Development and evaluation of an enzyme-linked immunosorbent assay (ELISA) for the determination of the trypanocidal drug homidium in serum of treated cattle

Two enzyme-linked immunosorbent assays (ELISA) for the determination of homidium in serum of treated cattle have been developed and evaluated. One is a direct competition (Assay 1) and the other an indirect competition assay (Assay 2). Both assays are highly sensitive with a limit of detection of 0.1 ng homidium per mL serum. Homidium levels were measurable in serum of cattle for over 2 months following administration of a single intramuscular (i.m.) dose at 1 mg/kg bodyweight. The level of sensitivity afforded by these assays makes them potentially useful tools in the pharmacokinetic evaluation of homidium and for investigating drug resistance or causes of drug failure. Assay 2 was chosen as being most suitable for further studies.     

Application of the Vettest 8008 system for the biochemical analysis of vervet monkey plasma

A clinical biochemistry analyser designed specifically for veterinary use was used to analyse plasma samples from 24 vervet monkeys (Cercopithecus aethiops). Two millilitres of heparinised blood was collected from each of the 24 monkeys on four occasions at intervals of one week. Plasma was separated and analysed for the concentrations of triglycerides, cholesterol, total proteins, albumin, globulins, creatinine and blood urea nitrogen (BUN) and the activities of alkaline phosphatase (AP), lactate dehydrogenase (LDH), alanine aminotransferase (ALT), aspartate aminotransferase (AST), and creatine kinase (CK). The tests were easy to perform, used small volumes of plasma, and yielded consistent results for most of the analytes. The activities of CK and AP, but not AST, appeared to be influenced by haemolysis, and there were significant individual variations in the activity of LDH.     

New mosquito larvicidal tetranortriterpenoids from Turraea wakefieldii and Turraea floribunda

The crude methanol extracts of the root barks of Turraea wakefieldii and Turraea floribunda were found to show mosquito larvicidal activity against third-instar larvae of Anopheles gambiae sensu stricto. Four new limonoids comprising a vilasininoid 1 and three havanensinoids 2-4 were isolated from the chloroform fractions of the methanol extracts of T. wakefieldii and T. floribunda, respectively. The structures of the compounds were elucidated by NMR spectroscopy. Compounds 1, 2, and 4 had LD50 values of 7.1, 4.0, and 3.6 ppm, respectively, and were more potent than azadirachtin, which had an LD50 value of 57.1 ppm when tested against larvae of A. gambiae.     

Preliminary field test of lyophilised contagious caprine pleuropneumonia vaccine

Fifty goats were immunised in the field against contagious caprine pleuropneumonia (CCPP) using a single dose (0.15 mg) of lyophilised, saponin killed Mycoplasma strain F38. Two months after vaccination, these goats together with 50 unimmunised control goats were challenged by contact with goats experimentally infected with CCPP. Twelve vaccinates and 14 controls died of diarrhoea due to salmonella infection during the first two weeks after challenge. The remaining immunised goats (38) with the exception of two goats which had elevated temperatures were protected from CCPP. Of the remaining 36 control goats, 30 contracted CCPP at a mean of 39 (+/- 14.3 SD) days after challenge and 27 of them died. These results show that the lyophilised killed F38 vaccine conferred 100 per cent protection against mortality and 95 per cent protection against clinical disease caused by Mycoplasma species strain F38.     

Growth and Mortality in Sheep and Goats under High Tsetse Challenge in Kenya

Trypanosomosis is a major impediment to livestock production and economic development in those areas of Africa where it is endemic. Although small ruminants appear to perform better than cattle in various agro-ecological zones, the importance of trypanosomosis has not been extensively investigated in these livestock. This study was designed to investigate the prevalence of trypanosomosis in sheep and goats in an endemic area and to evaluate the performance of different breeds under high tsetse challenge and the potential role of chemoprophylaxis in the control of the disease. The results showed that tsetse flies feed readily on small ruminants, and that these animals are susceptible to trypanosomosis. The Small East African goats acquired fewer infections than the Black Head Persian and Dorper sheep used in the study. In both sheep and goats, chemoprophylaxis with isometamidium chloride (Samorin, Rhone Merieux, Annecy, France) was protective, resulting in fewer infections and higher body weight gain. Trypanosomosis caused anaemia in both sheep and goats, and animals whose PCV fell below 15% rarely recovered, even with trypanocidal drug treatment. The peak transmission period was between 1 and 3 months after the peak tsetse fly density, which raises the possibility of effective strategic prophylaxis.     

Financial analysis of animal trypanosomiasis control using cypermethrin pour-on in Kenya

The financial impact of use of cypermethrin pour-on (Ectopor^R) in control of animal trypanosomiosis was determined in a trial undertaken by the Kenya Trypanosomiasis Research Institute (KETRI). This trial started in December 1990 and ended in February 1992. It was undertaken in two adjacent ranches in the coast province of Kenya. The trial site was in an area of high apparent density (AD) of tsetse flies, and at the start of the trial no cattle were kept in this area. Cypermethrin was applied fortnightly to the 1100 steers which were kept in pour-on ranch ‘A’ while another 100 steers were kept in control ranch ‘B’ to act as control sentinels. From the main pour-on group, 100 animals were identified as the pour-on sentinels and compared to the control sentinels which received no pour-on.

Pour-on application led to a significant decrease in the tsetse AD in the pour-on ranch A to 90% of the initial AD in some areas. The animals treated with pour-on had a significantly higher mean packed-cell volume (PCV). The weekly prevalence of trypanosome infections in animals treated with pour-on was <4% with only one exception when it was <10%. In the control animals, the prevalence ranged between 10 and 50% (with a few exceptions when it was <10%). The incidence of tick-borne diseases was lower in the pour-on animals. The mean monthly weights of the pour-on animals was significantly higher, and at the end of the trial the pour-on animals had a mean weight gain of 136.70±16.7 kg while the control animals had gained 97.16±22.6 kg.

The financial net return of using cypermethrin pour-on was positive and the financial rate of return of 122.6% indicated that use of the pour-on was highly beneficial despite the high cost of the product.     

Cardiac damage in dogs infected with T brucei; clinical and electrocardiographic features

Intravenous infection of dogs with Trypanosoma brucei resulted in an acute disease syndrome, characterised by fever, intense parasitaemia, severe anaemia and rapid weight loss. During the course of infection, evidence of severe cardiac abnormalities developed. Tachycardia and tachypnoea occurred soon after detection of parasitaemia, around day 6, progressing to severe bradycardia and dyspnoea in terminal stages of the disease in week 4. Murmurs of mitral and tricuspid incompetence were heard from day 12 by auscultation of the thoracic cavity. Electrocardiography revealed abnormalities in generation and conduction of electrical impulses, including sinus arrest and atrioventricular blocks, and accumulation of pericardial effusion in terminal stages. Effective treatment with the trypanocidal drug suramin resulted in rapid improvement of one of the dogs. When treatment was unsuccessful, however, chronic myocardial damage developed, with intramyocardial conduction defects, including S-T segment and T wave changes, and ventricular escape beats. These abnormalities were similar to those reported for human African trypanosomiasis.     

Characterisation of Theileria parva isolates from Kiambu district, Kenya

Four Theileria parva isolates from Muguga area of Kiambu district, Kenya, were used to establish schizont-infected cell lines. Their protein antigens were then separated by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS page). The isolates were subsequently subjected to protein analysis and characterisation by the western immunoblotting technique. Probing for the polymorphic immunodominant molecule (PIM) was done using monoclonal antibody no. 4. SDS page detected up to 20 protein antigens of molecular mass 35,000-180,000 Da. The western blot analysis revealed a greater heterogeneity in the molecular mass (M(r)) of PIM than previously thought. The M(r) of PIM varied between 80 and 90 kDa. The isolates further revealed different densities of surface epitopes with variable reaction to the monoclonal antibody. The implications of these findings to the epidemiology of east coast fever and immunisation programmes are discussed.     

Standardised tests in mice and cattle for the detection of drug resistance in tsetse-transmitted trypanosomes of African domestic cattle

Resistance to the drugs used to control African animal trypanosomosis is increasingly recognised as a constraint to livestock production in sub-Saharan Africa. The most commonly used tests for detection of trypanocidal drug resistance are tests using mice or ruminants, but these suffer from lack of standardisation and hence it may be difficult to compare the results of different investigators. Tests in mice are less expensive than tests in ruminants, but while tests in mice they may be useful as a general guide to resistance in a geographic area they should not be extrapolated to cattle on an individual trypanosome level. Moreover, the commonly used protocols are too laborious for their application to large number of trypanosome isolates on an area-wide basis. This paper presents guidelines for standardised testing of trypanocidal drugs in vivo, and introduces a simplified single-dose test for use in mice, which is convenient for use in areas with limited laboratory facilities. The single-dose test is appropriate for characterisation of geographic areas in terms of trypanocidal drug resistance using large numbers of trypanosome isolates, for making comparisons between areas, and for monitoring changes in trypanocidal drug resistance over time. Multiple-dose tests may be used to determine the degree of resistance of individual stabilates to be determined precisely in mice are also described, but for logistical reasons these will rarely be conducted on more than a few stabilates, and testing of a larger number of stabilates in the single-dose test will generally provide more useful information. Finally, we describe tests in cattle that may be used to determine the efficacy of recommended curative doses of trypanocidal drugs for the treatment of infection with individual trypanosome isolates, including Trypanosoma vivax, which is rarely infective for mice.     

Rinderpest Vaccination and the Incidence and Development of Trypanosomosis in Cattle

An investigation was made into whether recent vaccination of cattle with tissue culture rinderpest virus would cause immunosuppression and lead to more frequent or more severe infection with trypanosomes in animals grazing in tsetse-infested areas. Herds of cattle on Galana Ranch in Kenya were divided, with approximately half of each herd being vaccinated with tissue culture rinderpest virus strain Kabete O, while the rest remained unvaccinated. The herds were then exposed to the risk of natural infection with trypanosomes on the ranch. Three experiments were performed during different seasons. Infections with Trypanosoma congolense and Trypanosoma vivax were frequently detected but there was no evidence that vaccinated animals were more likely to acquire trypanosome infections or to show a more severe disease than unvaccinated cattle. It is concluded that tissue culture rinderpest vaccine does not cause immunosuppression and can safely be used in cattle likely to be exposed to tsetse flies and trypanosomosis.