beta-lactamase otherwise the high MBC detected for cefoxitin may be associated to biofilm protection, evidentiated by the fact that the isolates have MICB values higher than MICs tested for planktonic cells Staphylococcus spp. tem papel importante na etiologia da mastite bovina. Staphylococcus aureus ? considerada a esp?cie mais relevante devido ? produ??o de fatores de virul?ncia, tais como ?slime?, o que favorece a forma??o do biofilme. O presente trabalho teve por objetivo detectar a express?o fenot?pica da forma??o de biofilme em 20 isolados de S. aureus oriundos de mastite bovina, detectar e quantificar a express?o dos genes envolvidos na sua produ??o e regula??o, al?m de detectar a resist?ncia fenogenot?pica aos beta-lact?micos para avalia??o da poss?vel rela??o da produ??o de biofilme com a resist?ncia antimicrobiana. Os isolados foram caracterizados atrav?s de testes fenogenot?picos e MALDI-TOF, submetidos ?s provas fenot?picas de detec??o da forma??o de biofilme e avalia??o da suscetibilidade aos beta-lact?micos. A Concentra??o Inibit?ria M?nima (CIM), Concentra??o Bactericida M?nima (CBM) e a Concentra??o Inibit?ria M?nima no Biofilme (CIMB) foram determinadas para tr?s isolados selecionados com base na varia??o da intensidade da produ??o de biofilme. Posteriormente, todos os isolados foram submetidos ? t?cnica de PCR para detec??o dos genes de produ??o de ?slime? (icaA e icaD), prote?na Bap (bap), beta-lactamase (blaZ) e prote?na ligante de penicilina alterada (mecA). Al?m de detec??o do sistema regulador Agr (agr RNAIII) e da tipifica??o do sistema Agr (agr I, agr II, agr III e agr IV). Foi realizada Microscopia Eletr?nica de Varredura (MEV) para determinar o intervalo de tempo mais adequado para a an?lise do biofilme. A PCR em tempo real (qPCR) foi realizada nos tempos selecionados para quantificar a express?o dos genes icaA, icaD e hld em tr?s isolados com produ??o variada de biofilme. Todos os isolados foram produtores de biofilme e a maioria apresentou os genes icaA e icaD. Apenas um isolado apresentou o gene bap. O gene agr tipo II mostrou preval?ncia de 70%. A MEV mostrou mudan?as graduais no arranjo bacteriano durante a forma??o de biofilme ao longo das fases da curva de crescimento que atingiu seu pico de forma??o na fase estacion?ria. A an?lise transcricional evidenciou maior express?o dos genes ica no tempo de 8 h de crescimento e hld em 24 h. Contudo, a cepa N-365 mostrou baixa produ??o dos genes ica. Para este estudo, a resist?ncia ? penicilina foi relacionada com a produ??o de beta-lactamase, enquanto a elevada CBM detectada para cefoxitina pode estar associada ? prote??o que o biofilme oferece, epis?dio evidenciado pelo fato dos isolados apresentarem valores de CIMB superiores aos CIMs testados para as c?lulas planct?nicas.

Submitted by Leticia Schettini (leticia@ufrrj.br) on 2016-09-20T14:14:37Z No. of bitstreams: 1 2016 - Naiara de Miranda Bento Rodrigues.pdf: 1508318 bytes, checksum: bbf2aebb5d43033339bf559d73461919 (MD5) Made available in DSpace on 2016-09-20T14:14:37Z (GMT). No. of bitstreams: 1 2016 - Naiara de Miranda Bento Rodrigues.pdf: 1508318 bytes, checksum: bbf2aebb5d43033339bf559d73461919 (MD5) Previous issue date: 2016-02-26 Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico - CNPQ Mastitis adversely affects milk production and in general cows do not regain their full production levels post recovery, leading to considerable economic losses. Moreover the percentage decrease in milk production depends on the specific pathogen that caused the infection and enterobacteria are responsible for this greater reduction. These microorganisms are preferentially found in the habitat of animals in places contaminated with feces, urine, clay and also organic beds. Phenotypic tests are among the currently available methods used worldwide to identify enterobacteria; however they tend to misdiagnose the species despite the multiple tests carried out and they can delay the antibiotic therapy by clinic veterinary. On the other hand the MALDI-TOF MS technique has been attracting attention for its precise identification of several microorganisms at species level. In the current study, 183 enterobacteria were detected in milk (n=47) and fecal samples (n=94) collected from cows; also water (n=23) and milk line samples (n=19) collected from a farm in Rio de Janeiro with the purpose to present the MALDI-TOF MS technique as efficient methodology and also as a ?gold standard? to better understand the possible current biochemical errors in enterobacteria identification considering isolates from bovine environments. This proteomic technique confirmed 92.9% (170/183) of the enterobacteria species identified by biochemical tests that showed high sensitivity (> 81%) and specificity (> 89%). The gyrB sequencing was made in eigth from thirteen misidentified enterobacteria and confirmed 100% the MALDI-TOF results, so the proteomic technique was used as a ?gold standard? for this study. The amino acid decarboxylation test made the most misidentifications and Enterobacter spp was the largest misidentified genus (76.9%, 10/13). E.coli was prevalent (83%, 152/183) in all samples and the bovine milk presented the most enterobacteria diversity. The Salmonella sp wasn?t detected in feces bovine samples and all water samples from different points in the farm presented unacceptable microbiological standards. Was identified enterobacteria in milkers hands and nasal cavity also in the milking machines used on the property. These results aim to contribute significantly to the characterization of the Enterobacteriaceae as well in understanding of its spread in dairy production environment , assisting in need diagnostic of possible agents involved in bovine mastitis as well as to implement properly targeted prophylactic measures. A mastite bovina afeta negativamente a produ??o de leite dificultando a recupera??o dos n?veis de produ??o total das propriedades leiteiras, levando a perdas econ?micas consider?veis. Esta redu??o no percentual da produ??o de leite pode estar associada ao agente patog?nico espec?fico que causou a infec??o, sendo as enterobact?rias frequentemente respons?veis pela mastite ambiental. Estes microrganismos s?o preferencialmente encontrados no habitat normal dos animais como locais que apresentam esterco, urina, barro e camas org?nicas. Os testes fenot?picos est?o entre os m?todos dispon?veis atualmente utilizados para identificar as enterobact?rias; no entanto, eles podem ocasionalmente identitificar erroneamente algumas esp?cies apesar dos m?ltiplos ensaios realizados. Al?m disso, a demora na sua execu??o pode tardar a antibioticoterapia realizada em campo. Por outro lado, a t?cnica de MALDI-TOF MS tem atra?do a aten??o pela sua identifica??o precisa dos v?rios microorganismos em n?vel de esp?cie. No presente estudo, um total de 183 enterobact?rias foram isoladas a partir de amostras de leite (n=47) e fezes colhidas de vacas em lacta??o (n=94); amostras de ?gua (n=23) e na linha de ordenha (n=19) em uma propriedade situada no Rio de Janeiro. A proposta foi utilizar a t?cnica de MALDI-TOF MS como um m?todo eficaz de identifica??o bacteriana de enterobact?rias e descrever a permanencia destes microrganismos no ambiente de produ??o leiteira. A t?cnica prote?mica confirmou 92,9% (170/183) das esp?cies de enterobact?rias identificadas pelos testes bioqu?micos convencionais. O sequenciamento do gene gyrB, realizado em oito das 13 enterobact?rias que apresentaram identifica??o discordante, confirmou em 100% o resultado da t?cnica prote?mica, que foi utilizada como metodologia de refer?ncia no presente estudo. O g?nero Enterobacter foi o mais discordante pelo m?todo bioqu?mico (76,9%, 9/13). A E.coli foi a esp?cie predominante (83%, 152/183) em todas as amostras avaliadas, sendo que o leite bovino apresentou maior diversidade de enterobact?rias. N?o foi detectada a presen?a de Salmonella spp. nas amostras de fezes bovinas e todas as amostras de ?gua dos diferentes pontos de coleta da propriedade apresentaram padr?es microbiol?gicos inaceit?veis. Foram isoladas enterobact?rias das m?os e cavidades nasal dos ordenhadores, bem como nas ordenhadeiras mec?nicas utilizadas na propriedade. Estes dados visam contribuir de forma significativa para a caracteriza??o das enterobacterias bem como para a compreens?o e sua descri??o no ambiente de produ??o leiteira, auxiliando no diagn?stico preciso dos poss?veis agentes envolvidos na mastite bovina bem como na implementa??o de medidas profil?ticas devidamente direcionadas.

Submitted by Sandra Pereira (srpereira@ufrrj.br) on 2017-01-11T12:09:07Z No. of bitstreams: 1 2016 - Joao Bosco Varela.pdf: 1890246 bytes, checksum: 359c15e2f52f2ea873d394cbbfa5479c (MD5) Made available in DSpace on 2017-01-11T12:09:07Z (GMT). No. of bitstreams: 1 2016 - Joao Bosco Varela.pdf: 1890246 bytes, checksum: 359c15e2f52f2ea873d394cbbfa5479c (MD5) Previous issue date: 2016-02-24 The study of ticks and tick-borne disease is increasingly dependent upon the use of molecular biological techniques that are employed in pathogen detection and for the accurate identification of ticks, particularly immature stages. The successful application of molecular methods, principally the polymerase chain reaction (PCR), can only be achieved if the DNA present in the tick was effectively preserved and could be extracted efficiently. The current study compared three fixatives (RNAlater, zinc salts (ZN) and isopropanol) for the ability to preserve the nuclear and mitochondrial (mt) DNA of larvae and nymphs of Amblyomma parvum and larvae of Amblyomma scultptum. DNA was extracted from ticks at times between 72h to 12 months post fixation using a phenol-chloroform procedure and examined using PCR assays for nuclear (internal transcribed spacer 2; ITS2) and mitochondrial (12S rDNA, subunit 1 of cytochrome c oxidase (COI) and D-loop) sequences. The efficiency of amplification was analyzed quantitatively (number of samples which produced amplicon) and qualitatively (relative intensity of bands observed on agarose gels). It was observed that the ITS2 sequence could be amplified in the majority (93,39%, n= 283/303) of the samples, in each of the three fixatives, although qualitative differences were observed, particularly with A. sculptum preserved in ZN. In contrast, fixation in isopropanol effectively abolished the ability to amplify the mitochondrial marker sequences of A. parvum and also resulted in inferior amplification (qualitative), of the D-loop target with A. sculptum. Those effects were observed in samples fixed for as little as 72h. The detrimental effects of isopropanol were also observed in samples extracted using an alkaline lysis method (Hotshot). Samples of A. parvum larvae preserved in RNAlater for 30 months showed an amplification efficacy of 100% in the ITS2 and COI assays, irrespective of the extraction method. Mitochondrial sequences are a central component of the majority of molecular studies of ticks. The findings of this study indicate that isopropanol should be avoided as a fixative for immature stages of ticks. Instead, the use of RNAlater is recommended in order to permit the consistent recovery of amplifiable mtDNA O estudo de carrapatos e doen?as transmitidas por eles ? cada vez mais dependente da utiliza??o de t?cnicas de biologia molecular que s?o empregadas na detec??o de pat?genos, e a acurada identifica??o desses artr?podes, em particular os est?gios imaturos. A aplica??o bem-sucedida dos m?todos moleculares, principalmente, a rea??o em cadeia da polimerase (PCR), s? pode ser alcan?ada se o DNA presente no carrapato foi eficazmente preservado e extra?do de forma eficiente. O estudo comparou tr?s fixadores (RNAlater, sais de zinco (ZN) e isopropanol) avaliando a sua capacidade de preservar DNA mitocondrial (mtDNA) e nuclear de larvas e ninfas de Amblyomma parvum e larvas de Amblyomma scultptum. O DNA foi extra?do dos carrapatos, em tempos entre 72h e 12 meses ap?s a fixa??o por meio da t?cnica de fenol-clorof?rmio e lise alcalina (?Hot Shot?) e examinadas usando ensaios de PCR para sequ?ncias nucleares (espa?ador interno transcrito 2; ITS2) e mitocondriais (12S rDNA, subunidade 1 do citocromo c oxidase (COI) e D-loop). A efici?ncia de amplifica??o foi analisada quantitativamente (n?mero de amostras que produziram ?amplicon?) e qualitativamente (intensidade relativa das bandas observadas em g?is de agarose). Foi observado que a sequ?ncia ITS2 foi amplificada na maioria (93,39%, n= 283/303) das amostras em cada um dos tr?s fixadores, embora tenham sido observadas diferen?as qualitativas, particularmente com A. sculptum preservado em ZN. Em contraste, a fixa??o em isopropanol afetou negativamente a capacidade de amplificar as sequ?ncias dos marcadores mitocondriais de A. parvum e tamb?m resultou na amplifica??o inferior (qualitativa), do alvo D-loop com A. sculptum. Esses efeitos foram observados em amostras fixadas por apenas 72 horas. Os efeitos prejudiciais de isopropanol tamb?m foram observados igualmente em amostras extra?das usando um m?todo de lise alcalina (?HotShot?). As amostras de larvas de A. parvum preservadas em RNAlater durante 30 meses, mostrou uma efic?cia de amplifica??o de 100% nos ensaios para ITS2 e COI, independentemente do m?todo de extra??o usado. Sequ?ncias mitocondriais s?o um componente central da maioria das pesquisas moleculares com carrapatos. Os resultados deste estudo indicam que isopropanol deve ser evitado como um fixador para fases imaturas de carrapatos. Em vez disso, o uso de RNAlater ? recomendado, a fim de permitir a recupera??o consistente de mtDNA amplific?ve

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2017-06-26T17:32:22Z No. of bitstreams: 1 2015 - Thamyris Sampaio Vilela.pdf: 3001387 bytes, checksum: d945149445f8ee2765222f5475c7eaa2 (MD5) Made available in DSpace on 2017-06-26T17:32:22Z (GMT). No. of bitstreams: 1 2015 - Thamyris Sampaio Vilela.pdf: 3001387 bytes, checksum: d945149445f8ee2765222f5475c7eaa2 (MD5) Previous issue date: 2015-02-27 CAPES - Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior CNPq - Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico FAPERJ - Funda??o Carlos Chagas Filho de Amparo ? Pesquisa do Estado do Rio de Janeiro This study aimed to develop a real-time PCR (qPCR) assay for diagnosis of Plasmodium spp. using as target the 18S rDNA and Cyt b genes. A range of 101 blood samples were collected from Gallus gallus in poultry rustic breeds of in Rio de Janeiro, Brazil. The collected bloods were used to prepare blood smears and to extract the deoxyribonucleic acid (DNA) of these samples. There with, molecular protocols were tested, such as the already published conventional PCR (cPCR) and nested PCR (nPCR), designed for Plasmodium spp. In this study two qPCR protocols were developed using primers targeting the Cyt b and 18S rDNA genes. The qPCR detection limit for both genes was 10 copies of the target DNA, which were higher than the detection limit observed in nPCR and cPCR. In qPCR, 69.30% (n = 70/101) samples were positive targeting 18Sr DNA gene and 59.40% (n = 60/101) samples were positive targeting Cyt b gene. In nPCR and cPCR, 54.45% (n = 55/101) and 52.47% (n= 53/101) samples were positive, respectively. In blood smear microscopy, 31 (30.69%) samples were positive. There was no disagreement between the results (p > 0.05) of qPCR for 18Sr DNA and Cyt b genes. Additionally, qPCR was more sensitive than the other techniques discussed, mostly related to blood smear microscopy (p < 0.05). Therefore, the two qPCR developed in the study showed more sensitivity than other techniques and enabled the detection of Plasmodium spp. in poultry even in low parasitemia. Este estudo teve como objetivo desenvolver um ensaio de PCR em tempo real (qPCR) para o diagn?stico de Plasmodium spp. utilizando os genes 18S rDNA e cyt b. Foram coletadas 101 amostras de sangue de aves da esp?cie Gallus gallus de cria??o r?stica ou org?nica no munic?pio de Serop?dica, Rio de Janeiro. Os sangues coletados foram utilizados na prepara??o de esfrega?os de sangue e para extra??o do ?cido desoxirribonucl?ico (DNA) destas amostras. Protocolos de ensaios moleculares foram testados, tal como o PCR convencional (cPCR), nestedPCR (nPCR) e qPCR para Plasmodium spp. Neste estudo dois protocolos de qPCR foram desenvolvidos utilizando oligoiniciadores desenhados com alvo no citocromo b e 18S rDNA. O limite de detec??o para os dois genes qPCR foi de 10 c?pias do alvo de DNA, que foram maiores do que o limite de detec??o observado em nPCR e cPCR. Em qPCR, 69,30% (n = 70/101) amostras foram positivas com alvo no gene 18SrDNA e 59,40%(n = 60/101) amostras positivas com alvo no gene cyt b. Em nPCR e cPCR, 54,45% (n = 55/101) e 52,47% (n = 53/101) amostras foram positivas, respectivamente. Em microscopia de esfrega?o de sangue, 31 (30,69%) amostras foram positivas. N?o houve discord?ncia entre os resultados (p> 0,05) de qPCR para os genes 18SrDNA e cyt b.. Al?m disso, qPCR foi mais sens?vel do que as outras t?cnicas discutidas, principalmente relacionado com a microscopia ?ptica (p

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2019-11-13T18:21:08Z No. of bitstreams: 1 2015 - Michele da Costa Pinheiro.pdf: 4006380 bytes, checksum: f9fb9ac88f68d69607bb4b8d249fa6fd (MD5) Made available in DSpace on 2019-11-13T18:21:08Z (GMT). No. of bitstreams: 1 2015 - Michele da Costa Pinheiro.pdf: 4006380 bytes, checksum: f9fb9ac88f68d69607bb4b8d249fa6fd (MD5) Previous issue date: 2015-02-23 CAPES FAPERJ It is extremely important the knowledge about ecological characteristics in different biomes and how each population behaves to adversities of the environment they live in. It has been observed that some tick species widely distributed may have genetic and / or biological differences between populations. The tick Amblyomma parvum has a wide distribution Neotropical with an extensive list of hosts. To try to elucidate these gaps on the subject, studies were developed in Laborat?rio de Ixodologia and Biologia Molecular in Departamento de Parasitologia Animal in Universidade Federal Rural do Rio de Janeiro. When comparing the life cycle of this parasite from three colonies of different locations (Pantanal Sul-matogrossense, Cerrado, Goias and Chaco, Argentina) it was observed that the complete cycle of A. parvum was completed in an average period of 104.2; 99.6 and 100.7 days for the Pantanal, Chaco and Cerrado respectively. We conclude that the biological parameters of the three populations are quite similar. It is also not fully understood as terrestrial arthropods survive the temporary flooding following heavy rains then the purpose of the second chapter was to compare the effect of immersion in the life cycle in order to respond to water stress can affect and/or changes in the reproductive efficiency. It can be observed that the Pantanal population was more sensitive to immersion even though the observation ecdysis in group 96 hours. Despite this in group 48 of the Pantanal population there was no continuation of the cycle while the Chaco and Cerrado populations continued its cycle even after the nymphs remain submerged 48 hours. It can be observed that from 72 hours immersion causes a deleterious effect on this species. The results reported here suggest that the ability to survive underwater for a period is a additional aspect of the biology of this species of tick. With respect to the molecular analysis was demonstrated in literature genetic divergence (3.0 to 3.7%) of A. parvum Argentinean and Brazilian. However, analysis of a single gene is not robust enough to make inferences about the significance of the genetic differences between different populations, so new sequences of molecular markers obtained from A. parvum belonging to different locations in Argentina and Brazil have been evaluated. Results for the genes 12S and COI there was a significant difference point of to consider them as distinct species in South America, 3.8% and 7.1% respectively. But when we observe the results for the ITS2 gene there was not enough difference to characterize different species. View of these biological and molecular results are recommended further analysis with new samples of new locations in order to elucidate this question about the possibility of this kind in South America be different species. ? de suma import?ncia o conhecimento sobre caracter?sticas ecol?gicas em diferentes biomas e como cada popula??o se comporta frente a adversidades do meio em que vive. Tem-se observado que algumas esp?cies de carrapatos amplamente distribu?dos podem apresentar diferen?as gen?tica e/ou biol?gicas entre as popula??es. O carrapato Amblyomma parvum possui uma ampla distribui??o Neotropical com uma lista extensa de hospedeiros. Para tentar elucidar essas lacunas sobre o assunto, estudos foram desenvolvidos nos Laborat?rios de Ixodologia e Biologia Molecular do Departamento de Parasitologia Animal da Universidade Federal Rural do Rio de Janeiro. Ao comparar o ciclo biol?gico deste parasito proveniente de tr?s col?nias de distintas localidades (Pantanal Sul-matogrossense, Cerrado goiano e Chaco argentino) observou-se que o ciclo completo de A. parvum foi completado em um per?odo m?dio de 104,2; 99,6 e 100,7 dias para os biomas Pantanal, Chaco e Cerrado respectivamente. Conclui-se que os par?metros biol?gicos das tr?s popula??es s?o bastante semelhantes. Tamb?m n?o ? totalmente compreendido como artr?podes terrestres sobrevivem ?s inunda??es tempor?rias que seguem fortes chuvas ent?o o objetivo do segundo cap?tulo foi comparar o efeito da imers?o sob o ciclo de vida a fim de responder como o estresse h?drico pode afetar e/ou alterar a efici?ncia reprodutiva. Pode-se observar que a popula??o Pantanal demonstrou ser mais sens?vel em rela??o ? imers?o mesmo tendo a observa??o de ecdise no grupo 96 horas. Apesar disso no grupo 48 horas da popula??o Pantanal n?o houve continua??o do ciclo enquanto que as popula??es Chaco e Cerrado continuaram seu ciclo mesmo depois que as ninfas permanecerem 48 horas submersas. Pode-se observar que a partir de 72 horas a imers?o causa um efeito delet?rio para essa esp?cie. Os resultados aqui relatados sugerem que a capacidade de sobreviver submersos por um per?odo ? um aspecto adicional da biologia dessa esp?cie de carrapato. No que diz respeito ? an?lises moleculares na literatura foi demonstrado diverg?ncia gen?tica (3,0 a 3,7%) entre A. parvum argentinos e brasileiros. No entanto, a an?lise de apenas um gene n?o ? suficiente para fazer infer?ncias robustas sobre a signific?ncia das diferen?as gen?ticas entre diferentes popula??es, ent?o novas sequ?ncias de marcadores moleculares obtidas a partir de A. parvum pertencentes a diferentes localidades da Argentina e Brasil, foram analisadas. para o gene 12S e COI houve uma diferen?a significativa a ponto de consider?-las como esp?cies distintas na Am?rica do Sul, 3,8% e 7,1% respectivamente. Mas ao observarmos os resultados para o gene ITS2 n?o houve diferen?a suficiente para caracterizar esp?cies diferentes. Diante desses resultados biol?gicos e moleculares s?o recomend?veis novas an?lises com novas amostras de novas localidades, a fim de elucidar essa d?vida sobre a possibilidade desse g?nero da Am?rica do Sul ser esp?cies diferentes.

Submitted by Sandra Pereira (srpereira@ufrrj.br) on 2019-06-18T15:57:31Z No. of bitstreams: 1 2015 - Bruna de Azevedo Ba?ta.pdf: 4031086 bytes, checksum: 4030be45fc06aa6f754bc07f162f640a (MD5) Made available in DSpace on 2019-06-18T15:57:31Z (GMT). No. of bitstreams: 1 2015 - Bruna de Azevedo Ba?ta.pdf: 4031086 bytes, checksum: 4030be45fc06aa6f754bc07f162f640a (MD5) Previous issue date: 2015-02-20 Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico, CNPq, Brasil In the first chapter two new strains of Anaplasma marginale derived from the Serop?dica, RJ were isolated and characterized in tick cells line of Ixodes scapularis (IDE8), enabling new studies in the diagnosis with the use of local strains. In the second chapter the first lines of embryonic cells of Rhipicephalus (Boophilus) microplus from Brazilian strains were developed and characterized. The strains of A. marginale were from blood of two naturally infected calves belonging to the Universidade Federal Rural do Rio de Janeiro (UFRRJ), Serop?dica. After isolation, the strains were characterized morphologically by light microscopy and molecularly by nested PCR of msp1? gene. Two strains of A. marginale were isolated and named as AmRio1 and AmRio2. Small colonies were observed in cytospin smears stained with Giemsa?s 1:10, of each of the isolates, 60 days after culture initiation. Analysis of msp1? microsatellite in both strains resulted in a single genotype, genotype E. The amino acid sequence of one MSP1? tandem repeat from the strain AmRio1 resulted in a new sequence (named 162) with one amino acid change. The results of our phylogenetic analysis demonstrated that A. marginale strains from Brazil and Argentina formed two large clusters. In establishing of the strains of R. (B.) microplus, the ticks were from strain kept calves in Parasitic Diseases Laboratory, UFRRJ, Serop?dica. After 13 days of laying, the pool of eggs was sterilized, macerated, filtered and centrifuged at 200 xg for 10 minutes. The pellet was suspended in Leibotiz?s L15 medium and divided into 6 flasks of 25 cm2, of which 3 were contaminated, one was deposited in the cell bank of the Pirbright Institute and others two were characterized in the present study (BMEBR1 and BMEBR2 lines) through molecular analysis of the 16S rDNA gene (PCR), growth curve and epifluorescence and light microscopy. Sixty days in culture, the cells began to show confluent aspect, reaching the formation of the monolayer. There was a complete similarity (100%) of new lines BMEBR1 and BMEBR2 with R. (B.) microplus in Brazil (EU918178.1), Costa Rica (EU918179.1), Paraguay (EU918180.1), among others. Both strains showed lower growth than the R. (B.) microplus line from Costa Rica, BME/CTVM2. The BMEBR1 line has increased phagocytic ability that the BMEBR2 and BME/CTVM2 lines. In addition, these strains have acidification characteristics, lipid corpuscle production and oxidative stress different from each other. The new strains were frozen and defrosted successfully, and were stable with respect to morphology and growth rate. Thus, these lines can be used in future studies to propagation of new strains of A. marginale isolated. No primeiro cap?tulo foram isoladas e caracterizadas duas novas cepas de Anaplasma marginale oriundas do munic?pio de Serop?dica, RJ, em linhagem de c?lulas de carrapato Ixodes scapularis (IDE8), possibilitando novos estudos no diagn?stico com a utiliza??o de cepas locais. No segundo cap?tulo foram desenvolvidas e caracterizadas as primeiras linhagens de c?lulas embrion?rias de Rhipicephalus (Boophilus) microplus oriundas de cepa brasileira. As cepas de A. marginale foram obtidas a partir de sangue de dois bezerros naturalmente infectados pertencentes ? Universidade Federal Rural do Rio de Janeiro (UFRRJ), Serop?dica. Ap?s isolamento, as cepas foram caracterizadas morfologicamente, atrav?s da microscopia de luz e molecularmente atrav?s do semi-nested PCR do gene msp1?. Duas cepas de A. marginale foram isoladas e nomeadas como AmRio1 e AmRio2. Pequenas col?nias foram observadas em l?minas de citocentr?fuga coradas com Giemsa 1:10, de cada isolado, 60 dias ap?s o in?cio da cultura. An?lises de uma repeti??o em tandem de MSP1a da cepa AmRio1 resultou em uma nova sequ?ncia (nomeada de 162) com um amino?cido diferente. O resultado da an?lise filogen?tica demonstrou que as cepas de A. marginale a partir do Brasil e Argentina formam dois grandes clusters. Para o estabelecimento das linhagens de R. (B.) microplus, os carrapatos foram provenientes de cepa mantida em bezerros no Laborat?rio de Doen?as Parasit?rias, UFRRJ, Serop?dica. No 13? dia de postura, a massa de ovos foi esterilizada, macerada, filtrada e centrifugada a 200 xg por 10 minutos. O ?pellet? foi suspenso em meio Leibovitz?s L15 e dividido em 6 frascos de 25 cm2, dos quais 3 contaminaram, um foi depositado no banco de c?lulas do Instituto Pirbright e outros 2 foram caracterizados no presente estudo (linhagens BMEBR1 e BMEBR2) atrav?s da an?lise molecular do gene 16S DNAr (PCR), curva de crescimento e microscopia de luz e epifluoresc?ncia. Com 60 dias de cultivo, as c?lulas passaram a apresentar um aspecto confluente, caracterizando a forma??o da monocamada. Existiu uma completa similaridade (100%) das novas linhagens BMEBR1 e BMEBR2 com R. (B.) microplus do Brasil (EU918178.1), da Costa Rica (EU918179.1), do Paraguai (EU918180.1), entre outros. Ambas as cepas apresentaram crescimento menor que a linhagem de R. (B.) microplus BME/CTVM2 oriunda da Costa Rica. A linhagem BMEBR1 possui maior capacidade fagoc?tica que as linhagens BMEBR2 e BME/CTVM2. Al?m disso, essas linhagens possuem caracter?sticas de acidifica??o, produ??o de corp?sculo lip?dico e estresse oxidativo diferentes entre si. As novas linhagens foram congeladas e descongeladas com sucesso e apresentaram-se est?veis com rela??o ?s caracter?sticas morfol?gicas e de taxa crescimento. Desta forma, estas linhagens podem ser utilizadas em futuros estudos na propaga??o das novas cepas de A. marginale isoladas.

CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior Hemosporídeos de aves silvestres são considerados importantes agentes etiológicos e podem causar danos às populações dos hospedeiros. O presente estudo teve como objetivos: determinar a prevalência e a parasitemia para Plasmodium spp. e Haemoproteus spp. de Passeriformes da Mata Atlântica de Minas Gerais, por meio de dois métodos de diagnóstico, microscopia e nested PCR e caracterizar morfologicamente a espécie Plasmodium (Haemamoeba) lutzi Lucena, 1939. Amostras de sangue foram obtidas por punção da veia braquial para preparação de esfregaços que foram fixados em metanol e corados em Giemsa. Foram amostradas 237 aves pertencentes a 18 famílias e 56 espécies da ordem Passeriformes provenientes do IBAMA e capturadas no Jardim Botânico da Universidade Federal de Juiz de Fora (JB-UFJF). A prevalência para Plasmodium spp. foi 63,63% em aves do IBAMA e 26,25% em aves do JB-UFJF, apenas três indivíduos encontraram-se parasitados por Haemoproteus spp., sendo dois (2,5%) provenientes do IBAMA e um (0,62%) do JB-UFJF. A parasitemia média encontrada para Plasmodium spp./Haemoproteus spp. foi de 0,08 (± 0,08) no IBAMA e 0,05 (± 0,04) no JB-UFJF. O maior grau de parasitemia média foi registrado nas espécies Turdus albicollis, Zonotrichia capensis (0,22%), Turdus rufiventris (0,18% ± 0,29), Gnorimopsar chopi (0,1% ± 0,07) no IBAMA e Turdus rufiventris (0,17 % ± 0,13), Arremon semitorquatus (0,11% ± 0,13) no JB-UFJF. Foi identificada em uma das aves (A. semitorquatus), a presença da espécie Plasmodium (H.) lutzi, a qual foi morfologicamente caracterizada. A prevalência registrada é considerada alta, entretanto verificou-se uma baixa parasitemia, o que pode estar relacionado à evolução crônica da infecção. Infecções agudas por Plasmodium spp. levam ao aparecimento de sinais clínicos com expressão rápida dos sintomas, sendo que nos casos mais graves pode ocorrer a morte do hospedeiro. Visto a elevada prevalência, levantamentos acerca de infecções por hemoparasitos em aves silvestres podem ajudar na elaboração de estratégias de profilaxia e tratamento de doenças parasitárias para a conservação das espécies. Hemosporidian wild birds are considered important etiological agents and can cause damage to populations of hosts. The present study aimed to: ¹determine prevalence and parasitemia for Plasmodium spp. and Haemoproteus spp. Passeriformes of the Atlantic Forest of Minas Gerais, through two diagnostic methods, microscopy and nested PCR; and caracterize morphologically the species Plasmodium (Haemamoeba) lutzi Lucena, 1939. Blood samples were obtained by puncturing the brachial vein for preparation of smears, fixed in methanol and stained with Giemsa. 237 birds belonging to 18 families and 56 species of the order Passeriformes from IBAMA were sampled and captured in the Botanical Garden of the Universidade Federal de Juiz de Fora (BG-UFJF). The prevalence of Plasmodium spp. in birds of IBAMA was 63.63% and 26.25% in birds of BG-FUJF, only three individuals were infected by Haemoproteus spp., two (2.5%) from IBAMA and one (0.62 %) of BG-FUJF. The average parasitemia found for Plasmodium spp./ Haemoproteus spp. was 0.08 (± 0.08) at IBAMA and 0.05 (± 0.04) in BG-UFJF. The highest degree of mean parasitaemia was recorded in the species Turdus albicollis, Zonotrichia capensis (0.22%), Turdus rufiventris (0.18% ± 0.29), Gnorimopsar chopi (0.1% ± 0.07) at IBAMA and Turdus rufiventris (0.17% ± 0.13), Arremon semitorquatus (0.11% ± 0.13) in BG-UFJF. Was identified in one of the birds (A. semitorquatus) by means of morphology, the presence of Plasmodium (H.) lutzi species, which was characterized morphologically. Registered prevalence is considered high, however a low parasitaemia was found, which may be related to chronic course of infection.. Acute infections with Plasmodium spp. lead to the onset of clinical signs with rapid symptom expression, and in severe cases death canoccurat the host. Since the high prevalence surveys about hemoparasites infections in wild birds can help in developing strategies for prophylaxis and treatment of parasitic diseases to species conservation.

Submitted by Celso Magalhaes (celsomagalhaes@ufrrj.br) on 2019-10-16T13:23:09Z No. of bitstreams: 1 2014 - ?sis Daniele Alves Costa Santolin.pdf: 3929893 bytes, checksum: 503fc63d41e038dfae557b3e332f69b3 (MD5) Made available in DSpace on 2019-10-16T13:23:09Z (GMT). No. of bitstreams: 1 2014 - ?sis Daniele Alves Costa Santolin.pdf: 3929893 bytes, checksum: 503fc63d41e038dfae557b3e332f69b3 (MD5) Previous issue date: 2014-09-22 Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES Ticks parasites of wild birds may serve as vectors of pathogens of major importance for wildlife and public health. Thus, the present study examined the prevalence and intensity of infestation of ticks captured from wild birds in three regions of the Atlantic Forest, with different levels of human impact, located in Nova Igua?u, State of Rio de Janeiro, Brazil, with the objective of highlighting the possible relationship between prevalence of infestation and preservation of forest areas in the region. Collections were conducted from May 2011 to April 2012, resulting in the capture of 625 birds represented by 98 species divided into 26 families and seven orders, which equated to 14.2% of the richness of bird species in the state. A total of 17.44% (n= 109, 109/625) of the captured birds were parasitized by 458 ticks, of which 88.2% (n = 404, 404/458) were in the larval stage (LL) and 11.8% (n= 54, 54/458) in the nymphal stage. Among the ticks collected, 99% (n = 453) were represented by the genus Amblyomma: Amblyomma longirostre (LL = 345; NN = 34), Amblyomma nodosum (NN = 13), Amblyomma calcaratum (NN = 3), Amblyomma varium (NN = 2); Amblyomma coelebs (NN = 1), Amblyomma naponense (LL = 1), Amblyomma ovale (LL = 1) and Amblyomma sp. (LL = 53). The genus Haemaphysalis was represented by four larvae and a single nymph, corresponding to 1% of the ticks collected. In addition, new species of host birds were reported in Brazil, along with an unprecedented case of super infestation with larvae of A. longirostre on Xiphorhynchus guttatus. Molecular analysis (sequencing of mitochondrial 16S rDNA), provided validation of morphological identification of larvae of A. longirostre at the species level without to need to apply methods of clarification commonly used. The detection and identification of pathogens and their tick hosts are increasingly dependent upon molecular biological methods, which have as their first step the extraction of DNA from the tick and the microorganisms contained therein. In an attempt to standardize the extraction of DNA from ticks and their asscoaited microbiota, the present study compared six methods of DNA extraction (Boiling in sterile water, InstaGene?, QIAamp? DNA Mini Kit, Phenol:chloroform, Diatomaceous earth and modified Hotshot). To date, samples subjected to extraction with the kit and Phenol:chloroform have demonstarted superior results (in terms of quantity and quantity of extracted DNA), when compared to the other methods. In silico analysis of sequences encoding the gltA, htrA and ompB genes of tick associated rickettsial species provided the basis for a novel PCR-RFLP method which allowed differential identification of the Rickettsia species reported in Brazil. The method was evaluated using larvae and nymphs of Amblyomma longirostre, Amblyomma ovale and Amblyomma varium collected from birds captured in this study. The bacterium "Candidatus Rickettsia amblyommii" was identified in 100% (17/17) of A. longirostre larvae examined, while the other two species of ticks were universally PCR negative. The basic method employs the restriction endonucleases MspI and RsaI, can be performed during a single working day. It offers a convenient and cost effective means to perform large scale analysis of tick populations, and should be of benefit to researchers who lack the financial or technical resources necessary for nucleotide sequence based identification. In conclusion, this study expands the understanding of the ecological relationships between ticks and birds in a major biome of the Neotropics, with new perspectives in the context of molecular techniques applied to investigations of parasitism involving tick parasites of wild birds Carrapatos parasitos de aves silvestres podem servir como vetores deagentes patog?nicos de grande import?ncia para a sa?de silvestre e coletiva. Assim, o presente estudo analisou a preval?ncia e intensidade de infesta??o de carrapatos em aves silvestres capturadas em tr?s regi?es de Mata Atl?ntica, com diferentes n?veis de antropiza??o, situada em Nova Igua?u, estado do Rio de Janeiro, Brasil, com o objetivo de avaliar a rela??o entre a preval?ncia de infesta??o e ?reas preservadas na regi?o. As capturas foram realizadas no per?odo de maio de 2011 a abril de 2012 onde foram capturadas 625 aves representadas por 98 esp?cies divididas em 26 fam?lias e sete ordens, totalizando 14,2% da riqueza das esp?cies de aves do estado. O total de 17,44% (n=109; 109/625) das aves capturadas estavam parasitadas por 458 carrapatos, sendo 88,2% (n=404; 404/458) no est?gio de larva (LL) e 11,8% (n=54; 54/458) no est?gio de ninfa. Dentre os ixod?deos coletados, 99% (n=453) eram do g?nero Amblyomma: Amblyomma longirostre (LL= 345; NN= 34), Amblyomma nodosum (NN= 13), Amblyomma calcaratum (NN= 3), Amblyomma varium (NN= 2); Amblyomma coelebs (NN=1), Amblyomma naponense (LL= 1), Amblyomma ovale (LL=1) e Amblyomma sp. (LL= 53). O g?nero Haemaphysalis sp. foi representado por quatro larvas e uma ?nica ninfa, correspondendo a 1% dos carrapatos coletados. Al?m disso, foram relatadas novas esp?cies de aves hospedeiras no Brasil, o caso in?dito de superinfesta??o por A. longirostre em Xiphorhynchus guttatus. An?lises moleculares (sequenciamento do gene mitocondrial 16S DNAr), permitiram a valida??o da identifica??o morfol?gica ao n?vel de esp?cie de larvas de A. longirostre sem a necessidade de aplica??o de m?todos de clarifica??o comumente utilizados. A detec??o e identifica??o de agentes patog?nicos e de seus hospedeiros carrapatos dependem cada vez mais de m?todos moleculares, que tem por princ?pio a extra??o do DNA do carrapato e do microrganismo abrigado por ele. Na tentativa de estandardiza??o da extra??o do DNA de ixod?deos e de sua microbiota associada, o presente estudo comparou seis m?todos de extra??o de DNA (Fervura em ?gua est?ril, InstaGene?, QIAamp? DNA Mini Kit, Fenol:clorof?rmio, Terra de diatom?cea e Hotshot modificado). At? o momento, amostras submetidas ? extra??o com Kit e Fenol:clorof?rmio apresentam melhores resultados (em termos de quantidade e qualidade do DNA extra?do) quando comparado aos outros m?todos. An?lise in silico de sequ?ncias que codificam os genes gltA , htrA e ompB de carrapatos associados ? esp?cies de rickettsias revelaram ser a base para um novo m?todo de PCR-RFLP, que permitiu a identifica??o diferencial de esp?cies de Rickettsia encontradas no Brasil. O m?todo foi avaliado utilizando larvas e ninfas de Amblyomma longirostre, Amblyomma ovale e Amblyomma varium coletadas nas aves capturadas neste estudo. A bact?ria ?Candidatus Rickettsia amblyommii? foi identificada em 100% (17/17) dos A. longirostre examinados, enquanto que as outras duas esp?cies de carrapatos foram universalmente PCR negativas. O m?todo b?sico emprega as endonucleases de restri??o MspI e Rsal e pode ser realizada no decurso de um ?nico dia de trabalho. Este oferece um meio conveniente e rent?vel para executar a an?lise em grande escala de popula??es de carrapatos, e deve ser um benef?cio para os pesquisadores que n?o disp?em de recursos financeiros ou t?cnicos necess?rios para a identifica??o baseada no sequenciamento de nucleot?deos. Em conclus?o, este estudo amplia o conhecimento das rela??es ecol?gicas entre ixod?deos e a avifauna em um importante bioma da regi?o Neotropical, com novas perspectivas no ?mbito das t?cnicas moleculares aplicadas a inqu?ritos de parasitismo que envolve carrapatos parasitos de aves silvestres

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2020-02-14T18:30:40Z No. of bitstreams: 1 2014 - Rubens Fabiano Soares Prado.pdf: 2042043 bytes, checksum: 143c18335d78655e1170899efb5d0c6d (MD5) Made available in DSpace on 2020-02-14T18:30:40Z (GMT). No. of bitstreams: 1 2014 - Rubens Fabiano Soares Prado.pdf: 2042043 bytes, checksum: 143c18335d78655e1170899efb5d0c6d (MD5) Previous issue date: 2014-02-26 The tick-borne diseases in addition to have a worldwide distribution, related to the dispersion of these vectors, and a complex epidemiologic network, making them difficult to control and prevention, cause major impacts on human and animal health. Brazil, the second largest herd of horses in the world, has the Theileria equi as a major hemoparasites transmitted by ticks to such animals, causing serious barriers to equideocultura in the country, which is considered endemic. Moreover, some aspects of its epidemiology, as the real vector of the disease in the country, are still not well understood. Regarding zoonotic diseases that the animals are involved, an emerging borreliosis, the Baggio - Yoshinari Syndrome, has drawn attention because horses seropositive for Borrelia spp. has been evidenced in our country, suggesting possible involvement of these animals in its epidemiology. Thus, this study aimed to perform serological diagnosis and assess the risk factors associated with seropositivity for Theileria equi and Borrelia spp. in the herd of horses for military use in the municipality of Resende, state of Rio de Janeiro. Samples of 174 horses for military use were tested by indirect ELISA immunoassay for Theileria equi and Borrelia burgdorferi strain G39/40. The dependent variable (seroreactivity) was tested through bivariate analysis for the association as the independent variables (gender, age, racial definition, group management, creation time in AMAN, system creation, origin of the horse, the presence of infestation by Dermacentor nitens, by Amblyomma cajennense, level of tick infestation and presence of tick infestation). The variables that were associated in bivariate analysis (p < 0.25) were included in the logistic regression model for multivariate analysis. It was found that the prevalence Borrelia spp. was 29.89%, and the T. equi prevalence was 74.14 %. Multivariate analysis demonstrated that the presence of tick infestation is associated (p < 0.05) with seropositivity for Borrelia spp. in horses (OR 3,45; CI 1,2 ? 9,7), and the creation time in AMAN greater than 15 years associated with seropositivity for T.equi (OR 5,70; CI 1,23 - 22,1). From the results obtained it can be inferred that ticks are related in the transmission of Borrelia spp in the herd studied, with their probable vectors. Further evidence of circulating Borrelia spp. in horses reinforces their role as sentinels, warning of the occurrence of human cases. Regarding Theileria equi, it was found that the area is endemic for the disease with a high prevalence of asymptomatic carriers, evoking care and preventive measures in the introduction of horses, coming from areas unaffected or enzootic instability, in the herd. As doen?as transmitidas por carrapatos al?m terem um ampla distribui??o mundial, relacionada ? dispers?o destes vetores, e uma rede epidemiol?gica complexa, tornando-as de dif?cil controle e preven??o, causam grandes impactos na sa?de humana e animal. O Brasil, com o terceiro maior rebanho de equ?deos do mundo, tem como um dos principais agentes transmitidos por carrapatos aos equ?deos Theileria equi. Tal agente ? considerado end?mico e vem causando entraves ? equideocultura nacional. Al?m disso, alguns aspectos de sua epidemiologia, como o artr?podo vetor da doen?a no pa?s, ainda n?o foram elucidados. No que tange ?s doen?as zoon?ticas em que os equ?deos est?o envolvidos, a S?ndrome de Baggio-Yoshinari, uma borreliose emergente, tem chamado a aten??o, pois t?m sido evidenciados equinos soropositivos para Borrelia spp. em nosso pa?s, sugerindo o envolvimento destes animais na epidemiologia desta doen?a. O objetivo deste estudo foi realizar o diagn?stico sorol?gico e avaliar os fatores de risco associados ? soropositividade para Theileria equi e para Borrelia spp. no rebanho de equinos de uso militar do munic?pio de Resende, estado do Rio de Janeiro. Amostras de 174 equinos de uso militar foram testadas atrav?s do ensaio de adsor??o imunoenzim?tica indireta iELISA para Theileria equi e Borrelia burgdorferi cepa G39/40. A vari?vel dependente (sororreatividade) foi testada, atrav?s de an?lise bivariada, quanto ? associa??o ?s vari?veis independentes (g?nero, idade, defini??o racial, grupo de manejo, tempo de cria??o na AMAN, sistema de cria??o, origem do equino, presen?a de infesta??o por Dermacentor nitens, presen?a de infesta??o por Amblyomma cajennense, grau de infesta??o por carrapatos, e presen?a de infesta??o por carrapatos). As vari?veis que apresentaram associa??o na an?lise bivariada (p

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2019-10-11T19:25:33Z No. of bitstreams: 1 2014 - Juliana Macedo Raimundo.pdf: 1424493 bytes, checksum: a82bd96caa7755d7bbcf76de2d28b759 (MD5) Made available in DSpace on 2019-10-11T19:25:33Z (GMT). No. of bitstreams: 1 2014 - Juliana Macedo Raimundo.pdf: 1424493 bytes, checksum: a82bd96caa7755d7bbcf76de2d28b759 (MD5) Previous issue date: 2014-02-21 CAPES Mycoplasma spp. and Cytauxzoon felis are hemoparasites with potential to cause anemia and other debilitating conditions in domestic cats. The aim of this study was to detect by molecular technique hemoplasma and Cytauxzoon felis infection in cats from Metropolitan Area of Rio de Janeiro, as well as linking it to the hematological data. From a total of 197 cats sampled, 45 (22.8%) were positive by Real-Time Polymerase Chain Reaction (qPCR) to Mycoplasma spp. and 22 (11.2%) have shown structures compatible with haemoplasma organisms on blood smear. The infection frequency of hemoplasma species by PCR, using primers specific to hemoplasma species, was 4.6%, 4.6% and 11.7% for Mycoplasma haemofelis (Mhf), ?Candidatus M. turicensis (CMt) and ?Candidatus Mycoplasma haemominutum? (CMhm), respectively. Therefore ?Candidatus M. haemominutum? was the specie with the highest incidence in the cats studied. Co-infection between Mhf and CMhm (1.0%), Mhf and CMt (0.5%) and CMt and CMhm (0.5%) have been observed, as well as triple co-infection in one animal (0.5%). No difference of Mycoplasma spp. distribution was observed between animals from Baixada Fluminense districts (23.9%) and Rio de Janeiro (21.3%). However, there was a higher distribution of positive cats for ?Candidatus M. haemominutum? and ?Candidatus M. turicensis? in the district of Rio de Janeiro. Mycoplasma haemofelis infection was higher in Baixada Fluminense, whose cats had five times more chance to be infected by this agent (p> 0.05). Mycoplasma spp., M. haemofelis and ?Candidatus M. haemominutum infections were higher in the summer, on the other hand, ?Candidatus M. turicensis showed a peak of infection in the winter, these findings were not statistic different (p>0.05). Regarding hematological changes associated with M. haemofelis infection, macrocytic hypochromic anemia (p0.05). Although no statistical association was found, adult cats were more likely to be infected by hemoplasma (p>0.05). All animals were negative for Cytauxzoon felis by PCR, although piroplasma forms were seen in erythrocytes of two animals. The results demonstrate the occurrence of three species of haemotropic mycoplasmas and piroplasmas at Metropolitan Area of Rio de Janeiro and denote their potential to cause anemia. Consequently, the knowledge about the disease and the factors related to its occurrence are required for the adoption of preventive action aiming at animal and man health. Os hemoparasitos Mycoplasma spp. e Cytauxzoon felis s?o micro-organismos com potencial para causar anemia e demais condi??es debilitantes em felinos dom?sticos. O objetivo do presente estudo foi detectar molecularmente a infec??o por hemoplasmas e Cytauxzoon felis em gatos da Regi?o Metropolitana do Rio de Janeiro, associando-a aos dados hematol?gicos. Dos 197 gatos amostrados, 45 (22,8%) foram positivos pela Rea??o em Cadeia da Polimerase em Tempo Real (qPCR) para Mycoplasma spp. e 22 (11,2%) apresentaram estruturas epieritroc?ticas sugestivas na pesquisa direta em esfrega?o sangu?neo. A frequ?ncia de infec??o das esp?cies de hemoplasma pela PCR, utilizando oligonucleot?deos esp?cie-espec?ficos, foi 4,6%, 4,6% e 11,7% para M. haemofelis (Mhf), ?Candidatus M. turicensis (CMt) e ?Candidatus Mycoplasma haemominutum? (CMhm), respectivamente. ?Candidatus M. haemominutum? constitui, portanto, a esp?cie de maior ocorr?ncia na popula??o de gatos estudada. Foram identificadas coinfec??es entre Mhf e CMhm (1,0%), Mhf e CMt (0,5%) e CMhm e CMt (0,5%), assim como coinfec??o tripla em um animal (0,5%). N?o foi observada diferen?a na distribui??o de Mycoplasma spp. entre os animais provenientes dos munic?pios da Baixada Fluminense (23,9%) e Rio de Janeiro (21,3%). No entanto, constatou-se maior distribui??o de gatos positivos para ?Candidatus M. haemominutum? e ?Candidatus M. turicensis? no munic?pio do Rio de Janeiro. J? a infec??o por M. haemofelis foi maior na Baixada Fluminense, cujos gatos apresentaram cinco vezes mais chance de se infectarem por este agente (p>0,05). Os picos de infec??o por Mycoplasma spp., M. haemofelis e ?Candidatus M. haemominutum? foram maiores no ver?o, por outro lado, ?Candidatus M. turicensis demonstrou pico de infec??o no inverno, tais achados n?o foram significativos estatisticamente (p>0,05). No tocante ?s altera??es hematol?gicas associadas ? infec??o por M. haemofelis, constatou-se anemia predominantemente do tipo macroc?tica hipocr?mica (p0,05). Embora sem associa??o estat?stica, gatos adultos apresentaram mais chances de se infectarem por hemoplasmas (p>0,05). Todos os animais foram negativos para Cytauxzoon felis pela PCR, embora tenham sido evidenciadas formas de piroplasma em hem?cias de dois animais. Os resultados demonstram a ocorr?ncia das tr?s esp?cies de micoplasmas hemotr?ficos e de piroplasmas na regi?o metropolitana do Rio de Janeiro e denotam seu potencial como causador de anemia. Consequentemente, o conhecimento da doen?a bem como dos fatores relacionados ? sua ocorr?ncia s?o necess?rios para a ado??o de medidas preventivas visando ? sa?de animal.

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2019-10-15T18:09:40Z No. of bitstreams: 1 2014 - Gabriela Lopes Vivas Vitari.pdf: 1102825 bytes, checksum: f158aa2a469fa226216c23d0fc3a18ac (MD5) Made available in DSpace on 2019-10-15T18:09:40Z (GMT). No. of bitstreams: 1 2014 - Gabriela Lopes Vivas Vitari.pdf: 1102825 bytes, checksum: f158aa2a469fa226216c23d0fc3a18ac (MD5) Previous issue date: 2014-02-20 CAPES - Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior CNPq - Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico FAPERJ - Funda??o Carlos Chagas Filho de Amparo ? Pesquisa do Estado do Rio de Janeiro Equine piroplasmosis stands out among the diseases that affect the majority of horse herds in Brazil, and is caused by protozoans, of intraerythrocytic location, named Theileria equi and Babesia caballi . This study aimed to determine the rate of natural infection by T. equi and B. caballi in ticks collected from pastures frequented by horses and/or cattle in Serrana and Itaguai micro-regions in state of Rio de Janeiro, Brazil. Furthermore, the other objective was to evaluate the possible associated factors to rate of infected ticks collected in different properties, according to the characteristics of each micro-region studied. The tick collections were performed in 28 properties in the studied areas between January and August 2013. Ticks were collected from pastures by dragging flannel method over the vegetation, CO2 traps and manual collection after viewing. The ticks collected were identified, counted, separated by life stage and pools of ticks samples were formed. Genomic DNA was extracted and the samples were analysed by real time PCR and nested PCR. Four thousand seven hundred sixteen (4.716) ticks were collected, 84.8% (n=3.997) of Amblyomma genus, 10.9% (n=514) of Dermacentor genus and 4.3% (n=205) of Rhipicephalus genus. From the samples collected, 4.276 were larvae, 431 nymphs and 9 adults. In 92.3% (N=13) properties of Itaguai micro-region was found the presence of ticks in the grazing area to animals, differing significantly (p

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2019-09-24T20:04:17Z No. of bitstreams: 1 2014 - Renata Lins da Costa.pdf: 1119547 bytes, checksum: 89be1c9c66ce6fcfb90b3b14055cfb42 (MD5) Made available in DSpace on 2019-09-24T20:04:17Z (GMT). No. of bitstreams: 1 2014 - Renata Lins da Costa.pdf: 1119547 bytes, checksum: 89be1c9c66ce6fcfb90b3b14055cfb42 (MD5) Previous issue date: 2014-02-18 CAPES This work was developed in properties with creations of horses in the micro-regions of Itaguai and Serrana, both located in the state of Rio de Janeiro. The study aimed to evaluate the role of molluscs and trematodes in Neorickettsia risticii cycle in creations located in the micro-region of Itagua? and analyze the frequency of antibody against N. risticii in horses from Serrana micro-region, standing out the possible associated factors with the seropositivity of animals. Among thirteen properties studied in Itaguai micro-region were collected 410 specimens of limnic molluscs using hand net adapted by fine screen. The molluscs were exposed to the incandescent light (60 w) during 2-4 hours for research of trematodes larval forms. Subsequently, proceeded to molecular detection of N. risticii by Polymerase Chain Reaction in real time (qPCR) in DNA samples of molluscs as in samples of trematodes larval. Many taxons of molluscs were found: 71.70% (294/410) represented by Melanoides tuberculata; 10.97% (45/410) by Pomacea sp.; 3.65% (15/410) by Drepanotrema anatinum; 6.58% (27/410) by Biomphalaria tenagophila; 0.24% (1/410) by B. straminea; 5.60% (23/410) by Physa acuta and 1.21% (5/410) by Family Hydobriidae. A frequency of 3.17% (13/410) of molluscs parasited by trematodes was observed. According to the morphometric and biological characteristics observed in trematodes were classified the following cercariae types: Megalourous cercariae, Pleurolophocercous cercariae and Furcocercous cercariae. No amplification of the target DNA for N. risticii was observed in any DNA samples of molluscs and trematodes analyzed. Regarding to sero-epidemiological study of N. risticii in horses of Serrana micro-region, blood samples from 354 animals were collected, residing at 49 properties of region, by jugular venipuncture in tubes of 5mL without anticoagulant. Blood samples were subjected to centrifugation to obtainment of serums which were stored in freezer at -20?C until analysis moment. For detection of antibodies against N. risticii was used the Immunofluorescence Antibody Test (IFAT), and the samples were considered positive when reacted to N. risticii at 1:50 dilution. Between the municipalities of Petropolis and Teresopolis, was found a frequency of 5.9% of serologically reactive horses. Among the possible factors analyzed, it was found that gender, breed, age, zootechnical and sanitary management, presence of snails and flooded areas did not present association with seropositivity of horses by N. risticii (p>0.05). The region of origin or provenance of the animal was the only factor associated (p0,05). A regi?o de origem ou proced?ncia do animal foi o ?nico fator associado (p

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2018-10-19T18:19:28Z No. of bitstreams: 1 2013 - Andresa Guimar?es.pdf: 1221110 bytes, checksum: 80ca8363650edbb042caea761cb85083 (MD5) Made available in DSpace on 2018-10-19T18:19:28Z (GMT). No. of bitstreams: 1 2013 - Andresa Guimar?es.pdf: 1221110 bytes, checksum: 80ca8363650edbb042caea761cb85083 (MD5) Previous issue date: 2013-08-21 CAPES CNPq Studies of domestic cats are little discussed in Brazil, especially regarding hemoparasita research where cats can be carriers of the agent and contribute to infection of man. Anaplasmataceae agents are gram-negative bacteria found in leukocytes and platelets, in which morulae characteristic of the infection is observed. The transmission of these agents to humans and animals occurs by arthropod vectors. Sorologic diagnostic methods are emphasized, but recent studies using molecular techniques demonstrate greater sensitivity and specificity, with better characterization of the agent. The present study aims to diagnose domestic cats in Rio de Janeiro naturally infected by agents Anaplasmataceae (Ehrlichia sp., and Anaplasma sp.) by serological and molecular methods, as well as evaluate hematological changes associated with these infections to guide veterinarians in the diagnosis of these diseases. Samples of 216 animals from clinics located in cities in the metropolitan region of Rio de Janeiro were used. The research of antibodies IgG anti-E. canis was performed by Immunofluorescence Assay (IFAT) and molecular detection of Ehrlichia sp. by nested Polymerase Chain Reaction (PCR) based on 16S rRNA gene. The results demostrate that 18 (8.3%) animals were positive in the direct detection by blood smear, 57 (26.4%) seropositive and 37 (17.1%) positive by nested-PCR for Ehrlichia sp., being 91 (42.1%) positive in at least one technique. The diagnosis of A. platys was performed by real-time PCR (qPCR) and nested-PCR based on 16S rRNA and gltA genes, respectively. Seventeen animals (7.9%) showed inclusions in platelet, eight (3.7%) were positive in qPCR, two (0,9%) in nested-PCR and 23 (10.6%) cats were considered positive for A. platys in at least one technique. The detection of A. phagocytophilum was performed by qPCR based on gene msp2, however, positive animals were not observed. The most common hematologic findings in animals positive for agents Anaplasmataceae were thrombocytopenia, leukocytosis, neutrophilia, regenerative left shift, anemia and hyperproteinemia. Only one animal showed co-positivity for both agents. Regarding the associated factor, only age interfered with positivity for Ehrlichia sp., in wich younger IFAT positive animals were less prone to infection, unlike that observed in the nested-PCR in which the younger had higher positivity. Only two positive samples in the nested-PCR for E. canis were subjected to sequencing. One sequence demonstrated 100% similarity with isolates of Ehrlichia sp. and E. canis and another showed 99% similarity with isolates of Ehrlichia sp. ocelot and Ehrlichia sp. little-spotted-cat. The two positive samples in the nested-PCR for A. platys demonstrated 100% similarity with isolates of A. platys reported in advanced. The present work demonstrates the circulation of agents Anaplasmataceae in domestic cats in Rio de Janeiro. Estudos com felinos dom?sticos s?o pouco abordados no Brasil, principalmente em rela??o ? pesquisa de hemoparasitoses, em que os gatos podem ser portadores do agente e contribuir para infec??o do homem. Os agentes Anaplasmataceae s?o bact?rias gram-negativas que s?o encontradas em leuc?citos, plaquetas ou eritr?citos, formando m?rulas, caracter?sticas de sua infec??o. A transmiss?o desses agentes para o homem e animais ocorre principalmente por vetores artr?podes. Como m?todo diagn?stico ressalta-se a sorologia, por?m estudos recentes utilizando diagn?stico molecular demonstram maior sensibilidade e especificidade, com melhor caracteriza??o do agente. O presente estudo tem como objetivo diagnosticar felinos dom?sticos do Rio de Janeiro infectados naturalmente por agentes Anaplasmataceae (Ehrlichia sp. e Anaplasma sp.) por meio de m?todos sorol?gico e molecular, bem como avaliar as altera??es hematol?gicas associadas a estas infec??es. Foram utilizadas amostras de 216 animais, provenientes de cl?nicas localizadas em cidades da Regi?o Metropolitana do Rio de Janeiro. A pesquisa de anticorpos IgG anti-E. canis foi realizada pela Rea??o de Imunofluoresc?ncia Indireta (RIFI) e a detec??o molecular de Ehrlichia sp., pela nested Rea??o em Cadeia da Polimerase (nested-PCR) baseada no gene 16s rRNA. Os resultados demonstraram que 18 (8,3%) animais foram positivos na detec??o direta pelo esfrega?o sangu?neo, 57 (26,4%) soropositivos e 37 (17,1%) positivos na nested-PCR para Ehrlichia sp., sendo 91 (42,1%) considerados positivos para Ehrlichia sp. em pelo menos uma t?cnica. O diagn?stico de A. platys foi efetuado pela PCR em tempo real (qPCR) e nested-PCR baseada nos genes gltA e 16S rRNA, respectivamente. Dezessete animais (7,9%) apresentaram inclus?es em plaquetas, oito (3,7%) foram positivos na qPCR, dois (0,9%) na nested-PCR e 23 (10,6%) animais foram considerados positivos para A. platys em pelo menos uma t?cnica. A pesquisa de A. phagocytophilum foi realizada por meio da qPCR baseada no gene msp2; no entanto, n?o foram observados animais positivos. Os achados hematol?gicos mais frequentes nos animais positivos para agentes Anaplasmataceae foram trombocitopenia, leucocitose, neutrofilia, desvio ? esquerda regenerativo, anemia e hiperproteinemia. Apenas um animal apresentou co-positividade para ambos os agentes. Em rela??o aos dados associados, somente a idade interferiu na positividade para Ehrlichia sp., sendo na RIFI os animais mais jovens menos predispostos ? infec??o, ao contr?rio do observado na nested-PCR em que os mais jovens apresentaram maior ?ndice de positividade. Apenas duas amostras positivas na nested-PCR para E. canis foram submetidas ao sequenciamento, sendo que uma sequ?ncia apresentou 100% de similaridade com isolados de Ehrlichia sp. e E. canis e a outra demonstrou 99% de similaridade com isolados de Ehrlichia sp. ocelot e Ehrlichia sp. little-spotted-cat. As duas amostras positivas na nested-PCR para A. platys apresentaram 100% de similaridade com isolados de A. platys previamente descritos. O presente trabalho demonstra, portanto, a ocorr?ncia de agentes Anaplasmataceae na popula??o de felinos dom?sticos do Rio de Janeiro.