Investigations on naturally occurring Trypanosoma evansi infections in horses, cattle, dogs and capybaras (Hydrochaeris hydrochaeris) in Pantanal de Poconé (Mato Grosso, Brazil).

The prevalence of Mal de Cadeiras--Portuguese for Trypanosoma (T.) evansi infections in horses--as well as the prevalence of T.evansi infections in cattle, dogs and free-ranging capybaras (Hydrochaeris hydrochaeris) was investigated in Pantanal de Poconé (Mato Grosso, Brazil). In 0.3, 8.6 and 8.0% of the horses, dogs and capybaras, respectively, infection was detected using standard parasitological methods. A seroprevalence of 4.1, 2.3, 7.1 and 22.0% was found in horses, cattle, dogs and capybaras, respectively, using an enzyme-linked immunosorbent assay for the detection of T.evansi antigen (Ag-ELISA), whereas 9.6, 4.2, 18.6 and 14.0% of the animals investigated were reactive in a T.evansi antibody (Ab-) ELISA. Positive ELISA results ('high responders') were identified using computer-assisted mixture analysis (C.A.MAN). Agglutinating antibodies were detected by the T.evansi card agglutination test for trypanosomiasis (CATT/T.evansi) in 14.6%, 1.3%, 15.7% and 22.0% of the horses, cattle, dogs and capybaras, respectively. A moderate but significant (kappa test; p < 0.05) agreement beyond chance level was observed between Ab-ELISA and CATT results but generally not between antibody and antigen detection tests. Therefore, in an attempt to maximize the information yield of the three serodiagnostic tests, their results were numerically scored (negative = 0, intermediate = 1, positive = 2) and added up to a total score (TS) which was considered indicative for infection when TS > or = 4 (results of the Ag-ELISA received double weight). Estimates of seroprevalence according to TS were 13.2, 4.7, 30.0 and 24.0% for horses, cattle, dogs and capybaras, respectively. Identical isoenzyme profiles, known as zymodeme 58 (T.evansi MCAN/BR/86/H), were found in all T.evansi stocks isolated in the study area (six from dogs, one from a horse and one from a capybara). From the results of this study it can be inferred that Mal de Cadeiras is endemic in Pantanal de Poconé. Although clinical and parasitological findings support the possible role of the capybara as a reservoir host of T.evansi, dogs and cattle--due to their close contact with horses--should rather be regarded as efficient reservoir hosts for Mal de Cadeiras in the study area.

The use of DNA hybridization and numerical taxonomy in determining relationships between Trypanosoma brucei stocks and subspecies.

The nuclear DNAs of 71 trypanosome stocks from different African countries, representative of the three Trypanosoma brucei subspecies, and one T. evansi stock, have been analysed by the combined use of restriction endonuclease digestion, gel electrophoresis and molecular hybridization with both trypanosome surface-antigen-specific and undefined genomic DNA probes. In contrast with T. brucei brucei and T. brucei rhodesiense stocks, all the T. b. gambiense stocks are characterized by a conserved, specific DNA band pattern, regardless of the probe. This allows T. b. gambiense to be non-ambiguously identified. On the contrary, T.b. brucei and T. b. rhodesiense, which could not be discriminated by the same criteria, both yield highly variable DNA band patterns. Our data confirm that domestic animals like pig, dog and sheep constitute a potential reservoir for T.b. gambiense. Using a numerical analysis of the DNA hybridization patterns we have measured the degree of similarity between the 72 trypanosome stocks. This investigation shows that all T.b. gambiense stocks are included in the same homogeneous population, while the stocks from the two other subspecies seem to be distributed in several heterogeneous groups, some of these showing correlation with the geographical origin of the trypanosomes. It is concluded that (i) T.b. gambiense stands out as a real subspecies that has undergone a distinct evolution relative to the 'non-gambiense' group, (ii) the alleged T.b. rhodesiense subspecies does not fit with any of the groups evidenced by our cladistic analysis and hence does not appear as a distinct subspecies and (iii) 'non-gambiense' trypanosomes are probably evolving much more rapidly than T.b. gambiense. Different aspects of trypanosome relationships and evolution are discussed.