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Abstract Background Currently, various zoonotic diseases are classified as emerging or reemerging. Because equids have a direct relationship with various vectors, they are possibly more frequently exposed to zoonotic agents than are humans. The undeniable importance of diseases such as human granulocytic anaplasmosis, spotted fever, and leishmaniasis for both public and animal health, as well as the possibility of equids acting as sources, reservoirs, or even sentinels for these pathogens, justifies the detection of their frequency and factors associated with infection in equids from northeastern Brazil. Methods Blood samples were collected from 569 equids (528 horses, 33 donkeys, and 8 mules), 516 from a rural area and 53 from an urban area. Pathogen detection was carried out as follows: Borrelia spp. and Rickettsia spp., serological analysis; Leishmania spp., serological analysis and polymerase chain reaction (PCR); Anaplasma phagocytophilum, PCR. Determination of associated factors was carried out through generalized linear models. Results The frequencies of positivity for the pathogens observed in equids were as follows: Borrelia spp., 13.9% (79/569); Leishmania spp., 3.5% (20/569); Rickettsia spp. 33.4% (190/569). Regarding factors associated with infection, male sex was associated with protection against Borrelia spp.; donkeys and mules were associated with protection against Rickettsia spp., while a younger age was a risk factor. The infection of A. phagocytophilum was not detected in the sampled population. Co-infection was detected in 5.1% (29/569) of the animals. Conclusions Most of the studied pathogenic agents are present in the prospected area, indicating a possible risk for both human and animal health. This demonstrates that equids can be considered important sentinels in the assessment of pathogens with zoonotic potential in the region.

Abstract The aim of this study was to determine the frequency and factors associated to Babesia caballi, Theileria equi and Trypanosoma evansi in naturally infected equids from the northeast Brazil. Blood samples from 569 equids (528 horses, 8 mules, and 33 donkeys) were collected and tested for the presence of DNA of each of these protozoan parasites by PCR. Generalized linear models were used to evaluate risk factors associated with the infection. The frequency of T. equi infection was 83.5% (475/569) - 84.3% in horses, and 73.2% in donkeys and mules. The results of the final model indicated that age (senior group) and animal species (mule and donkey group) were protective factors against this pathogen. The frequency of B. caballi infection was 24.3% (138/569) - 23.5% in horses and 34.1% in donkeys and mules. Age (adult and senior group) was considered a protective factor against B. caballi infection whereas animal species (donkey and mule group) were considered a risk factor for the infection. Trypanosoma evansi infection was not detected in any of animals. Our results suggest that equids from the area studied may be infected earlier in life with the etiological agents of equine piroplasmosis and become asymptomatic carriers.

Chagas disease, caused by the flagellate protozoan Trypanosoma cruzi, is an anthropozoonosis of great importance for public health. Several species of mammals are reservoirs for this parasite, including dogs. The objective of this work was to verify the natural infection by T. cruzi in the population of dogs of the municipality of Ituberá, Bahia. A total of 392 domiciled dogs from all districts of the city were evaluated; five milliliters of blood was collected from the dogs for molecular diagnosis. T. cruzi DNA was amplified through the polymerase chain reaction (PCR); the primers P35 and P36, which amplify a fragment of 330 bp, were used. The PCR products were subjected to 2% agarose gel electrophoresis containing Sybr (Invitrogen). Of the 392 dogs evaluated, only 2 (0.51%) animals, one male and one female, tested positive for T. cruzi. This study concluded that there are dogs naturally infected by T. cruzi in the municipality of Ituberá-Bahia and that this finding is an alert to veterinarians, health professionals, and local health authorities that their own dogs can act as reservoirs of the disease.

Toxoplasmosis, leishmaniasis, and ehrlichiosis are zoonoses that have cosmopolitan distribution and cause important clinical changes in dogs and humans. Dogs are important in the epidemiology and transmission of Leishmania infantum and Ehrlichia canis and serve as indicators of environmental contamination by the transmission of Toxoplasma gondii. The aim of this study was to detect antibodies against T. gondii, Leishmania spp., and E. canis in dogs from Bahia and identify the risk factors associated with infections. Blood samples were collected from 353 dogs from several municipalities of Bahia and the sera were tested using reaction of indirect immunofluorescence (RIFI) for T. gondii and Leishmania spp. and enzyme-linked immunosorbent assay (ELISA) for E. canis. The data were tabulated and analyzed using EPI-INFO 7.1.5 program. Of 353 samples analyzed, 144 (40.8%) were positive for T. gondii (antibody titer 16-4,096) and 92 (26.1%) were seropositive for Leishmania spp. (antibody titer 40-160). The seropositivity for E. canis was the highest as compared to that of the other agents. Furthermore, 184 (52.1%) samples were positive for antibodies against E. canis. Sixteen (4.5%), 71 (20.1%), and 25 (7.1%) samples were co-infected with T. gondii and Leishmania spp., T. gondii and E. canis, and E. canis and Leishmania spp., respectively, while 20 (5.7%) dogs presented co-seropositivity for all three etiological agents. Risk factors such as age, sex, habitat (stray/not stray), and region of dog’s origin were analyzed; age ( ? 1 year) was significantly associated with T. gondii (p = 0.03) and E. canis (p = 0.02) infection and stray animals were more exposed to T. gondii (p = 0.03) and Leishmania spp. (p = 0.01) infection. The results confirmed the heterogeneous distribution of the three etiological agents in dogs from Bahia and identified animals with co-infections.

A Doença de Chagas, causada pelo protozoário flagelado Trypanosoma cruzi, é uma antropozoonose de grande importância para a saúde pública. Diversas espécies de mamíferos são reservatórios do parasita, incluindo o cão. O objetivo deste trabalho foi verificar a infecção natural por T. cruzi na população de cães do município de Ituberá, Bahia. Foram avaliados 392 cães domiciliados, em todos os bairros do município, dos quais foram coletados 5mL de sangue para realização do diagnóstico molecular. A amplificação do DNA de T. cruzi foi através da Reação em Cadeia da Polimerase (PCR), com os primers P35 e P36, que amplificam um fragmento de 330 pb. Os produtos das PCRs foram submetidos à eletroforese em gel de agarose a 2% contendo Sybr (Invitrogen®). Dos 392 cães avaliados, apenas 2 (0,51%) animais, um macho e uma fêmea, foram positivos no diagnóstico molecular de T. cruzi. Conclui-se com esse estudo que há cães naturalmente infectados pelo T. cruzi no município de Ituberá-Bahia e que este achado constitui um alerta aos veterinários, profissionais da saúde e autoridades sanitárias locais, cujos cães podem atuar como reservatórios da doença.

A toxoplasmose, a leishmaniose e a erliquiose são zoonoses que possuem distribuição cosmopolita e causam importantes alterações clínicas em cães e em seres humanos. Cães são importantes na epidemiologia e transmissão de Leishmania infantum e Ehrlichia canis, e podem ser indicadores de contaminação ambiental na transmissão de T. gondii. Objetivou-se com esse estudo detectar anticorpos IgG anti-Toxoplasma gondii, Leishmania spp. e Ehrlichia canis em cães do estado da Bahia, bem como identificar fatores de risco associados as infecções. Amostras de sangue de 353 cães foram coletadas em diversos municípios deste estado e os soros foram submetidos à reação de imunofluorescência indireta (RIFI) para T. gondii e Leishmania spp. e ensaio imunoenzimático indireto (ELISA) para E. canis. Os dados foram tabulados e analisados utilizando-se o programa EPIINFO 7.1.5. Para o agente etiológico T. gondii, das 353 amostras coletadas, 144 (40,8%) foram positivas, com títulos de anticorpos variando de 16 a 4096. Para Leishmania spp., 92 (26,1%) amostras mostraram soropositividade, com títulos de anticorpos variando de 40 a 160. A soropositividade para E. canis foi a maior em relação aos outros agentes. Dos animais amostrados, 184 (52,1%) tiveram anticorpos anti-E. canis, no ELISA. Das 353 amostras analisadas, verificou-se co-soropositividade em 16 (4,5%) animais para T. gondii e Leishmania spp.; 71 (20,1%) animais foram positivos para T. gondii e E. canis; 25 (7,1%) para E. canis e Leishmania spp.; e 20 (5,7%) cães apresentaram co-soropositividade para os três agentes etiológicos. Foram analisados como fatores de risco a idade, o sexo, se o animal era errante ou não, e a mesorregião de origem dos cães. Dentre os fatores de risco analisados, a idade ( ? 1ano) foi significativa para T. gondii (p = 0,03) e E. canis (p = 0,02) e animais errantes se apresentaram mais expostos para T. gondii (p = 0,03) e Leishmania spp. (p = 0,01). Os resultados comprovam a presença dos três agentes etiológicos nos municípios estudados no estado da Bahia, distribuídos de maneira heterogênea e apresentando animais com coinfecções.

In order to verify the Trypanosoma cruzi infection in domestic domiciled dogs in a rural endemic area from the south region of the State of Bahia, Polymerase Chain Reaction (PCR) were performed using S35 and S36 primers in 272 dogs living in the district of Vila Operaria, in the municipality of Buerarema. All animals were clinically evaluated; 2.5 mL of blood were collected through venipuncture for the performance of molecular tests. None of these animals showed clinical signs of the illness and only two were identified with the DNA parasite. This result is the first report of natural infection by T. cruzi in domestic dogs in southern Bahia.Com o objetivo de verificar a infecção por Trypanosoma cruzi em cães domésticos domiciliados em área rural e endêmica do sul da Bahia, foi realizada a Reação em Cadeia da Polimerase (PCR), utilizando-se os iniciadores S35 e S36 em 272 cães domiciliados no distrito da Vila Operária, cidade de Buerarema. Todos os animais foram avaliados clinicamente e, posteriormente, foram coletados 2,5 mL de sangue por punção venosa para realização do diagnóstico molecular. Nenhum dos animais apresentou manifestação clínica da doença e, em apenas dois foram identificados DNA do parasito. Esse resultado é o primeiro relato de infecção natural por T. cruzi em cães domésticos no sul baiano.

The aim of this work was to study the clinical disorders and risk factors of canine ehrlichiosis in Ilhéus and Itabuna, Bahia, and compare different diagnostic methods. Blood samples were collected from 200 dogs. Each dog was clinically examined. A questionnaire was used to evaluate the risk factors. The blood samples were analyzed using the Dot-ELISA test; hematometry, platelet counts and searches for morulae on blood smears were performed. Nested PCR was carried out on 50 serologically positive samples and 50 negative samples. Three positive PCRs were sequenced. Thirty-six percent were serologically positivity and 5.5% from blood smears. The animals were anemic and thrombocytopenic. Presence of ticks and living in areas on the urban periphery were considered to be risk factors (p < 0.05). Nested PCR identified 11 positive dogs of which nine were serologically positive and two were negative. The DNA sequencing was consistent with the presence of Ehrlichia canis.Objetivou-se com este trabalho estudar as alterações clínicas, fatores de risco da ehrlichiose canina nos municípios de Ilhéus e Itabuna, Bahia, e comparar diferentes métodos de diagnóstico. Amostras de sangue foram coletadas de 200 cães e cada animal foi examinado clinicamente. Foi preenchido um questionário para avaliar os fatores de risco. As amostras de sangue foram analisadas pelo teste Dot-ELISA e foram realizadas hematimetria, contagem de plaquetas e procura de mórulas em esfregaço de sangue. Nested-PCR foi realizada em 50 amostras positivas e 50 negativas na sorologia. Três amostras PCRs positivas foram seqüenciadas. Foi encontrado 36,0% de positividade na sorologia e 5,5% nos esfregaços sanguíneos. Os animais apresentavam anemia e trombocitopenia. Ter carrapatos e residir em áreas suburbanas foram considerados fatores de risco (p < 0,05). A Nested-PCR identificou 11 cães positivos, sendo 9 com sorologia positiva e 2 negativos. O sequenciamento de DNA foi compatível com a presença de Ehrlichia canis.

Twenty Nelore and twenty F1 cattle (½ Red Angus, ½ Nelore), average age 12 months were compared in pastured raised at southern Bahia, in relation to production characteristics: initial weight (IW), final weight (FW), weight gain, average daily gain (ADG) and total weight gain (WG), infection by endoparasites and heat tolerance (HT). To analyze the effect of genetic group, it was stabilished effects that could influence these characteristics and performed variance analysis using the program SAS. The results showed that IW, FW, ADG and WG of crossbred were statistically different in comparison to zebu cattle. Related to infection by endoparasites and heat tolerance there were no statistical differences between genetic groups.

Foram comparados 20 animais da raça Nelore e 20 animais F1 (½ Red Angus ½ Nelore), com idade média de 12 meses, criados a pasto no Sul da Bahia, em relação às características de produção: peso inicial (PI), peso final (PF), ganho de peso médio diário (GMD) e ganho de peso total (GT); infecção por endoparasitos e, tolerância ao calor (ITC). Para análise de efeito do grupo genético, considerou-se fixo todos os outros efeitos que poderiam influenciar essas características e fez-se a análise de variância utilizando-se o programa SAS. Os resultados do estudo mostraram que PI, PF, GMD e GT dos animais cruzados foram, estatisticamente, superiores aos dos animais zebuínos. Em relação à infecção por endoparasitos e tolerância ao calor não foram observadas diferenças significativas entre os grupos genéticos.

Ehrlichiosis is a zoonotic disease that is caused by bacteria of the genus Ehrlichia. The aims of this study were to detect the presence of Ehrlichia spp. in the blood of dogs in Ituberá, Bahia, and to compare the sensitivities and specificities of blood smear, serological, and molecular examinations. Furthermore, this study identified factors associated with exposure to the agent in dogs in this locality. Blood samples were collected from 379 dogs and submitted for indirect immunofluorescent assay and polymerase chain reaction testing for the detection of Ehrlichia spp. antibodies and DNA, respectively. Additionally, a peripheral blood smear was obtained from the ear tip for parasite identification. Of the 379 animals, 12.4%, 32.7%, and 25.6% were identified as positive on the blood smear, serological, and molecular tests, respectively. The dogs positive in one of the three techniques were considered exposed (46.9%). Younger dogs and rural habitat were protective factors and presence of ticks and contact with other dogs were the risk factors associated with exposure to the agent. It was concluded that dogs of Ituberá have high positivity for Ehrlichia spp. and that the diagnostic methods used for detection are complementary.

In order to verify the Trypanosoma cruzi infection in domestic domiciled dogs in a rural endemic area from the south region of the State of Bahia, Polymerase Chain Reaction (PCR) were performed using S35 and S36 primers in 272 dogs living in the district of Vila Operaria, in the municipality of Buerarema. All animals were clinically evaluated; 2.5 mL of blood were collected through venipuncture for the performance of molecular tests. None of these animals showed clinical signs of the illness and only two were identified with the DNA parasite. This result is the first report of natural infection by T. cruzi in domestic dogs in southern Bahia.

Direct diagnoses were made by using - blood smears and nested PCR (nPCR) tests on 309 blood samples from crossbred dairy cattle in the municipality of Ibicaraí, Bahia. From diagnostic blood smear slides, the observed parasitic frequencies were 31.1% for Anaplasma marginale and 20.4% for Babesia sp. From nPCR diagnoses, they were 63% for A. marginale, 34% for Babesia bigemina and 20.4% for Babesia bovis. There were significant differences (P

Abstract In Brazil, Leishmania infantum mainly affects humans and dogs. The state of Bahia presents many dogs that are positive for this parasite. Despite the importance of epidemiology in this region, there are still very few studies that have assessed the genetic characteristics of L. infantum. The aim of this study is to investigate the genetic variability of L. infantum isolated identified in naturally infected dogs, in order to verify occurrence of subpopulation of this parasite in the different biomes existing in the state of Bahia. Thirty-two samples of L. infantum were analyzed, which were obtained isolated in dogs from the Mata Atlântica (rainforest), Caatinga (semi-arid scrub forest), and Cerrado (a vast tropical savannah eco-region) Bahia municipalities’ biomes. All animals presented with clinical changes suggestive of Leishmania spp. and they exhibited positive reactions to serological tests. kDNA analysis with RFLP markers revealed the presence of genetic variability and gene flow in subpopulations of L. infantum; samples from the Mata Atlântica areas were genetically more similar to those from the areas of Caatinga and they were less likely to resemble those of the Cerrado. This data may be used to investigate the dissemination of parasite in the canine population of state of Bahia.

RESUMO. Rocha J.M., Carvalho F.S., Oliveira H.C., Carlos R.S.A., Carneiro P.L.S., Albuquerque G.R. & Wenceslau A.A. [Molecular diagnosis of Ehrlichia canis in dogs with clinical signs, hematological and biochemical changes in the Municipality of Ilhéus, Bahia.] Diagnóstico molecular de Ehrlichia canis em cães com alterações clínicas, hematológicas e bioquímicas no município de Ilhéus, Bahia. Revista Brasileira de Medicina Veterinária, 38(4):345-352, 2016. Programa de Pós-Graduação em Ciência Animal, Universidade Estadual de Santa Cruz, Rodovia Jorge Amado, Km 16, Salobrinho, Ilhéus, BA 45662-900, Brasil. E-mail: wenceslau@uesc.br Objetivou-se com este trabalho identificar DNA de Ehrlichia canis em cães com alterações clínicas e ou hematológicas condizentes com a infecção, e identificar possíveis fatores de risco à infecção no município de Ilhéus, uma área endêmica à infecção. Foram coletadas 558 amostras de sangue de cães. Essas amostras foram divididas em frascos com e sem anticoagulante e foram confeccionados esfregaços sanguíneos para pesquisa direta na lâmina. Durante o momento da coleta os animais foram submetidos a exame clínico completo e avaliação para presença de carrapatos. O sangue com anticoagulante foi utilizado para realização de hemograma completo e testes moleculares para detecção de E. canis e o sem anticoagulante para os exames bioquímicos de Ureia, Creatinina e ALT. Antes da análise molecular e em lâmina, foi realizada uma triagem, onde foram selecionados animais que apresentassem sinais clínicos, hematológicos e bioquímicos sugestivos da infecção. Após essa triagem, foram selecionados 297 cães para a pesquisa de Ehrlichia sp. e E. canis. Dos animais avaliados pela metodologia molecular 85,52% (254/297) foram positivos para o gênero e 80,13% (238/297) foram positivos para a espécie E. canis pelo Nested–PCR. A visualização de mórulas nas lâminas foi verificada em 28,16% (84/297) de animais e todos também foram positivos pelo Nested-PCR. Dos animais positivos para E. canis, 25,39 % apresentaram caquexia, 17,46% apresentaram palidez em mucosas, 15,87% apresentaram petéquias e 1,58% apresentaram epistaxe, e outros 38,09% (24/63) apresentaram mais de uma das alterações concomitantes. Animais que apresentaram alterações em sinais clínicos foram estatisticamente significativos (p=0,02) em relação à positividade para E. canis. Com relação às alterações hematológicas 80,13% dos cães estavam infectados, sendo estas estatisticamente significantes (p=0,001). Os animais com anemia apresentaram significância estatística (p=0,001), porém, não foi constatada significância nos animais trombocitopênicos (p=0,28). Quando se avaliou animais que possuíam anemia em associação com trombocitopenia, os resultados foram significativos estatisticamente (p0,05), porém 48,82% (145/297) animais apresentaram controle carrapaticida anterior ao momento da coleta e, este dado foi significante (p=0,003). Com relação a avaliação bioquímica os resultados não foram estatisticamente significativos (p>0,05) para a infecção. Foram detectados pela análise molecular 19,04% de carrapatos infectados das 42 amostras de carrapatos coletados. Conclui-se que a região tem alta infecção por E. canis e os animais neste estudo, possivelmente estavam cursando com a fase crônica da infecção, no município de Ilhéus, BA.