Some preliminary observations on the susceptibility and resistance of different cattle breeds to Theileria parva infection.

Theileria parva-naïve Friesian (Bos taurus), Boran (Bos indicus) and Maasai Zebu steers (B. indicus) were infected with a T. parva sporozoite stabilate dose which had previously been shown to induce an estimated 50% mortality rate in Boran cattle. All the cattle developed patent infections with no significant differences in the length of the prepatent period to development of macroschizonts (P > 0.05) between the three groups. Clinical theileriosis occurred in all eight the Friesians (100%), five out of nine Borans (55.6%) and two out of five Zebus (40%). Three of the Friesians (37.5%), and two of the Borans (22.2%) died of theileriosis. The different cattle types were equally susceptible to the infective dose used as indicated by the length of the prepatent periods, but there was a marked difference in their development of clinical theileriosis. The gradation in resistance to disease confirms the findings of earlier less critical studies and identifies these cattle breeds as suitable for investigations into the mechanisms of resistance to theileriosis.

Identification of a target population for immunisation against East Coast fever in coastal Kenya.

Two experiments were carried out to identify the target population of cattle for immunisation against East Coast fever (ECF) using the infection-and-treatment method. Firstly, a sentinel-calf study was used to determine the age window for ECF immunisation by determining ages at clinical detection of infection with Theileria parva. Six groups of five naive cross-bred (Bos taurus/Bos indicus) male calves, introduced at intervals of 2 months at a mean age of 26 days, were exposed to natural tick challenge on a high ECF-risk, small-holder farm in the coastal lowland, coconut-cassava agro-ecological zone of coastal Kenya. Secondly, a challenge study evaluated the relationship between the presence of T. parva antibodies and immunity. Ten indigenous adult Zebu cattle and nine Zebu young stock purchased from farmers in the same zone, and eight cross-bred calves (survivors of the sentinel-calf study) were challenged with 10 times the immunising dose of T. parva Marikebuni stock. Twenty-four of these 27 cattle had high antibody titres before challenge. Two cross-bred calves, obtained from an ECF-free area and seronegative to T. parva schizont antigen, also were challenged and used as susceptible controls. Twenty-five (83%) of the 30 sentinel calves contracted ECF over an age range of 36-116 days (mean 72 days). The remaining five calves died of other causes within 2 months of arrival on the farm. Fourteen of the 25 calves survived the infection and developed antibodies to T. parva. Despite tick control, seven of these 14 calves had a second episode of ECF and two died. In total, 13 of the 25 calves that contracted ECF died. Only one of 19 indigenous Zebu animals developed clinical ECF when challenged with T. parva Marikebuni (mild clinical signs with spontaneous recovery). Of the eight cross-bred survivors from the first experiment, only one succumbed to ECF when challenged and it died. Both susceptible cross-bred calves developed severe clinical signs of ECF and one died. The experimental studies show that in the high ECF-risk areas of the coconut-cassava zone of coastal Kenya, immunisation against ECF in cross-bred (B. taurus/B. indicus) cattle should be targeted at an early age (preferably within 1-2 months of birth).

Immunization of cattle against East Coast fever using Theileria parva (Marikebuni) and relaxation of tick control in North Rift, Kenya.

A total of 90 animals was immunized against East Coast fever (ECF) using Theileria parva (Marikebuni) stock on three large-scale farms in Kiminini Division, Trans-Nzoia District, North Rift, Kenya. Another 90 cattle served as non-immunized controls. Following immunization the number of cattle with significant indirect fluorescent antibody (IFA) titres increased from 43.9% to 84.4% and 6.7% of the cattle developed clinical ECF reactions. Two months after immunization, the immunized and non-immunized cattle were divided into two groups one of which was dipped every 3 weeks and the other dipped when total full body tick counts reached 100. All the animals were monitored for 51 weeks for incidences of ECF and other tick-borne diseases. Twenty-four cases of ECF were diagnosed among the non-immunized cattle compared to four cases among the immunized cattle; a difference that was significant (P > 0.05). There was no significant difference in the incidences of babesiosis and anaplasmosis between the immunized and non-immunized cattle.

Comparison of indirect fluorescent antibody test and enzyme linked immunosorbent assay in the detection of exposure of cattle to Theileria parva in Kenya.

Appraisal of the indirect fluorescent antibody test (IFAT) and antigen enzyme linked immunosorbent assay (ELISA) serological tests as carried out to detect cattle exposed to Theileria parva at the National Veterinary Research Centre, Muguga (NVRC), Kenya is reported. Using sera from T. parva naive cattle and cattle experimentally exposed to T. parva, the two tests were appraised in terms of their sensitivity and specificity. IFAT and ELISA had the same sensitivity of 90% while ELISA had a higher specificity (90%) than IFAT (80%). A comparison was also made of the capability of the two tests to detect exposure of dairy cattle to T. parva prior to immunization against East Coast fever (ECF). The positive outcome from the IFAT was significantly higher (chi 2 = 30.36; P < 0.001) than that from the ELISA. The agreement between the two tests was low (Kappa = 0.21). The two tests indicated a higher risk of ECF in the study area than was expected. Indications are that the ELISA has been effectively adopted at NVRC.

Cost analysis of immunisation against east coast fever on smallholder dairy farms in Kenya.

A spreadsheet model was developed and used to estimate the total cost of immunising cattle against East Coast fever (ECF) based on the infection-and-treatment method. Using data from an immunisation trial carried out on 102 calves and yearlings on 64 farms in the Githunguri division, Kiambu district, Kenya, a reference base scenario of a mean herd of five animals, a 10% rate of reaction to immunisation and a 2-day interval monitoring regimen (a total of 10 farm visits) was simulated. Under these conditions, the mean cost of immunisation per animal was US$16.48 (Ksh 955.78); this was equivalent to US$82.39 (Ksh 4778.90) per five-animal farm. A commonly reported reactor rate of 3% would decrease the cost of US$14.63 (Ksh 848.29) per animal. Reducing the number of farm monitoring visits from 10 to 7 would reduce the total cost by 10%, justified if farmers are trained to undertake some of the monitoring work. The fixed costs were 53% of the total cost of immunisation per farm. The cost of immunisation decreased with increasing number of animals per farm, showing economies of scale.

A mild Theileria parva parasite with potential for immunisation against East Coast fever.

Twenty-three Friesian cattle were inoculated subcutaneously anterior to the left prescapular lymph node with 1 ml of a mild isolate of Theileria parva. The cattle developed low macroschizont parasitosis but no clinical reaction was observed. Thirty-five days later the cattle were grouped into five groups and challenged with five different Theileria parva isolates (four cattle-derived Theileria and one buffalo-derived Theileria). The cattle were all solidly immune to challenge with the cattle-derived Theileria isolates but three out of five of the cattle challenged with the buffalo-derived parasite died of theileriosis. All ten non-immunised control cattle developed severe theileriosis and were treated with buparvaquone (Butalex; Pitman-Moore).

Immunisation of rabbits against the brown ear tick, Rhipicephalus appendiculatus using tick haemolymph.

Tick naive rabbits were immunised with haemolymph components from partially fed Rhipicephalus appendiculatus adult ticks and subsequently challenged with all the developmental instars of the tick. The results obtained showed that the rabbits were rendered resistant to all the instars of the tick. However, the resistance was more pronounced in adult ticks than in the immature stages. The resistance was manifested as a reduction in the number of ticks that fed successfully to engorgement, reduced engorgement weights and reduced fecundities. Re challenging the resistant experimental rabbits with all the developmental instar stages of Rhipicephalus appendiculatus showed that resistance was maintained in subsequent infestations.

Evaluation of infection and treatment methods in immunization of improved cattle against theileriosis in an endemic area of Kenya.

Five experiments were carried out to determine the efficacy of immunization against theileriosis in an endemic area of Kenya using artificial infection with a mixture of stabilates of Theileria parva stock or natural infection and treatment with parvaquone or several formulations of oxytetracyclines. For the first four experiments, introduced, susceptible Sahiwal/Friesian crosses were used and in the fifth, calves of Boran/Maasai zebu crosses born on the site. Cattle were infected either artificially with sporozoite stabilates of local isolates of T. parva parva derived from cattle and T. parva lawrencei derived from African buffalo or exposed to natural tick challenge on the ranch mostly derived from buffalo. The cattle were then given various treatment regimens using either parvaquone or long- and short-acting formulations of oxytetracycline. Treatment of natural infections, although it can be effective, was not considered a practical method on a large scale because of the need for intensive monitoring in the case of parvaquone treatment and the possibility of cattle not becoming infected in the case of prolonged application of long-acting formulations of oxytetracycline. Both methods were relatively expensive. Artificial infection treatment proved more practical and methods were developed where the monitoring of cattle was not required during the immunization procedure. Out of a total of 16 drug regimens investigated, one (consisting of two treatments of a short-acting formulation of oxytetracycline at 10 mg kg-1 body weight on Days 0 and 3 or 4 after infection) was found to be the most efficacious and the cheapest, and has now been used on a routine basis. This method can be used successfully on calves greater than 1 month of age.

Immunization of cattle using varying infective doses of Theileria parva lawrencei sporozoites derived from an African buffalo (Syncerus caffer) and treatment with buparvaquone.

A Theileria parva lawrencei isolate in the form of a sporozoite stabilate, derived by feeding clean Rhipicephalus appendiculatus nymphal ticks on an African buffalo (Syncerus caffer) captured in the Laikipia District, Kenya, was inoculated into groups of cattle at dilutions between 10(0) and 10(-3). Groups of 3 cattle infected with 1 ml inocula at 10(0), 10(-1) and 10(-2) dilutions were treated with 2.5 mg/kg body weight of buparvaquone on day 0 and similar groups were left untreated to act as controls. An additional group, given 10(0) dilution of the stabilate, was treated with buparvaquone on day 8 post-inoculation. It was found that all control cattle inoculated with the stabilate at dilutions between 10(0) and 10(-2) became infected, but only 2 out of 3 cattle developed patent infections at 10(-3) dilution. All 3 control cattle receiving 10(0) dilution died of theileriosis, 2 at 10(-1) and 10(-2) dilutions, and 1 at 10(-3) dilution died. Buparvaquone treatment on day 0 at 10(0) dilution resulted in the survival of 2 of 3 cattle and all the cattle at 10(-1) and 10(-2) dilutions. All the surviving cattle eventually developed a significant serological response against T. parva in the indirect fluorescent antibody test, except 1 in the 10(-3) dilution group, and were immune to homologous challenge when tested 3 months later with a lethal inoculum of stabilate, except 2 cattle in the 10(-3) dilution group. As a result of a theileriosis problem at about day 60 after inoculation in 2 cattle given 10(-2) dilution of stabilate and buparvaquone treatment on day 0, an additional 5 cattle were given 10(-2) dilution of stabilate and developed a good immunity after buparaquone treatment. None was shown to develop the carrier state. Treatment with buparvaquone on day 8 after infection with 10(0) dilution of stabilate was not successful since 2 died. The stabilate used was shown to produce reproducible infection in cattle at different dilutions.

Comparison of long acting oxytetracycline and parvaquone in immunisation against East Coast fever by infection and treatment.

Two groups of five cattle were immunised with a field isolate of Theileria parva as stabilate and simultaneously treated with long acting oxytetracycline or parvaquone in early clinical disease. The oxytetracycline group suffered a marked fall in leucocyte count and one animal died during immunisation. The parvaquone group suffered a less severe fall in leucocyte count and all survived. The surviving immunised cattle were immune to homologous challenge but susceptible to subsequent challenge with T p lawrencei.