The objective of this study was to identify and characterize species of Eimeria in broiler chickens using traditional morphological and pathological plus molecular (DNA amplification) diagnostic methodologies. Using a combination of those techniques it was possible to identify the presence of multiple circulating species in the flock as well as higher frequencies for some of them, especially Eimeria praecox and Eimeria maxima, which were identified in 100% of the flocks. The frequencies of the other species were Eimeria mitis and Eimeria necatrix (93.3%), Eimeria tenella (76,7%), Eimeria acervulina (56.7%) and Eimeria brunetti (16.7%). However using the lesion score, the most common species were E. maxima (46.7%), E. acervulina (30%), E. tenella (23.3%), and E. necatrix (10%). E. brunetti and E. praecox were not identified by using lesion score. DNA amplification had detection sensitivity for Eimeria species in the field samples of at least 20 oocysts. The implementation of DNA amplification as a routine diagnostic technique in aviaries can assist Eimeria population.

Bovine babesiosis and anaplasmosis complex: diagnosis and evaluation of the risk factors from Bahia, BrazilComplexo tristeza parasitária bovina: diagnóstico e avaliação dos fatores de risco na Bahia, Brasil AbstractDirect 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 < 0.01) between the two diagnostic methods (nPCR and blood smear slides). The compliance obtained from the kappa test was 0.41 and 0.48 for A. marginale and Babesia sp., respectively. The tick samples from the six farms analyzed using nPCR were only positive for A. marginale. Evaluation of the risk factors relating to the presence of ticks and the age of the animals showed that there was a significant association (P < 0.01) with the frequency of animals infected with both pathogens. Therefore, under the conditions studied, nPCR proved to be a good tool for diagnosing the agents of the bovine babesiosis and anaplasmosis complex because of its sensitivity and specificity in comparison with blood smears. The municipality of Ibicaraí is an area with endemic prevalence of bovine babesiosis and anaplasmosis confirmed by nPCR and A. marginale is the main agent of the disease.Keywords: Bovine Babesiosis and Anaplasmosis Complex, blood smear, nPCR, risk factors. ResumoRealizou-se o diagnóstico direto por esfregaço sanguíneo e nested PCR (nPCR) em 309 amostras de sangue de bovinos mestiços leiteiros provenientes do município de Ibicaraí, Bahia. A frequência observada no diagnóstico por lâminas de esfregaço sanguíneo foi 31,1% para Anaplasma marginale e 20,4% para Babesia sp. Enquanto que no diagnóstico por nPCR foi 63% para A. marginale, 34% para Babesia bigemina e 20,4% Babesia bovis. Verificaram-se diferenças significativas (P<0,01) na comparação entre os dois métodos de diagnósticos (nPCR e esfregaço sanguíneo). A concordância ao teste KAPPA obtida foi de 0,41 e 0,48 para A. marginale e Babesia sp., respectivamente. As amostras de carrapatos das seis propriedades analisadas por nPCR foram positivas apenas para A. marginale. Na avaliação dos fatores de risco verificou-se que a presença de carrapato e idade dos animais apresentaram associação significativa (P<0,01) com a frequência de animais infectados por ambos os patógenos analisados por nPCR. Portanto, nas condições estudadas, a nPCR revelou-se uma boa ferramenta para diagnóstico dos agentes do complexo tristeza parasitária bovina (TPB) devido a sensibilidade e especificidade, quando comparado ao esfregaço sanguíneo. O município de Ibicaraí

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.Keywords: Diagnosis, dog, Ehrlichia canis, Indirect Immunofluorescence, nested PCR. ResumoErliquiose é uma doença zoonótica causada por bactérias do gênero Ehrlichia. O objetivo desse estudo foi detectar a presença de Ehrlichia spp. no sangue de cães em Ituberá-BA, e comparar as sensibilidades e especificidades do esfregaço sanguíneo, e testes sorológico e molecular. Além disso, esse estudo identificou fatores associados com a exposição ao agente em cães desta localidade. Amostras de sangue foram coletadas de 379 cães e submetidas à Reação de Imunofluorescência Indireta e Reação em Cadeia de Polimerase para detecção de anticorpos e DNA de Ehrlichia spp., respectivamente. Adicionalmente, sangue periférico de ponta de orelha foi coletado para identificação do parasita. Dos 379 animais, 12,4%, 32,7% e 25,6% foram identificados como positivos no esfregaço sanguíneo, teste sorológico e molecular, respectivamente. Cães positivos em uma das três técnicas foram considerados expostos (46,9%). Cães mais novos e hábitat rural foram fatores de proteção e presença de carrapatos e contato com outros cães foram os fatores de risco associados à exposição ao agente. Foi concluído que, os cães do município de Ituberá têm alta positividade para Ehrlichia spp. e que os diferentes métodos diagnósticos utilizados para sua detecção são complementares.

ABStRACt. The objective of the present article was an epidemiological and molecular study of Ehrlichia canis in dogs of Ilhéus and Itabuna in Bahia, as well as an evaluation of associated risk factors. Blood samples were collected from 153 dogs and DNA was extracted and analyzed by the nested-polymerase chain reaction, using one pair of primers to detect Ehrlichia bacteria and another pair to detect the presence of E. canis. Of the 153 animals, 12 (7.8%) were polymerase chain reaction-positive for E. canis, indicating the presence of the parasite in dogs of the Ilhéus-Itabuna microregion. The associated risk factors were exposure to tick-infested habitats and the fact that the dogs lived in the countryside.

Bovine tick-borne disease (TBD) constitutes a worldwide group of diseases that result in great losses for dairy and beef cattle. With regard to the epidemiological profile of the diseases, the importance of transplacental transmission is still not very well understood. The aim of this study was to determine the transplacental transmission of TBD agents (Anaplasma marginale, Babesia bovis and B. bigemina) in a herd of dairy cattle that had been naturally infected in an area of enzootic stability in northeastern Brazil. Blood for serology of the three agents was collected from cows within 120 days of gestation and serology, haemogram and nPCR assays were performed after birth. Blood was collected from the calves within 3h of birth, and haemogram and nPCR assays were performed in all animals. Pre-colostrum serology was achieved in 34 animals. The Student's t-test was used to compare the haemogram results between animals that were positive and negative for the haemoparasites. The cows were seropositive for all agents in at least one of the examinations. We detected 15 cases of vertical transmission of A. marginale, 4 of B. bovis and 2 of B. bigemina in the 60 cows. In infected animals, co-infection was detected for A. marginale and B. bovis in 1 of 60 calves, and a triple infection was detected in one other calf. Fatal neonatal anaplasmosis was observed in 1 of 15 calves, in which death occurred within 24h of birth. From the results, we concluded that transplacental transmission of TBD agents occurs, including in cases of co- and triple-infection. Such transplacental transmission can cause neonatal death, increasing the importance of this form of epidemiological transmission and suggesting its role as a cause of undiagnosed neonatal death.