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The current challenges of dourine: difficulties in differentiating Trypanosoma equiperdum within the subgenus Trypanozoon.

During its 20th annual meeting in Paris in May 1999, the OIE (World organisation for animal health) Ad Hoc Group on Non-Tsetse Transmitted Animal Trypanosomoses expressed the following concerns about dourine: the discrepancies in some of the results of the complement fixation test (CFT), which is the only international diagnostic test officially recognised by the International Organisation for the Transportation of Equidae; the persistence of suspected cases of dourine in some Asian, European and African countries; the impossibility of differentiating Trypanosoma equiperdum from Trypanosoma evansi and of isolating new strains of T. equiperdum from clinical cases that have appeared in various parts of the world since 1982. In the light of these concerns, it was decided, in agreement with the Directorate of the Federal Veterinary Services of Russia in Moscow, to perform comparative trials on the value of CFT/dourine at the OIE Reference Laboratory for dourine in Moscow (The All-Russian Research Institute of Experimental Veterinary Medicine) using reagents (antigens and sera) from seven countries with extensive experience in the field of dourine diagnosis, namely, South Africa, France, Italy, Germany, Russia, the United States of America and the People's Republic of China. It is thanks to the successful co-operation of these countries that the trials were made possible. Results showed an overall concordance and were submitted for consideration to the OIE Biological Standards Commission, the commission which is in charge of the Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. These trials serve as a starting point for further study, particularly in the following areas: the isolation of new strains of T. equiperdum from clinical dourine cases; the identification of specific markers for T. equiperdum which would make it possible to differentiate it from among the other species within the subgenus Trypanozoon; the experimental infection of horses with newly isolated T. equiperdum strains to compare their pathogenicity with those currently used in national diagnostic laboratories and with that of T. evansi; phylogenetic studies; the proposal and validation of new, internationally recognised diagnostic test(s) for dourine.

Animals↗

Detection of trypanosome DNA in serologically positive but aparasitaemic horses suspected of dourine in Ethiopia.

A field study of horses was conducted in the province of Bale, Ethiopian highlands. A rapid questionnaire analysis indicated that dourine, known as "Dirressa", is a major health problem of equines in this area. A total of 121 horses suspected of dourine were examined by use of clinical, parasitological, serological and DNA based techniques. Incoordination of hindlegs (76%), swelling of external genitalia (48.8%) and emaciation (39.7%) were the most common clinical signs observed. Using the haematocrit centrifugation technique (HCT), no trypanosomes were detected in blood, genital washes or tissue fluids. By contrast, trypanosome specific DNA products were amplified by PCR and subsequently detected by DNA probe hybridization in blood samples of 29 horses (29/104), all serologically positive by CFT and/or ELISA. Positive PCR results were significantly associated with swelling of external genitalia (P< 0.05). There is strong evidence, although there was no direct detection of T. equiperdum, that dourine is highly prevalent in the area, a finding which is in accordance with earlier reports. It is concluded, that this PCR assay provides a very sensitive tool in the diagnosis of active infections of dourine in endemic areas where trypanocidal drug use is common.

Animals↗

Apparent prevalence of dourine in the Khomas region of Namibia.

A 15-year record of the results of horse sera from the Khomas region of Namibia tested by the complement fixation test for dourine at the Central Veterinary Laboratory in Windhoek before clearing the respective animals for export and competitive sport were subjected to statistical analysis. The range of percentage positive, taken as the apparent prevalence of dourine for the region, during the period of study, was 0-29.09%; the average regional level of apparent prevalence was 8.33%. These figures were thought to be lower than the real situation due to some bias in the sampling criteria. For more accurate results, the more reliable enzyme-linked immunosorbent assay techniques are recommended for use in sero-surveys for dourine in Khomas and other regions of Namibia to provide a basis for development of effective control strategies against the disease.

Animals↗

The use of a single complement fixation test technique in bovine brucellosis, Johne's disease, dourine, equine piroplasmosis and Q fever serology.

The same techniques may be used in the complement fixation test (CFT) for the serological diagnosis of bovine brucellosis, Johne's disease (paratuberculosis), dourine, equine piroplasmosis and Q fever (caused by Coxiella burnetii). The reproducibility of results is excellent, falling for the most part within the twofold range and never exceeding the fourfold range. Agreement with other laboratories is excellent (i.e. within twofold) in the case of brucellosis and equine piroplasmosis antibody titres. A good correlation between the occurrence of the disease and serological reactions is found on circumstantial evidence in the cases of dourine, Johne's disease and Q fever. A standard unitage system is used to report the antibody titres found in all the tests. To simplify laboratory protocols, laboratories required to employ the CFT for the diagnosis of these diseases are advised to use a single proven technique in all the tests. Problems experienced with transient false-positive Johne's disease antibody titres in cattle following on tuberculin (bovine and avian) testing make it advisable to take specimens for the Johne's disease test prior to performing the tuberculin tests.

Animals↗

Evaluation of agar gel immunodiffusion and indirect fluorescent antibody assays as supplemental tests for dourine in equids.

The agar gel immunodiffusion (AGID) and indirect fluorescent antibody (IFA) assays were evaluated as supplemental tests to the complement-fixation (CF) test, the official US importation certification test for dourine in equids. The American stabilate (n = 10 animals) or the Canadian stabilate (n = 6 animals) of Trypanosoma equiperdum cultured in rat blood was administered by catheterization and infusion in the urogenital tract of 16 equids. To assess parasitemia and serologic responses by use of the CF, AGID, and IFA tests, a total of 787 serum and blood samples were obtained from equids before exposure and 3 times a week after exposure to T equiperdum. Results of the IFA and AGID tests were compared with the CF test results. The disease was diagnosed earlier by the IFA test than by the AGID test, regardless of antigen preparation or exposure group. The mean number of days between exposure and positive result by the CF and IFA tests was the same when either homologous or heterologous antigen was used in the IFA test. In general, the IFA test was more sensitive than the AGID test in diagnosing dourine, regardless of the antigen preparation used in the test or exposure group. Differences in test specificity were observed among both groups of exposed equids when either antigen was used (P < 0.05). The AGID test, using the American antigen, was more specific than the IFA test for sera from both groups of equids. When the Canadian antigen was used, the IFA test was a more specific test than the AGID test (P < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Agar↗

Observations on the transmission, immunology, clinical signs and chemotherapy of dourine (Trypanosoma equiperdum infection) in horses, with special reference to cerebro-spinal fluid.

This paper is a record of observations on the transmission and clinical signs of dourine in naturally infected cases of known duration, and of temporal and quantitative aspects of the immune response in blood and cerebro-spinal fluid. Included in the record are observations on the presence of Trypanosoma equiperdum parasites in these body fluids and methods for their detection. There is evidence that the occurrence of nervous symptoms and lesions in infected horses is associated with the presence of Trypanosoma equiperdum parasites in cerebro-spinal fluid. The suitability of cerebro-spinal fluid as an environment for the parasite and its relationship with nervous manifestations of the disease are discussed. Observations support the previously reported lesions of peripheral polyneuritis and suggest a possible correltation between the consitstent position of the nervous lesions and the drainage of cerebro-spinal fluid containing the parasite. Chemotherapy with an experimental drug MSbE was used with varying results in 4 horses at different stages of infection.

Anemia↗

An investigation into alternative methods for the serodiagnosis of dourine.

The complement fixation test (CFT), indirect fluorescent antibody test (IFAT), card agglutination test for trypanosomiasis (CATT) and enzyme-linked immunosorbent assay (ELISA) were compared in their application to the serological diagnosis of Trypanosoma equiperdum infection in 43 horses. The CFT remains a reliable test for dourine, especially in countries where other members of the subgenus Trypanozoon do not occur. The IFAT is a good 'back-up' test, but, requiring skilled operators it has the disadvantage of making it labour intensive, and interpretation of results subjective. This makes it more suited to small numbers of samples. The ELISA is suitable for large numbers of samples and could readily be used in routine diagnostic procedures. The CATT could be of value in field situations, although it does not appear to be as sensitive as the CFT. Its possible application under these conditions should be further investigated.

Agglutination Tests↗

Dourine in Southern Africa 1981-1984: serological findings from the Veterinary Research Institute, Onderstepoort.

The distribution of positive dourine cases found on the complement fixation test at the Veterinary Research Institute, Onderstepoort from 1981 to 1984, is recorded. Within the Republic of South Africa, foci of infection occurred in the Johannesburg, Pretoria, Potchefstroom, Rustenburg, Upington, Lichtenburg, Kroonstad, Louis Trichardt, Middelburg (Cape) and Mossel Bay state veterinary districts. In Bophuthatswana, Transkei, Lesotho, South West Africa and Swaziland, positive cases were also recorded. Anti-complementary activity of horse sera does not present a problem. In donkey and mule sera, however, sera show anticomplementary activity in approximately 50% of cases. This can be reduced by diluting the sera 1/2 before inactivating at 63 degrees C for 1 h.

Animals↗

The prevalence of dourine in southern Africa.

The geographical distribution of positive dourine cases found on serological testing at Onderstepoort during the period 1954 to 1975 is recorded, showing major foci of infection occurring in the northern Cape Province, northern Orange Free State, western and north-western Transvaal. Positive cases are also recorded from South West Africa, Botswana, Lesotho, Swaziland and Rhodesia.

Africa, Southern↗

Comparative evaluation of enzyme-linked immunosorbent assay (ELISA) for the serodiagnosis of dourine.

The detection of antibodies against Trypanosoma equiperdum in 689 equid sera was compared by enzyme-linked immunosorbent assay (ELISA), the complement fixation test (CFT) and an indirect immunofluorescent test (IIF). CFT was the least sensitive technique, susceptible to anti-complementary factors and the most technically demanding. IIF was more sensitive, but was only suitable for testing limited numbers of samples. In this study, ELISA was the most sensitive test, the least labour intensive and lends itself to a considerable degree of automation. It is suggested that ELISA would be relatively easy to standardise between laboratories and an ELISA protocol could be adopted as the internationally approved test for equine health certification.

Animals↗