Age resistance to Theileria parva bovis infection in calves.

Data collected in the Zimbabwean province of Mashonaland-West, in the period 1980-1988, showed that mortality in calves owing to Theileria parva bovis infection (January disease) was significantly lower in animals younger than 7 months than in older cattle. Groups of seven Holstein-Friesian calves from non-immune dams aged approximately 1, 4, 7, 10 and 13 months were infected with a Theileria parva bovis tick-derived stabilate. The dose chosen was lethal for 40% of the calves in the trial. Mortality was highest in the 4-month age group. The reactions in the 7-, 10- and 13-month age groups became progressively milder. The reactions in the 1-month old calves were the least marked, being very mild. The age-related resistance in the youngest calves, as can be concluded from our results, is only of short duration and cannot explain the lower incidence of January disease observed in calves in the field.

Geographical information systems for studying the epidemiology of cattle diseases caused by Theileria parva.

Data on selected variables which influence the epidemiology of cattle diseases caused by Theileria parva were assembled and entered in a computerised geographical information system. Variables studied included the distributions of major hosts (cattle and buffalo), the vector ticks (Rhipicephalus appendiculatus and related species) and the reported presence of East Coast fever, corridor disease and January disease. In addition, the distribution of climatic suitability for R appendiculatus was assessed using the model CLIMEX run on an interpolated climate database developed for Africa. Distribution maps of each variable were produced. The potential value of geographical information systems in studies of disease epidemiology and control is discussed, with examples of how sensitivity may be enhanced by the inclusion of additional variables. In addition, subject areas in which poor data quality and inadequate data standardisation may limit the use of these systems are identified and discussed.

Financial analysis of East Coast fever control strategies on beef production under farm conditions.

This study provides a financial analysis of an East Coast fever immunisation trial conducted on a farm in the Coast Province of Kenya. The objective of the trial was to assess the effects of immunisation by the infection and treatment method, under different acaricidal treatments, on the productivity of beef cattle. Eighty beef cattle were immunised and an equal number acted as controls. The immunised and unimmunised groups were divided into four subgroups of 20 animals. Two subgroups, one from the immunised and one from the unimmunised group, were sprayed with acaricide twice a week; a second pair of subgroups was sprayed once every three weeks; a third pair had prolonged release acaricide-impregnated ear-tags inserted into each ear; and a fourth pair had no tick control treatment. Financial analysis revealed that the immunised subgroups were more profitable, owing to lower mortality and higher weight gains than the unimmunised subgroups. Of the immunised subgroups, the best was that sprayed with acaricide twice a week; it yielded a marginal rate of return of 244 per cent and maximised financial benefits to the farmer in this trial. However, further trials under different production circumstances would be required before the method could be recommended for widespread adoption.

Immunization of cattle with a Theileria parva bovis stock from Zimbabwe protects against challenge with virulent T.p. parva and T.p. lawrencei stocks from Kenya.

Following inoculation of 34 Bos indicus (Boran) cattle with a Theileria parva bovis (Boleni) stock from Zimbabwe, 18 animals underwent mild theilerial reactions, 12 underwent moderate reactions, three suffered severe reactions and one died. When these animals were subsequently challenged with different virulent stocks of either T.p. parva (Muguga, Marikebuni or Mariakani) or T.p. lawrencei (Ngong 1 or Nanyuki) from Kenya, all except two animals resisted challenge. The two reactors were part of the group challenged with the T.p. parva (Mariakani) stock. All 12 susceptible control animals underwent severe reactions and 11 died. The results of these experiments suggest that T.p. bovis (Boleni) may be used in some situations to immunize cattle against East Coast fever without the need to provide concomitant chemotherapy as in the infection and treatment method of immunization.

Long-acting oxytetracycline prophylaxis to protect susceptible cattle introduced into an area of Kenya with endemic East Coast fever.

Two field trials were carried out in successive years at the Ngong Veterinary Farm, Kenya, in which young cattle, previously unexposed to tick-borne diseases, were introduced into an area with endemic East Coast fever while protected by a series of injections of a long-acting oxytetracycline. In 1984, 12 animals which received injections of 20 mg/kg of the drug on days 0, 7, 14 and 21 after introduction, together with 12 untreated controls, were exposed without tick control until clinical disease occurred. All 12 control animals contracted East Coast fever by day 24 and 10 of them died. Five of the 12 injected animals had detectable parasites, and one of them required antitheilerial treatment. In 1985, four groups of 10 calves were introduced. One group received injections of 20 mg/kg of oxytetracycline on days 7 and 14, one group received injections on days 7, 14 and 21, and a third group received injections on days 7, 12 and 17; the fourth group (controls) had no treatment until clinical disease occurred. By day 35 all the control animals had contracted the disease and one had died despite antitheilerial treatment. Three injections of oxytetracycline suppressed the disease so that mild reactions occurred in only four animals in each group, but two injections failed to prevent severe reactions in two animals and mild reactions in four others.(ABSTRACT TRUNCATED AT 250 WORDS)

The effect of East Coast fever immunisation and different acaricidal treatments on the productivity of beef cattle.

A trial was performed on a farm in the Coast Province of Kenya to study the effects of East Coast fever immunisation and different acaricidal treatments on the productivity of immunised and unimmunised beef cattle. Eighty cattle were immunised against Theileria parva parva (Marikebuni) by the infection and treatment method and a similar group was left as an unimmunised control. Immunisation had no deleterious effect on the cattle. After immunisation, the immunised and control groups were each subdivided into four groups of 20 and each subgroup was managed under a different tick control regimen. The tick control regimen were, acaricidal spraying twice a week or once every three weeks, the application of acaricide-impregnated ear-tags, and no tick control. During a nine-month exposure period there were 18 cases of East Coast fever among the 80 immunised cattle, three which were severe and the others mild. Among the 80 unimmunised cattle there were 57 cases of East Coast fever, 50 of which were severe. The highest morbidity and mortality occurred in the groups under limited tick control or without tick control. Overall weight gain in the immunised cattle, irrespective of the tick control regimen, was better than the weight gain in the unimmunised groups. Within the immunised groups, the weight gain of the cattle sprayed twice weekly was comparable to the weight gain of the animals with acaricidal ear-tags and was significantly higher than the weight gains in the groups sprayed once every three weeks or with tick control. Preliminary cost/benefit analysis showed that it was uneconomical to maintain unimmunised cattle under limited or no tick control.(ABSTRACT TRUNCATED AT 250 WORDS)

Isolation and characterization of bovine Theileria parasites in Zimbabwe.

Theilerial parasites of cattle were isolated by a variety of methods from the Harare area of Zimbabwe. Parasite stocks were established in lymphoid cell cultures and as cryopreserved sporozoite stabilates in the laboratory. Fourteen stocks in culture were characterized by testing them with monoclonal antibodies (MAb) raised against T. parva parva and T. parva lawrencei antigen. Two of these stocks had profiles similar to T. taurotragi isolates from East Africa, the other stocks had profiles similar to T. parva parva, however, many of them failed to bind MAb No. 7, and this may be a distinctive feature for T. parva bovis. Three T. p. bovis stocks were titrated by injecting different doses of the respective stabilates into pairs of cattle. Reactions ranged from severe to inapparent according to the stocks and dose used, but no fatal reactions were recorded, even at the highest dose rate. On recovery, all cattle were given homologous and then heterologous challenge. The results of the latter challenge showed that the Boleni stock gave good cross-protection against challenge with two other Zimbabwean stocks. This stock may therefore be a candidate for immunizing cattle, under field conditions, to protect them against T. p. bovis in Zimbabwe. Non-pathogenic strains of T. p. bovis may be difficult to distinguish from T. taurotragi unless cross-challenge experiments can be conducted and/or MAb profiles have been made. An improved serological test is needed to differentiate antibodies to these parasites in the sera of recovered cattle.

Effect of timing and intensity of challenge following immunization against East Coast fever.

When groups of Theileria parva parva Muguga-immunized cattle were given a homologous lethal challenge at different times after immunization, it was found that 4/6, 5/6, 6/6 and 6/6 animals survived when challenged on Days 5, 10, 20 and 30, respectively, post-immunization. With a heterologous challenge (T.p.parva Marikebuni), 2/6, 5/6, 4/6, 4/6 and 5/6 cattle survived when challenged on Days 5, 10, 20 and 30, respectively, after immunization. All controls, except one, died of East Coast fever (ECF). The survivor underwent severe ECF and recovered after a prolonged convalescence. When two T.p.parva Muguga-immunized animals were each given homologous challenge by application of 1000 infected ticks (infection rate of 20 infected acini (i.a.) per tick), both survived a mild ECF reaction. When groups of T.p.parva Muguga- or T.p.parva Muguga/Marikebuni-immunized cattle were challenged with different doses of T.p.parva Muguga sporozoites (equivalent of 140, 1400 and 14,000 i.a. per animal), 28/29 cattle survived. All controls died of ECF. It was concluded that cattle could be safely exposed to tick challenge 1 week after immunization by infection and treatment using appropriate immunizing stock(s). Massive homologous challenge did not break through the immunity induced by the immunization procedure.

Isolation of Theileria parasites from African buffalo (Syncerus caffer) and characterization with anti-schizont monoclonal antibodies.

Antigenic differences between intra-lymphocytic theilerial parasites isolated from the blood of 18 African buffalo and grown in vitro were assessed with anti-schizont monoclonal antibodies (mAbs). There was marked antigenic diversity both between isolates from different buffalo and between isolates taken at different times from the same buffalo. Many of the isolates from both wild and captive buffalo appeared to consist of mixed parasite populations. Some isolates were found by limiting dilution cloning and mAb testing to contain at least 3 or 4 distinct populations of Theileria. Once cloned, Theileria-infected lymphoblastoid cell lines retained their mAb profiles during prolonged in vitro cultivation and, when recloned, the subclones had the same mAb profile as their parent clone. The implications of these results for further studies on buffalo-derived theilerial parasites are discussed.

Development and survival of Theileria parva parva in Rhipicephalus appendiculatus exposed in the Trans-Mara, Kenya.

Nymphal Rhipicephalus appendiculatus (Trans-Mara) were fed on a steer infected with a Theileria parva parva (Kilae 1) stock isolated from an indigenous steer in the Trans-Mara Division, Kenya, which had a high piroplasm parasitaemia. A total of 5000 engorged nymphs which had dropped on one day were enclosed in elongated nylon bolting silk tubes in groups of 200-300 and were transported immediately to the Trans-Mara where they were suspended vertically in the grass cover with one end touching the ground. Over 98% of the nymphs moulted into adult ticks and 50% moult occurred by day 28 after exposure. The ticks showed over 80% survival up to 308 days post-exposure but thereafter showed a marked mortality so that only 22.5% of the ticks were alive after 439 days. Theileria parasites were detected in the salivary glands by day 35 post-exposure and infection rates and levels increased markedly between 180 and 235 days post-exposure. Thereafter, the infection rates and levels generally decreased. Groups of ticks were triturated and the resultant supernatant fluid inoculated into pairs of susceptible cattle, and these proved infective from day 44 to 145 after exposure. Three subsequent attempts to induce infections with supernatant fluid were unsuccessful. From 294 days after exposure, groups of 50 ticks were applied to cattle and caused lethal T. p. parva infections up to 439 days post-exposure. Climatic observations showed a relatively even monthly rainfall as well as mean maximum and minimum monthly temperatures.(ABSTRACT TRUNCATED AT 250 WORDS)

Clinical trial of halofuginone lactate for the treatment of East Coast fever in Kenya.

A field trial was undertaken to test the efficacy of halofuginone lactate in the treatment of East Coast fever under farming conditions in Kenya. The drug was administered orally at a dose of 1.2 mg/kg bodyweight and treatment was repeated after 48 hours. Of 293 cases treated 236 (80.5 per cent) recovered and 49 (16.7 per cent) died. Five animals were disposed of by the owners and three became chronically infected. No differences were recorded in recovery rate between uncomplicated cases and cases with concurrent anaplasmosis or babesiosis, nor were there any significant differences in recovery rates between animals of different types. However, zebu cattle (Bos indicus) tended to respond more rapidly to treatment than exotic cattle (Bos taurus). Younger animals had a poorer recovery rate than adults. Early treatments were more successful than those administered late. It was concluded that with early detection and treatment, coupled with efficient tick control, halofuginone lactate is effective in the treatment of clinical East Coast fever under field conditions.

Demonstration of colonies of Cowdria ruminantium in midgut epithelial cells of Amblyomma variegatum.

The development of colonies of Cowdria ruminantium was studied in midgut epithelial cells of adult Amblyomma variegatum that had become infected by feeding as nymphs on cattle with experimentally induced heart-water disease. Colonies were not observed in gut tissues obtained from nymphs during the feeding period, but were present in midgut epithelial cells of ticks obtained at 15 days after they were replete through molting to the adult stage. Colonies were small (1 to 10 micron) initially, but as tick development progressed, their diameter increased to as much as 60 micron. With electron microscopy, colonies were observed to be membrane bound and contained pleomorphic organisms that were reticulated. The organisms seemed to be dividing by binary fission. Many colonies contained a large, electron-dense inclusion that was morphologically similar to hemoglobin deposits found in the cytoplasm of midgut epithelial cells of recently fed ticks. Cowdria ruminantium was often observed adhered to these inclusions.

A life-cycle stage-specific antigen of Theileria parva recognized by anti-macroschizont monoclonal antibodies.

Four monoclonal antibodies, raised against macroschizonts of Theileria parva, were studied to characterize their antigen binding specificity. The indirect fluorescent antibody test showed that the antigen(s) recognized were present in the macroschizont stage of the parasite life-cycle but not in piroplasm, kinete or sporozoite stages. Immunoblot analysis of macroschizont stage antigens suggested that all four antibodies recognized the same antigen. This was a molecule which varied in molecular mass between different parasite stocks, ranging from 68,000 to 95,000 Da. The antigen was localized by immunoelectron microscopy to the surface of the intracellular macroschizonts.

Immunization against East Coast fever: the use of selected stocks of Theileria parva for immunization of cattle exposed to field challenge.

Two antigenically different stocks of Theileria parva parva (Kilifi and Marikebuni), previously characterized as belonging to groups A and C respectively on monoclonal antibody (MAb) profiles, were selected for immunization of different breeds of cattle against East Coast fever (ECF) by the infection and treatment method. A total of 52 immunized cattle and 33 susceptible controls of different group sizes were exposed to field challenge by ticks for periods of 42-90 days at three field sites where ECF is endemic on the Kenyan coast. All immunized cattle survived ECF challenge, but 87% of the controls died of the disease. The cattle exposed at one site had been immunized 1 year earlier and maintained tick-free in the intervening period. The level of immunity in these cattle was similar to that of cattle which had been immunized 1 or 2 months prior to exposure. Thus, immunity had not waned over the 1-year period. A study at another site showed that acaricidal treatment of immunized cattle could be safely extended from twice a week to once every three weeks, whereas in susceptible cattle even twice weekly spraying did not control ECF. The isolates made from infected controls during the trials indicated the presence of three T. p. parva stocks as defined by MAb profiles. Of the two stocks used for immunization, T. p. parva Marikebuni induced broader protection. In view of the apparent limited antigenic diversity of T. p. parva strains within the Coast Province it is suggested that the Marikebuni stock might represent a key stock for vaccination in this area.

Characterization of species and strains of Theileria.

A variety of methods is now available for characterizing species and strains of Theileria. For many practical purposes involving field control of theileriosis, characterization on a broad basis may be sufficient, but in other areas much more precise characterization is required. Such precision can be usefully exploited only when cloned parasite populations are involved, and methods to improve parasite characterization and parasite cloning should be developed concurrently. The current methods of immunization against theileriosis involve the use of live parasite populations which are generally poorly defined and, in addition, have the capacity to undergo biological change (by selection, mutation or genetic recombination) within hosts and vectors. Such changes may be difficult to define and identify, but could have profound effects on immunization strategies. Improved methods of parasite characterization and selection, which are now becoming available, will enable parasite stocks for immunization to be identified and selected more precisely, and any biological changes that occur can be monitored. Improved methods of parasite characterization will also open the way to a better understanding of Theileria genetics and the mechanisms of heritability, which appear to differ in some fundamental ways from patterns of Mendelian inheritance. Controlled matings between selected and defined populations of parasites can be envisaged, with the aim of producing hybrid parasites for immunization. In addition, the prospects of modifying the theilerial genome by genetic manipulation become very real: transfection vectors tailored by restriction enzymes could be used to insert or modify gene sequences to develop parasites with appropriate sets of characters. It may also be possible to identify parasite genes which trigger the cytotoxic response which is so important in immunity (Eugui and Emery, 1981; Emery et al., 1981; Preston et al., 1983). Such genes might then be transfected into bovine host lymphocytes to generate immunity against the whole parasite (Iams, 1985). The gene products which are responsible for stimulating immune responses could also be synthesized artificially and used for vaccination. Methods of characterizing Theileria range from Giemsa's staining to DNA hybridization; all have a role to play, and by judicious selection of appropriate methods for particular circumstances, it is becoming possible to characterize theilerial parasites very precisely. Improved methods of characterization can, in turn, lead to a better understanding of parasite biology and to the development of improved methods of immunization and control.

Effect of chronic trypanosomiasis on immunization against East Coast fever.

Two experiments were carried out in which uninfected cattle, or cattle chronically infected with Trypanosoma congolense, were immunized by the infection and treatment method against East Coast fever (ECF; Theileria parva infection). Chronic trypanosomiasis did not prevent cattle mounting an effective immunological response to ECF immunization and resisting subsequent lethal challenge. There appeared to be no difference in the level or quality of immunity between uninfected cattle and trypanosome-infected cattle. Thus, T. congolense infection on its own does not appear to provide a constraint to ECF immunization in the field.