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      An enzyme-linked immunosorbent assay for the detection of IgG antibodies against Babesia equi in horses Translated title: Um ensaio imunoenzimático para a detecção de anticorpos IgG contra Babesia equi em eqüinos

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          Abstract

          A crude antigenic preparation of Babesia equi was used to develop and establish the suitability of an enzyme-linked immunosorbent assay (ELISA) for the detection of parasite carriers. Optimal dilutions of the antigen, using positive and negative reference sera, were determined by checkboard titrations. The specificity and sensitivity of the ELISA were 100 %. A total of 90 serum samples were taken from horses from the Northeast region of São Paulo State and examined for diagnosis of equine B. equi infection by ELISA. Approximately 75% (n=67) of all the horses tested were found serologically positive for B. equi. These results suggest that the ELISA described may prove to be an appropriate serological test for epidemiological studies on B. equi infections in the field and that equine piroplasmosis is a cause for serious concern in the State of São Paulo, Brazil.

          Translated abstract

          Um ensaio de imunoadsorção enzimática (ELISA) baseado em antígeno bruto de Babesia equi foi desenvolvido para a detecção de portadores crônicos de babesiose eqüina. As diluições ótimas do antígeno e dos soros controles positivo e negativo foram determinadas através de titulação em bloco. A sensibilidade e especificidade do teste foram de 100%. O ELISA foi empregado em 90 soros de eqüinos da região Nordeste do Estado de São Paulo, de modo que aproximadamente 75% (n=67) dos eqüinos foram positivos para B. equi. Estes resultados sugerem que o ELISA descrito pode ser utilizado no diagnóstico sorológico de B. equi e que a piroplasmose eqüina é um problema sério no 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 an update on diagnosis, treatment and prevention.

            A Brüning (1996)
            Two haemoprotozoan parasites, Babesia caballi and Babesia equi, can cause equine piroplasmosis. Due to the presence of potential tick vectors in areas so far unaffected by equine babesias, import and export regulations often require the serum testing of animals for evidence of infection. Although the complement fixation test (CFT) has been recommended for detecting the presence of antibodies to Babesia spp., it has been demonstrated to have several disadvantages, including false-positive results and low sensitivity for detecting latent infections. An enzyme-linked immunosorbent assay (ELISA) may be an alternative for increased and sensitive detection of acute and latent babesial infections, but its development to date has been hindered by a limited antigen supply and poor specificity. In vitro cultivation of both parasite species and the identification of parasite proteins for diagnostic use has facilitated the development of a highly sensitive and specific ELISA. For the direct detection of the parasites, DNA probes are now available. Several drugs are available for the treatment of equine piroplasmosis. For instance, diminazene diaceturate is effective in the chemosterilization of B. caballi and in the elimination of clinical signs in B. equi infections. Antitheilericidal drugs such as buparvaquone have been demonstrated to be effective in combatting disease due to B. equi and may--in combination with imidocarb--also eliminate the parasite. The control of equine piroplasmosis must include effective tick control, seromonitoring of animals and the application of chemotherapy.
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              An enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies against Babesia bovis in cattle.

              A method for the isolation of Babesia bovis merozoites from infected erythrocytes (Machado et al., 1994) and an enzyme-linked immunosorbent assay (ELISA) for the detection of anti-B. bovis antibodies were developed. This ELISA utilizes a soluble, alkali-digested B. bovis antigen. Sera from calves experimentally infected with B. bovis were screened by this technique from day 9 to day 233 postinfection (PI). Maximum titers were reached between days 29 and 149 PI. Sera from calves (n = 62), heifers (n = 38) and cows (n = 49), raised in tick-infested areas of São Paulo State, showed higher antibody levels in heifers and cows. A higher percentage of negative sera (19.4%) was found among calves. Sodium dodecyl sulphate-polyacrylamide electrophoresis (SDS-PAGE) and immunoblotting have identified proteins of similar molecular mass in the two species. Sera from calves experimentally infected with B. bovis reacted with homologous antigens at the level of 95, 66 and 23 kDa. The same serum reacted with the 23 kDa band of heterologous antigen. Sera from calves experimentally infected with B. bigemina recognized 82, 66, 58, 36 and the 23 kDa polypeptides of homologous and heterologous antigens. The experimental ELISA described may prove to be a practical serological test for bovine Babesia infection with the choice of specific test antigen for B. bovis and B. bigemina.
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                Author and article information

                Journal
                cr
                Ciência Rural
                Cienc. Rural
                Universidade Federal de Santa Maria (Santa Maria, RS, Brazil )
                0103-8478
                1678-4596
                October 2004
                : 34
                : 5
                : 1525-1529
                Affiliations
                [03] orgnameUniversidade Federal Rural do Rio de Janeiro orgdiv1Departamento de Clínica e Cirurgia
                [02] Jaboticabal SP orgnameUniversidade Estadual Paulista orgdiv1Faculdade de Ciências Agrárias e Veterinária orgdiv2Departamento de Patologia Animal zacarias@ 123456fcav.unesp.br
                [04] orgnameUniversidade Federal Rural do Rio de Janeiro orgdiv1Departamento de Parasitologia Animal
                [01] orgnameUniversidade Estadual Paulista orgdiv1Faculdade de Ciências Agrárias e Veterinária
                Article
                S0103-84782004000500031 S0103-8478(04)03400531
                33a4f1ed-158c-4709-9910-2f20f39ffa50

                This work is licensed under a Creative Commons Attribution 4.0 International License.

                History
                : 26 September 2003
                : 28 January 2004
                Page count
                Figures: 0, Tables: 0, Equations: 0, References: 17, Pages: 5
                Product

                SciELO Brazil

                Categories
                Parasitology

                Parasitology
                Babesia equi,piroplasmose eqüina,ELISA,equine piroplasmosis
                Parasitology
                Babesia equi, piroplasmose eqüina, ELISA, equine piroplasmosis

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