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ABSTRACT: The objective of this study was to design and evaluate new primers for species-specific detection of L. infantum chagasi using PCR. Two combinations of primer pairs were established with the aim of obtaining specific amplification products from the L. infantum chagasi 18S rRNA gene. The combinations of the primer pairs and the respective sizes of the PCR products, based on the U422465 GenBank reference sequence of L. infantum chagasi, were: LCS1/LCS3 (259 bp) and LCS2/LCS3 (820 bp). It was concluded that the new PCR assays optimized using the primer pairs LCS1/LCS3 and LCS2/LCS3 were effective for specific detection of L. infantum chagasi, with analytical sensitivity to detect 1 pg/µL of DNA.
Revista brasileira de parasitologia veterinaria = Brazilian journal of veterinary parasitology: Orgao Oficial do Colegio Brasileiro de Parasitologia Veterinaria 09/2012; 21(3):304-7. · 0.46 Impact Factor
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ABSTRACT: The genus Babesia comprises protozoa that cause diseases known as babesiosis. Dogs are commonly affected by Babesia canis or Babesia gibsoni. Babesia canis is divided into the subspecies Babesia canis canis, Babesia canis vogeli and Babesia canis rossi. Among these, Babesia canis vogeli predominates in Brazil. The objective of this study was to conduct a phylogenetic analysis on Babesia isolates from dogs in Goiânia, Goiás. Blood samples were obtained from 890 dogs presenting clinical signs suggestive of canine babesiosis that were attended at a veterinary hospital of Goiás. Only samples presenting typical intraerythrocytic parasites were used in the study. These were subjected to DNA extraction and amplification of a fragment of the 18S rRNA, by means of PCR. The PCR products were purified and sequenced. Sequences were obtained from 35 samples but only 17 of these were kept after quality assessment. Similarity analysis using BLASTn demonstrated that all 17 sequences corresponded to B. canis vogeli. Analysis using the Mega4 software showed that the isolates of B. canis vogeli from dogs in Goiânia present a high degree of molecular similarity (99.2 to 100%) in comparison with other reference isolates from other regions of Brazil and worldwide, deposited in GenBank.
Revista brasileira de parasitologia veterinaria = Brazilian journal of veterinary parasitology: Orgao Oficial do Colegio Brasileiro de Parasitologia Veterinaria 10/2011; 20(4):274-80. · 0.46 Impact Factor
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ABSTRACT: Canine babesiosis is an infectious disease caused by either Babesia gibsoni or Babesia canis protozoans. The latter is also classified under three different phylogenetic groups, referred to as subspecies B. canis canis, B. canis vogeli and B. canis rossi. The objective of the present study was to validate and standardize a PCR assay to discriminate the organisms at the subspecies level. First, the reference sequences of the 18S rRNA, 5.8S rRNA and 28S rRNA genes, including the internal transcribed spacer 1 (ITS1) and 2 (ITS2) of the most common species and subspecies of the genus Babesia were retrieved from the GenBank database. Subspecies-specific primers (BAB3, BAB4 and BAB5) and one genus-specific primer were designed from the alignment of the sequences. The PCR assays were evaluated in three different combinations of primer pairs in order to assure complete specificity for each reaction. The results of the tests had demonstrated effectiveness of the novel primer pairs BAB1/BAB3, BAB1/BAB4 and BAB1/BAB5 for the amplification of the subspecies-specific target fragments of 746 bp (B. c. canis), 546 bp (B. c. vogeli) and 342 bp (B. c. rossi) by PCR. The original enzymatic amplification assays with novel primers reported in this paper were confirmed to be a reliable tool for the specific discrimination among B. canis subspecies by single-step PCR assays.
Veterinary Parasitology 04/2008; 152(1-2):16-20. · 2.58 Impact Factor
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ABSTRACT: O trabalho foi realizado com o objetivo principal de obter isolados autóctones puros de Anaplasma marginale a partir de amostra de sangue de bovino criado em uma área endêmica para a tristeza parasitária bovina no município de Goiânia. Obteve-se o inóculo para o isolamento de A. marginale de animal doador, portador de infecções naturais mistas, após receber tratamento seletivo com dose babesicida esterelizante de dipropionato de imidocarb. Dois dias após esse tratamento, colheu-se dele uma amostra de sangue de 20 mL do animal doador para inoculação em bezerro neonato privado de colostro e livre de infecções por hemoparasitos. Após trinta dias da inoculação, o receptor apresentou febre, apatia e parasitemia patente por A. marginale. Amostras de sangue foram então colhidas e preparadas na forma de estabilizados para serem criopreservadas e, assim, comporem banco de isolados de hemoparasitos autóctones. Realizou-se a comprovação da pureza do isolado, concomitantemente, pela demonstração de soroconversão específica, pela reação de PCR e, ainda, pela subinoculação da amostra criopreservada em bezerro suscetível. Constatou-se ainda que o uso de bezerros neonatos privados de colostro, como animais suscetíveis, pode ser considerado como modelo prático, eficaz e relativamente barato para o isolamento de hemoparasitos em regiões endêmicas. PALAVRAS-CHAVES: Anaplasma marginale, anaplasmose bovina, bezerros, isolados. This study was conducted with the objective of obtaining pure autochthonous isolates of Anaplasma marginale from blood samples of bovines that have been raised in the tick-borne disease endemic area of the municipality of Goiânia. The inoculum of A. marginale was prepared from a donor animal, known to be natural carrier of hemoparasite mixed infections, after treatment with a babesicid sterilizing dose of imidocarb dipropionate. A 20 mL blood sample was collected from the donor animal 2 days post-infection and then transferred to a colostrum-deprived and hemoparasite free newborn calf. The receptor animal showed signs of fever and apathy with patent A. marginale parasitemia 30 days after inoculation. Blood sample was then collected and prepared as stabilates to be cryopreserved and to take part of an autochthonous blood parasites isolates library. The isolate purity was assured concomitantly by specific seroconversion tests, by PCR and by finally by the subinoculation of susceptible calves with the cryopreseved stabilates. It was confirmed that the use of colostrum-deprived free newborn calves as susceptible animals may be considered a practical and effective model to obtain pure hemoparasite isolates in endemic areas with additional advantages such as feasibility and low cost. Key words: Anaplasma marginale, isolates, calves, bovine anaplasmosis.
Ciência Animal Brasileira. 01/2008;
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ABSTRACT: Studies on prevalence of anti-Toxoplasma gondii antibodies was carried out by the indirect hemaglutination test in 109 caprine sera samples from five distinct heards in Goiânia, Goiás. It had been shown positivity of 30.2% for animals under 1 year old and 51.5% for animals over 1 year old. Between females 46.3% were positive and males 33.3%. And finally 43.1% among animals of both sexes and any age, except lactents. The sera were tested from the inicial dilutron of 1/64.
Pesquisa Agropecuária Tropical. 01/2007;
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ABSTRACT: The bovine tripanosomiasis by Trypanosoma vivax was reported for the first time in Brazil by Shaw & Lainson, in 1972, in Pará. Recently it has been reported in several areas of the Pantanal Mato-Grossense Region. The objective of the present paper was to report, for the first time, the occurrence of T. vivax in the State of Tocantins, in a Brahman herd of 250 animals, which had been introduced into a property in the municipality of Formoso do Araguaia, coming from São Paulo. From animals presenting signs of weakness, 9 was randomly chosen for detailed clinical assessment and laboratory exams such as blood smears and hematocrit. These animals presented weigh loss, edema, fever and mucous membrane paleness. The microscopic smear examination revealed high parasitemias for T. vivax in samples of three animals, with the packet cell volumes ranging between 15 and 20%. The herd history, the ecosystem of the area and the results of the clinical and laboratory exams confirmed the occurrence of an outbreak of trypanosomiasis by T. vivax in a bovine herd in the State of Tocantins.
KEY WORDS: Trypanosoma vivax, bovine trypanosomiasis, Tocantins, epidemiology. A tripanossomíase bovina por Trypanosoma vivax foi registrada pela primeira vez no Brasil por SHAW & LAINSON, em 1972, no Pará. Recentemente, tem sido reportada em várias regiões do Pantanal Mato-Grossense. O presente trabalho teve como objetivo relatar, pela primeira vez, a ocorrência de T. vivax no Estado do Tocantins, em um rebanho composto de 250 animais da raça Brahman, recém-introduzido em uma propriedade no município de Formoso do Araguaia, procedente de São Paulo. Esc olheram- se ao acaso nove animais que apresentavam sinais de debilidade para a execução de exames clínicos detalhados e colheita de sangue para preparo de esfregaços sangüíneos e determinação do hematócrito. Entre os animais examinados observaram-see emagrecimento, edema de barbela, febre e palidez de mucosa. O exame microscópico revelou parasitemias elevadas por T. vivax em amostras de três animais, cujos hematócritos apresentavam valores entre 15% e 20%. O histórico do rebanho, o ecossistema da região e os resultados dos exames clínicos e laboratoriais confirmaram a ocorrência de um surto de tripanossomíase por T. vivax em um rebanho no Estado do Tocantins.
PALAVRAS-CHAVE: Epidemiologia, Trypanosoma vivax, Tocantins, tripanossomíase bovina.
Ciência Animal Brasileira. 01/2006;
