Epidemiological aspects of equine babesioses in a herd of horses in Brazil.

Epidemiological studies of Babesia equi and B. caballi were undertaken in a herd of 120 pastured horses in Rio de Janeiro, Brazil. The area where the horses were held was shown to be highly endemic for both Babesia spp., i.e. the prevalence of B. equi antibodies in horses aged 6 months or older ranged from 90.6% to 100% as determined by the immunofluorescence antibody (IFA) test, and the prevalence of B. caballi antibodies as determined by Western blot ranged from 59.4% to 65.5%. From the herd, 20 foals and their dams were selected to estimate the degree of tick infestation and the foals were bled at monthly intervals to determine the incidence of antibodies to B. equi and B. caballi. The incidence of B. equi was 100% by about 127 days of age as determined by IFA of B. caballi was 100% by about 150 days of age as determined by Western blot. Tick infestation of the horses estimated by using a semiquantitative key ranged from at least five ticks on every horse to more than 100 ticks on many horses throughout the year. Except for three Boophilus microplus female ticks, they were identified as Amblyomma cajennense and Anocentor nitens. A. cajennense had one generation per year, whereas An. nitens had three. Kinetes of B. caballi were detected in the haemolymph of two of 68 An. nitens female ticks and in the ovary and eggs of one of these, suggesting that this tick is a significant vector of B. caballi.(ABSTRACT TRUNCATED AT 250 WORDS)

Diagnosis of Babesia caballi infections in horses by enzyme-linked immunosorbent assay (ELISA) and western blot.

From Babesia caballi in vitro cultures a preparation of 100% infected erythrocytes was obtained. From this, B. caballi antigens were extracted with the detergent 3-[(3-Cholamidopropyl)-dimethylammonio]-1-propane-sulfonate (CHAPS) and used as ELISA antigens. A control antigen of normal erythrocytes from the same donor horse was prepared in an identical manner. The ELISA and Western blot were validated by testing of sera from horses experimentally infected with B. caballi or B. equi or not infected with Babesia spp. ELISA and Western blot results were compared with those obtained by the immunofluorescence antibody test (IFAT) and complement fixation test (CFT). The sensitivity of the ELISA of 98.3% obtained for sera from day 14 after infection was superior to the Western blot (94.9%), the IFAT (96.6%) and the CFT (28.8%). No positive results were obtained in the ELISA and Western blot with 106 sera from horses not infected with Babesia spp. resulting in a calculated specificity of 100% for both tests. Cross reactions of B. equi-positive sera did occur to a larger extent in the ELISA (20%) than in the IFAT (4%). No cross reactions were observed with the Western blot and the CFT. The higher sensitivity of the ELISA was also demonstrated by testing of 132 field sera: more positive results were obtained by ELISA (112) as compared to IFAT (92) or CFT (41). The validity of these results was confirmed by testing of sera by Western blot. The ELISA as the most sensitive test provides the best method for the identification of carrier horses to prevent the introduction into non-endemic areas (export testing).(ABSTRACT TRUNCATED AT 250 WORDS)

Identification of diagnostic antigens for South American Babesia caballi infections.

Sera from 60 horses held in breeding herd in Brazil were examined monthly by ELISA, immunofluorescence antibody test (IFAT) and Western blot. All foals had maternal antibodies detectable by ELISA and IFAT, and sero-conversion took place between the 2nd and 5th month of age. The 48 and 50 kDa antigens were recognized first in the course of infection. Of 79 sera taken after sero-conversion 78 reacted with the 48 kDa antigen, 76 with the 50 kDa, 50 with the 70 kDa, 54 with the 112 kDa, 72 with the 141 kDa antigen. In general, sera from horses older than 1 year reacted with all 5 diagnostic antigen bands, but sera from horses older than 3 years only weakly. The antigens of 48 and 50 kDa appear to be conserved among all strains of B. caballi examined so far and are consistently recognized by all infected horses. They are the target antigens for a serological test based on antigens produced by recombinant DNA techniques.