Maria Aparecida da Gloria Faustino

Molecular survey and genetic characterization of tick-borne pathogens in dogs in metropolitan Recife (north-eastern Brazil)

To identify DNA of the main tick-borne pathogens in dogs from Recife (Brazil), polymerase chain reactions were carried out on blood samples of dogs treated at the Veterinary Hospital of the Universidade Federal Rural de Pernambuco from March 2007 to June 2008. The detection of DNA was performed using specific primers. Amplicons were analyzed through electrophoresis and sequencing. A phylogenetic tree was constructed using the UPGMA method, revealing that the sequences were closely related to those of strains from other geographic regions. Among the 205 blood samples analyzed, 48.78% was positive for Anaplasma platys; 38.04% was positive for Ehrlichia canis; 7.31% was positive for Babesia canis vogeli; and 0.49% was positive for Hepatozoon canis and Mycoplasma haemocanis. Coinfection of two or three pathogens was found in 23.9% (49/205) of the dogs. The subspecies B. canis vogeli was identified. Infection by H. canis and M. haemocanis is reported for the first time in dogs in the state of Pernambuco (Brazil). The data indicate that the main tick-borne pathogens in dogs in this region are E. canis and/or A. platys, followed by B. canis vogeli.

SURVEY OF CANINE HEARTWORM IN THE CITY OF RECIFE, PERNAMBUCO, BRAZIL

Six hundred and eleven random-source dogs (338 male, 273 female) one year of age or older, from six sections of the city of Recife, Pernambuco, were examined antemortem for circulating microfilariae Dirofilaria immitis and Dipetalonema reconditum adult heartworm (D. immitis) antigen, and examined postmortem for adult heartworms. The prevalence of heartworm infection was 2.3% (14/611), as determined by necropsy for adult worms, and 1% (6/611) had circulating microfilariae of D. immitis; thus, 57.1% of the heartworm-infected dogs had occult infections. The results of serological testing indicated that 1.3% (8/611) of the dogs were positive for adult heartworm antigen. A total of 42 (6.9%) of the dogs had microfilariae of D. reconditum; 40 of these had only D. reconditum and two additional dogs had microfilariae of both species, D. immitis and D. reconditum.

Molecular detection of Anaplasma platys in a naturally-infected cat in Brazil

Following the accidental finding of inclusion bodies similar to Anaplasma platys in a stained blood smear from a cat, DNA analysis of the 16S rRNA gene was performed and 100% identity was found with different strains of A. platys. These data confirm that cats are susceptible to parasitism by A. platys.

Potencial acaricida do óleo de andiroba Carapa guianensis Aubl. sobre fêmeas adultas ingurgitadas de Anocentor nitens Neumann, 1897 e Rhipicephalus sanguineus Latreille, 1806

Avaliou-se o potencial acaricida in vitro do oleo da semente da andiroba (Carapa guianensis) sobre femeas ingurgitadas de Anocentor nitens (n=210) e Rhipicephalus sanguineus (n=140), coletadas manualmente, respectivamente, de equinos e de caes naturalmente infestados. Para o teste de imersao, empregaram-se cinco diluicoes do oleo de andiroba, 100%, 50%, 30%, 25% e 10%, em agua destilada, utilizando-se tween 80 como dispersante. No teste com A. nitens, foram usadas tres repeticoes para cada diluicao, utilizando-se 10 femeas ingurgitadas para cada tratamento. No teste com R. sanguineus, usaram-se duas repeticoes, e formaram-se, ainda, dois grupos-controle para cada especie de ixodideo, um com agua destilada e outro com tween 80 mais agua destilada. Apos os testes, as femeas foram mantidas em laboratorio sob temperatura ambiente. Observou-se mortalidade das femeas ingurgitadas e reducao de postura, neste caso, com ovos inferteis, demonstrando eficacia de 100% nas duas especies em todas as diluicoes testadas. Os dados obtidos evidenciaram a potencialidade do uso do extrato de andiroba contra A. nitens e R. sanguineus.

Comparação de nested-PCR com o diagnóstico direto na detecção de Ehrlichia canis e Anaplasma platys em cães

The clinical signs of Ehrlichia canis and Anaplasma platys infection are similar, and the diagnosis of these pathogens made by stained blood smears is poor due sensibility and specificity. On the other hand, the molecular diagnosis is highly sensitive and specific and nested-PCR have been optimized for accurate diagnosis these pathogens in dogs. At the veterinary teaching hospital, whole-blood samples with EDTA were obtained from 100 dogs and smears were made from blood samples for evaluation for intracellular parasites. For each sample, DNA was extracted and submitted to nPCR analysis for detection of E. canis and A. platys. The results of stained blood smears showed 9% of the animals were positive for E. canis and 21% for A. platys. Regarding of nPCR analysis, 57 and 55% of dogs were positive for E. canis and A. platys respectively. As compared to a nested PCR, the stained blood smears revealed false-negative results for both E. canis and A. platys. The results indicate that the nPCR is highly sensitive and specific for detection of both pathogens and the molecular diagnosis could be more useful at veterinary hospital.

First report and molecular characterization of Trypanosoma vivax in cattle from state of Pernambuco, Brazil

This paper describes an outbreak of Trypanosoma vivax for the first time in the state of Pernambuco, Brazil, affecting dairy cattle in the municipality of Itambé in the northern coastal zone of the state. Clinical signs compatible with infection by blood protozoa and epidemic miscarriages were observed. The diagnosis of T. vivax was confirmed through biometric microscopy and molecular analysis with PCR and DNA sequencing. The T. vivax isolate detected in the present study proved to be genetically very close to other Brazilian isolates of the protozoan despite being geographically distant.

Epidemiologic surveillance of canine visceral leishmaniasis in the municipality of Recife, Pernambuco

This paper describes an epidemiologic investigation carried out in Imbiribeira district, where a canine visceral leishmaniasis case was recorded. Despite the absence of seropositive dogs and sand flies, these findings are not sufficient to discard the occurrence of a zoonotic cycle of visceral leishmaniasis in Recife.

Polymerase chain reaction and real-time PCR for diagnosing of Leishmania infantum chagasi in dogs

The importance of dogs as a reservoir for Leishmania infantumchagasi in urban environments has stimulated numerous studies assessing diagnostic techniques. When performed properly, such procedures are an important step in preventing leishmaniasis in humans. Molecular methods have become prominent for this purpose. The aim of the present study was to determine the performance of the polymerase chain reaction (PCR) and real-time PCR (qPCR) for diagnosing of canine visceral leishmaniasis (CVL) using different biological samples. For this, 35 dogs from an area endemic for CVL were used. Bone marrow aspirate and lymph node and spleen fragments from these dogs were used for the molecular diagnosis. In the present study, qPCR was able to detect a greater number of positive animals than seen with PCR. Among the different biological samples used, there was no significant difference in L. infantumchagasi DNA detection between PCR and qPCR. However, considering that lymph nodes are easy to acquire, these can be considered to be the best samples for making molecular diagnoses of L. infantum chagasi infection.

Serological survey of Neospora caninum in small ruminants from Pernambuco State, Brazil

Neospora caninum infection is an important cause of bovine abortion as well as neonatal mortality in goats and sheep. A serological survey for antibodies against N. caninum in goats and sheep was carried out in the municipality of Ibimirim, PE, Northeastern Brazil. The imunnofluoresce antibody test showed that 26.6% (85/319) of the goats and 64.2% (52/81) of the sheep were positive. Serologic reactivity was associated with age in goats (p < 0.01) and sheep (p > 0.05), with increasing rates in older animals. These results indicate exposure to N. caninum among small ruminants in the study area.

Occurrence of Cryptosporidium spp. in Antillean Manatees (Trichechus manatus) and Amazonian Manatees (Trichechus inunguis) from Brazil

Abstract: Infections by Cryptosporidium spp. in aquatic mammals is a major concern due to the possibility of the waterborne transmission of oocysts. The aim of the present study was to report the occurrence of Cryptosporidium spp. in Antillean manatees (Trichechus manatus) and Amazonian manatees (Trichechus inunguis) from Brazil. Fecal samples were collected and processed using Kinyouns method. Positive samples were also submitted to the direct immunofluorescence test. The results revealed the presence of Cryptosporidium spp. oocysts in 12.5% (17/136) of the material obtained from the Antillean manatees and in 4.3% (05/115) of the samples from the Amazonian manatees. Cryptosporidium spp. infection was more prevalent in captive animals than in free-ranging specimens.