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      Occurrence of Theileria equi in horses raised in the Jaboticabal microregion, São Paulo State, Brazil Translated title: Ocorrência de Theileria equi em equinos criados na microrregião de Jaboticabal, Estado de São Paulo, Brasil

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          Abstract

          Blood and serum samples from 170 horses raised in the Jaboticabal microregion, São Paulo State, Brazil, were collected and tested by microscopic examination of blood smears, indirect fluorescent antibody test (IFAT) and nested polymerase chain reaction (nPCR) for Theileria equi infections. The association among the test results was verified by the McNemar test. During the examination of thin blood smears, parasites were detected in six (3.52%) horses. Anti-T. equi antibodies were detected in 100% sera samples, with titers ranging between 1:80 and 1:5120. The nPCR based on the T. equi merozoite antigen gene (EMA-1) allowed the visualization of specie-specific amplified product in 108 (63.53%) horses. All six samples judged positive microscopically were also positive for nPCR. Statistical analysis indicated general disagreement (p < 0.0001) between IFAT and nPCR; IFAT and blood smear; and nPCR and blood smear on the detection of parasite carriers. The results of the present study indicate that T. equi is widely spread among horses in the Jaboticabal microregion, Northeast region of São Paulo State, Brazil.

          Translated abstract

          Amostras de sangue e soro de 170 equinos criados na microrregião de Jaboticabal, Estado de São Paulo, Brasil, foram coletadas e avaliadas pelo exame direto em esfregaço sanguíneo, reação de imunofluorescência indireta (RIFI) e nested reação em cadeia da polimerase (nPCR) para a detecção de infecções por Theileria equi. A concordância dos resultados entre os testes de diagnóstico foi verificada pelo teste de McNemar. Durante o exame dos esfregaços sanguíneos, parasitos foram detectados em seis (3,52%) equinos. Anticorpos anti-T. equi foram detectados em 100% das amostras de soros, com títulos variando entre 1:80 e 1:5120. O nPCR, baseado na sequência do gene do antígeno de merozoíto de T. equi (EMA-1), permitiu a visualização de produtos de amplificação espécie-específico em 108 (63,53%) equinos. Houve diferença altamente significativa (p < 0,0001) entre RIFI e nPCR; RIFI e esfregaço sanguíneo; e nPCR e esfregaço na detecção do parasito. O resultado do presente estudo indica que a infecção por T. equi está amplamente distribuída entre os equinos na microrregião de Jaboticabal, região Nordeste do Estado de São Paulo, Brasil.

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          Current state and future trends in the diagnosis of babesiosis.

          An overview is given of the currently available methods to diagnose babesiosis in livestock. Microscopic techniques are still the only appropriate techniques to diagnose acute disease. Thin or thick blood films stained with Giemsa's stain are sufficient. The sensitivity ranges from 10(-5) to 10(-6), i.e. one parasite per 10(5)-10(6) erythrocytes can be detected. Thick films stained with acridine orange (sensitivity approximately 10(-7)) and the Quantitative Buffy Coat (QBC) analysis tube system (sensitivity approximately 10(-7)-10(-8)) are applicable for diagnosis in the laboratory. DNA probes are very specific tools to identify haemoparasites in organs post mortem and in ticks. For the identification of carrier animals the sensitivity (approximately 10(-5)-10(-6)) is generally not sufficient. For the latter the polymerase chain reaction (PCR) technique is a very powerful tool (sensitivity approximately 10(-9)). Many different serodiagnostic tests have been described; however, the immunofluorescence antibody test is the most widely used, while the enzyme-linked immunosorbent assay (ELISA) is the test system which holds the greatest promise for the future. Thus far, improvements to the ELISA have been limited as the quality of antigen preparations made from infected blood is generally poor with a few exceptions (Babesia bovis, Babesia caballi). Potentially, most of the problems associated with crude antigens can be overcome by the production of recombinant antigens. Several ELISAs based on highly defined recombinant antigens have been described and show promise. None of these tests has been validated to the extent that it could be applied globally. Future research requirements as well as the need for coordination of the research effort and collaboration between institutions involved in the diagnosis of babesiosis are discussed.
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            Equine piroplasmosis: a review.

            This review focuses on equine piroplasmosis with specific reference to its distribution, diagnosis and clinical and pathological signs. The more common used drugs are discussed both with reference to treatment and chemosterilization. Areas requiring further research are also briefly mentioned.
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              Sequence heterogeneity in the 18S rRNA gene within Theileria equi and Babesia caballi from horses in South Africa.

              A molecular epidemiological survey of the protozoal parasites that cause equine piroplasmosis was conducted using samples collected from horses and zebra from different geographical locations in South Africa. A total of 488 samples were tested for the presence of Theileria equi and/or Babesia caballi using the reverse line blot hybridization assay. Ten percent of the samples hybridized to the Theileria/Babesia genus-specific probe and not to the B. caballi or T. equi species-specific probes, suggesting the presence of a novel species or genotype. The small subunit of rRNA gene (18S; approximately 1600bp) was amplified and sequenced from 33 of these 488 samples. Sequences were compared with published sequences from the public sequence databases. Twelve distinct T. equi and six B. caballi 18S rRNA sequences were identified. Alignments demonstrated extensive sequence variation in the V4 hypervariable region of the 18S rRNA gene within T. equi. Sequence variation was also found in B. caballi 18S rRNA genes, although there was less variation than observed for T. equi. Phylogenetic analysis based on 18S rRNA gene sequences revealed three T. equi clades and two B. caballi clades in South Africa. The extent of sequence heterogeneity detected within T. equi and B. caballi 18S rRNA genes was unexpected since concerted evolution is thought to maintain homogeneity within repeated gene families, including rRNA genes, in eukaryotes. The findings reported here show that careful examination of variants of the 18S rRNA gene of T. equi and B. caballi is required prior to the development of molecular diagnostic tests to detect these parasites in horses. Species-specific probes must be in designed in regions of the gene that are both conserved within and unique to each species.
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                Author and article information

                Contributors
                Role: ND
                Role: ND
                Role: ND
                Journal
                rbpv
                Revista Brasileira de Parasitologia Veterinária
                Rev. Bras. Parasitol. Vet.
                Colégio Brasileiro de Parasitologia Veterinária (Jaboticabal )
                1984-2961
                December 2010
                : 19
                : 4
                : 228-232
                Affiliations
                [1 ] Universidade Federal Rural do Rio de Janeiro Brazil
                [2 ] Universidade Estadual Paulista Brazil
                Article
                S1984-29612010000400007
                10.1590/S1984-29612010000400007
                c3abf022-055c-41ac-8a38-baada0808a8c

                http://creativecommons.org/licenses/by/4.0/

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                SciELO Brazil

                Self URI (journal page): http://www.scielo.br/scielo.php?script=sci_serial&pid=1984-2961&lng=en
                Categories
                PARASITOLOGY
                VETERINARY SCIENCES

                Parasitology,General veterinary medicine
                Theileria equi,horse,diagnosis,IFAT,nested PCR,equino,diagnóstico,RIFI
                Parasitology, General veterinary medicine
                Theileria equi, horse, diagnosis, IFAT, nested PCR, equino, diagnóstico, RIFI

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