Detection and molecular characterization of Babesia caballi and Theileria equi isolates from endemic areas of Brazil.

Blood samples were collected from 487 adult horses, including 83 pregnant mares, at a slaughterhouse located in Araguari, Minas Gerais State, Brazil. For each blood sample, the packed cell volume (PCV) was determined, and Giemsa-stained smears were microscopically examined for the presence of hemoparasites. The plasma was examined by the indirect fluorescent antibody test for detection of antibodies against Babesia caballi and Theileria equi. In addition, DNA was extracted and analyzed by a multiplex real-time polymerase chain reaction (PCR), specific for B. caballi and T. equi. Products of PCR were sequenced and compared with each other and with known sequences. The serological results showed a total prevalence of 91.0% for T. equi and 83.0% for B. caballi, while by PCR, prevalences of 59.7% for T. equi and 12.5% for B. caballi were observed. However, no correlations were seen between positivity (neither by serology nor by PCR) and PCV values. As expected, the microscopic examination of blood smears showed low sensitivity in detecting the infections when compared to the PCR. Only 35 out of 570 blood smears were positive, with parasitemias below 0.1%. No congenital transmission was detectable. As far as sequencing is concerned, no differences were seen among the isolates of each species nor among them and known sequences available. These results confirm, by molecular methods, the high prevalence rates of T. equi and B. caballi infections in carrier horses in Brazil. However, no diversity was observed among the isolates within the studied regions.

Application of Taenia saginata metacestodes as an alternative antigen for the serological diagnosis of human neurocysticercosis.

Serological tests are an important tool for the diagnosis of neurocysticercosis (NCC), the disease caused by Taenia solium metacestodes. The aim of the present research was to test the application of Taenia saginata metacestodes as an alternative antigen for use in the immunofluorescence antibody test (IFAT), enzyme-linked immunosorbent assay (ELISA), and Western blotting (WB) tests compared with the metacestodes antigen of T. solium in serum samples. The samples were obtained from 130 individuals: 20 from patients with definitive NCC, Group 1; 18 from individuals infected by Taenia sp., Group 2; 40 from individuals infected by various parasites, Group 3; and 40 from healthy individuals, Group 4. The sensitivity of IFAT, ELISA, and WB using antigen obtained from T. solium applied to the patients of Group 1 yielded results of 85, 95, and 95%, respectively, for the three tests. When the tests were conducted using T. saginata metacestodes, results were 75, 80, and 85%, respectively. The specificity of IFAT, ELISA, and WB using antigen obtained from T. solium yielded results of 94.9, 88.8, and 93.9%. When the tests were conducted using T. saginata metacestodes, results were 95.9, 88.8, and 93.6%, respectively. No statistical differences for sensitivity or specificity among the antigens were found. In conclusion, the results indicated that T. saginata metacestodes can be used as an alternative antigen for NCC diagnosis.

Seroprevalence of Toxoplasma gondii infection in goats by the indirect haemagglutination, immunofluorescence and immunoenzymatic tests in the region of Uberlândia, Brazil

A comparative study of the indirect haemagglutination (IHA), immunofluorescence (IFAT) and immunoenzymatic (ELISA) tests was carried out to determine the prevalence of Toxoplasma gondii antibodies in goats. One hundred seventy-four serum samples were obtained from four goat herds from the region of Uberlândia, State of Minas Gerais. The distribution of the animals, according to their origin, was as follow: 71 from herd I; 39 from herd II; 37 from herd III; and 27 from herd IV. Serum samples were analyzed by IHA, IFAT and ELISA, considering the reactivity of the serum samples at dilution ≥ 1:64 as cut off titer for the three tests. A global seroprevalence of 18.4 percent was observed, with significantly higher positivity rate in the herd II (66.7 percent) and older animals (> 36 months). A high and significant positive correlation was found between the titers obtained by the IHA versus IFAT, IHA versus ELISA, and ELISA versus IFAT. Therefore, it can be concluded that the three analyzed tests have shown to be highly concordant and appropriate for epidemiological surveys of Toxoplasma infection in goats. Although the seroprevalence of T. gondii infection in goats is relatively low in this region as compared to other regions of the country, adequate management might be useful and essential to control the infection in the goat herds

Detection of Toxoplasma gondii specific antibodies in dogs: a comparative study of immunoenzymatic, immunofluorescent and haemagglutination titers

We evaluated the titers os anti-T. gondii antibodies by various serological testes in 40 serum samples from dogs exhibiting clinical signs of infectios diseases. Indirect immunoflurescence (IgG-IFI), indirect haemagglutination (IHA and M-Toxo) and immunoenzymatic (ELISA and PA-ELISA) tests were carried out. Titers > 64 were considered as positive. Anti-Toxoplasma antibodies were found in 9 (22.5 per cent), 14 (35 per cent) and 12 (30 per cent) samples, respectively for IHA, IgG-IFI, ELISA and PA-ELISA. The results showed that 57 per cent were negative in all tests and 43 per cent of the dogs presented antibodies to T. gondii; from these, 20 per cent were positive in all three tests with high titers of antibodies and 23 per cent were positive in only one or two tests with low titers of antibodies and mainly related to the IFI and ELISA tests. We observed 5 (12.5 per cent) and 1 (2.5 per cent) reactive samples, respectively, by M-Toxo and IHA with or wihout 2-mercapthoethanol, in the attempt to detect specific IgM. We can conclude that serodiagnosis of toxoplasmosis in dog have to be based on the combination of serological tests (IFI and ELISA) and with emphasis at the determination of the titers and the classes of specific antibodies.