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The paper reviews the infectivity, cross-immunization experiments, and cattle vaccination of Zimbabwean cattle-derived Theileria parva (Boleni) sporozoite stabilates produced at the Central Veterinary Laboratory (CVL) in Harare between 1980 and 2003. The Boleni stock was first isolated in July 1978 during a theileriosis outbreak and was shown to be virulent in susceptible cattle. Thereafter, the reactions observed in susceptible cattle produced by different tick stabilates derived from the original have been mostly severe (76%) or moderate (24%). The parasite concentrations in the Boleni vaccine, the vaccines used in East Africa, and a Malawian stock were compared. The infective Theileria sporozoite concentration in 1 ml of stabilate in the Muguga and Serengeti (from East Africa) and Kasoba (from Malawi) vaccines were 8×, 9×, and 14× the concentration of the Boleni stabilate, respectively. The Boleni strain, like the other Zimbabwean T. parva isolates, produces a characteristic low piroplasm parasitaemia of usually less than 1% in susceptible cattle. This has largely contributed to the difference in infection rates (1963; average 40%) among tick batches used to prepare the various stabilates. Subsequently, the sporozoite concentrations in 1 ml of stabilate also varied considerably (6-91; average 53), making the reproducibility and standardization of the stabilates for immunization difficult. Immunization of cattle using Boleni stabilates with oxytetracycline therapy or with titrated low doses without treatment was found to be safe and efficacious. Cross-immunity experiments demonstrated that T. parva Boleni stabilates cross-protected against all the Zimbabwean cattle-derived T. parva stocks tested. The characteristics of the Boleni stock in affording a wide spectrum of cross-protection make it an excellent candidate for cattle immunization in Zimbabwe, hence protecting the country from the introduction of foreign vaccines and subsequently, foreign parasite populations., (Copyright © 2011 Elsevier GmbH. All rights reserved.)

The purpose of this study was to determine the species spectrum of ticks infesting goats owned by resource-limited farmers in the state-owned communal land areas of Zimbabwe. Ticks were collected from goats at a single locality within each of 5 communal land areas, and a total of 14 ixodid tick species was recovered. The most numerous tick was Rhipicephalus evertsi evertsi, which was present in all areas at each sampling, and a Rhipicephalus sp. (near R. punctatus), which was most abundant on goats in the central regions of Zimbabwe during the March rainy season. Amblyomma hebraeum was present on goats in all areas sampled. In the eastern central region its distribution overlapped that of Amblyomma variegatum, while in the northwest it overlapped those of both Amblyomma marmoreum and A. variegatum. Hyalomma truncatum was present at all localities, whereas only a single Hyalomma rufipes was recovered. Rhipicephalus appendiculatus was collected from goats in the moist, slightly cooler regions, while the few Rhipicephalus zambeziensis recovered were present in the hotter, drier regions. Species recorded in lower numbers were Rhipicephalus (Boophilus) decoloratus, Rhipicephalus lunulatus, Rhipicephalus simus, Rhipicephalus tricuspis and Rhipicephalus turanicus. Attachment in the inter-digital space of adult A. hebreaum and H. truncatum was sometimes associated with lameness.

There has been an increase in the introduction of game animals, including African buffaloes, into the Highveld of Zimbabwe to establish private game reserves on condition that they are confined in separate and secured paddocks. Owing to shortages of pastures cattle were grazed in buffalo-grazed paddocks resulting in outbreaks of buffalo-derived theileriosis. This paper reports the results of epidemiological observations carried out on two game reserves to assess the risk of buffalo translocation. The infection rate with Theileria parasites in ticks collected from buffalo-grazed pastures was high and produced fatal theileriosis in susceptible cattle. Similarly, adult R. appendiculatus ticks artificially fed as nymphs on the buffaloes produced fatal infections in susceptible cattle. Theileria parva (Boleni), the vaccine used to immunize cattle against theileriosis, and a buffalo-derived T. parva stabilate (BV-1) were inoculated in naïve buffaloes to study the Theileria carrier-state in these animals. The two buffaloes that had received the Boleni stabilate showed no clinical theileriosis reaction; however, the ticks derived from them produced a subclinical reaction in one susceptible calf. The buffalo which had received stabilate BV-1 developed fever, high schizont parasitosis for 10 days and 15% piroplasms parasitemia. R. appendiculatus ticks fed as nymphs on this buffalo produced fatal theileriosis reaction in a susceptible calf.

The Theileria parva carrier-state in cattle on commercial farms on Zimbabwe was investigated using parasitological and serological methods. The proportion of cattle showing Theileria piroplasms on two farms, which had recent histories of disease outbreaks, were 64% (n = 106, total of heifers and weaned calves examined) and 71.5% (n = 60) while the proportion of T. parva antibodies for the same animals were 59% and 98.5%, respectively. On four farms where no cases of the disease occurred for over 10 years, the average proportion of animals showing piroplasms and antibodies were 55.4% (range 32-82, n = 223) and 73% (range 47-91, n = 223), respectively. However, on another three farms which had no history of theileriosis outbreaks these proportions were very low, being 11.4% (0-24, n = 157) for piroplasms and 12.2% (5-23, n = 157) for antibodies. The mean infection rate in unfed Rhipicephalus appendiculatus adults collected from farms with a high prevalence of cattle which were carriers of Theileria piroplasms during the tick activity season was 29% (range 12-60%) with 9.3 (range 2-18.7) mean infected acini per infected tick. The infectivity of different tick batches to susceptible cattle produced a wide spectrum of theileriosis reactions. Laboratory controlled experiments were carried out to study the persistence of T. parva (Boleni) piroplasms in cattle immunized with this strain as well as its infectivity for ticks and its subsequent transmissibility to cattle. Examination of the salivary glands of 15 batches of ticks collected from six immunized cattle on three different occasions over 18 months showed that none were infected with Theileria parasites. However, the infectivity of other ticks in the same batches to susceptible animals was demonstrated 6, 10 and 18 months after cattle had been immunized with Boleni stabilate.

Fifty-nine Hereford cattle susceptible to tick-borne diseases were used as tracer animals to assess the tick challenge and pathogenicity of Theileria parva under field conditions in Zimbabwe. They were moved periodically in groups of five to three commercial farms (one group consisted of four) during seasons of Rhipicephalus appendiculatus nymphal and adult activity. All tracer cattle were herded together with the farm cattle but were not dipped. The nymphal tick counts were high on two of the farms (up to 2000 per animal) but were very low on the third farm (less than ten per animal). On the three farms, 19 out of 24 (76%) tracers had patent Theileria schizonts. There was a range of clinical manifestations of theileriosis with acute and fatal infections occurring on one farm. The adult R. appendiculatus infestations during the wet season numbered 120-800 per animal on the three farms. The disease transmitted by the adults was very pathogenic on the three farms; 30 out of 35 (86%) had severe theileriosis infections. Cattle, which survived the nymphal diseases challenge, showed various degrees of immunity to subsequent T. parva challenge transmitted by adult ticks. Therefore, 13 out of 18 (72%) of these cattle had a second disease episode and the case fatality rate on the three farms was 46%. The factors which determined the epidemiological status of Theileria challenge on the farms, such as the farming systems and presence of wild animals, are discussed.

Eight cattle immunized with cattle-derived Theileria parva Boleni stabilate together with six susceptible controls were released in Dombawera Game Park on the Highveld of Zimbabwe. This coincided with Rhipicephalus appendiculatus nymphal activity. The cattle grazed together with African buffaloes (Syncerus caffer) and were not treated against tick infestation. The nymphal tick infestation was high, and seven of the eight immunized cattle and three of the controls had severe and fatal reactions. Subsequently, two stocks of Theileria parva to be tested for their immunizing abilities were prepared-one from adult ticks which were fed as nymphs on one of the sick control animals (Dom 268) and the other from adult ticks collected from pastures grazed by buffaloes (Bv-1). Two groups of cattle were immunized with either the Dom 268-derived strain (eight animals) or the Bv-1-derived strain (four animals). These together with three non-immunized controls, were released in Bally Vaughaun Game Park in the Highveld, where buffaloes are present, during the season of nymphal tick activity. A third group of five cattle, immunized with stabilate Bv-1, and three non-immunized controls were released at the same site during the season of adult tick activity. The nymphal and adult tick infestations of the cattle were large and more than 2000 nymphs and 1000 adult ticks were counted per animal. Cattle were treated with a pyrethroid pour-on preparation to control the tick infestation and screw-worm strike. The immunized cattle in the three groups survived the theileriosis challenge for a period of 18 months, but the non-immunized control cattle suffered a severe and fatal theileriosis 19-23 days after being placed on the pasture.

Over 500 cattle of all age groups on 2 farms in Zimbabwe were immunised against theileriosis using the "infection and treatment" method and disease prevalence and their productivity assessed during a period of 18 months. The immunising stock, Theileria parva (Boleni) was isolated in Zimbabwe. None of the immunised cattle suffered from theileriosis upon natural exposure whereas 22 unimmunised cattle died of theileriosis and a further 48 required treatment for theileriosis. In the first year, some immunised cattle were maintained with minimal threshold dipping (once or twice during the rainy season). During periods of very high tick challenge of 100 to 1,000 Rhipicephalus appendiculatus per animal from January to March, a transient decrease in liveweight gain was observed particularly in cows. However, by the end of the period of observation, the weights had recovered so that intensively dipped and immunised and threshold or strategically dipped groups of cattle showed no significant differences. From the results it was estimated that each engorging female R. appendiculatus caused a temporary depression in weight gain of 8 grams. In young stock the weight loss was exacerbated by the presence of screw worm (Chrysomya bezziana) infestation. It was then possible to define an economically attractive integrated tick and theileriosis control strategy based on these findings, whereby immunised cattle were dipped 6 times between mid-December and mid-March. In this regimen, no weight loss occurred and no cases of screw worm were observed. For each of 3 herd sizes of 250, 500 and 1,000 cattle, comparisons were made of the costs of 4 different control options: (i) Intensive dipping (40 times/year) (ii) Intensive pour-on acaricide treatments (18 times/year) (iii) Theileria immunisation with strategic dipping (6 times/year) (iv) Theileria immunisation with pour-on treatment (4 times/year) It is concluded from these studies that, on farms where theileriosis is a serious problem, immunisation coupled with a strategic dipping programme is economically very attractive. In the year in which immunisation is carried out, costs will be higher than for intensive dipping, but from the second year on, the costs are decreased to approximately 50% of those for intensive dipping.

Forty Theileria schizont-infected lymphocyte culture isolates from Zimbabwe were characterized using a panel of antischizont monoclonal antibodies (MAbs) and 4 Theileria parva DNA probes containing cloned extrachromosomal element, Tpr repetitive, ribosomal and telomeric sequences. The Theileria isolates were assigned as T. parva or T. taurotragi on the basis of reactivities with MAbs and restriction fragment length polymorphisms (RFLPs) detected using the extra-chromosomal element probe. Cattle-derived T. parva isolates were relatively homogeneous on the basis of reactivities with MAbs and RFLPs detected using Tpr repetitive and ribosomal DNA probes. In contrast to previous results from Kenya, most of the cattle-derived isolates from Zimbabwe exhibited very similar Tpr restriction fragment patterns, although the Tpr genotypes of buffalo-derived isolates were heterogeneous. This suggests that selection for a particular Tpr genotype may be occurring in cattle. Many isolates with similar Tpr genotypes were differentiated by RFLPs detected using the telomeric DNA probe. The T. parva Boleni immunizing stock was distinguished from all other isolates by telomeric RFLPs. The T. parva Boleni Tpr repetitive DNA probe cross-hybridized with T. taurotragi DNA and detected RFLPs between different T. taurotragi isolates.