East coast fever: Field challenge of cattle immunised againstTheileria parva (Muguga)

Summary This paper describes some of the work of the Edinburgh University East African Veterinary Expedition of 1969, carried out in conjunction with the F.A.O. Tickborne diseases project at Muguga. Cattle of proven immunity toT.parva (Muguga), together with controls, were exposed to field theirleriosis in two experiments. In both all animals died, though some protective effects of immunisation were demonstrable. The death of immunised animals is considered to be a result of aT.parva challenge by a strain of partial immunogenic difference fromT.parva (Muguga) concurrent with a profound anaemia caused byT.mutans infection.

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Sumario Este artículos describe parte del trabajo de la Expedición Veterinaria al Africa Oriental de la Universidad de Edimburgo en 1969 la cual, se realizó de acuerdo con el proyecto de “enfermedades transmitidas por garrapatas”, de FAO en Muguga. Ganado bovino de inmunidad probada haciaT. parva (Muguga), conjuntamente con controles, fueron expuestos a Theileriosis de campo en dos experimentos. Todos los animales de ambos grupos murieron, aunque se demostraron algunos efectos protectivos de inmunización. La muerte de los animales inmunizados se considera ser un resultado del desafió conT. parva con una cepa de una differencia inmunogénica parcial deT. parva de Muguga, concurrente con una profunda anemia causada por la infección conT. mutans.

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Résumé L’article décrit une partie du travail de l’expédition vétérinaire de l’Université d’Edinburgh, réalisé en Afrique de l’Est en 1969, en collaboration avec le projet de la FAO pour les maladies transmises par les tiques à Muguga. Des bovins dont l’immunité àT. parva (Muguga) avait été prouvée, ont été exposés, avec des témoins, à la theilériose naturelle au cours de deux expériences. Dans les deux cas, tous les animaux sont morts, bien qu’un certain effet protecteur de l’immunisation ait pu être démontré. On considère que la mort des animaux immunisés est due à l’infection par une souche deT. parva ayant une structure immunogène partiellement différent de celle deT. parva (Muguga), combinée à une anémie intense causée par une infection parT. mutans.

     

East coast fever: 4. Further studies on the protection of cattle immunized with a combination of theilerial strains

Four experiments were carried out in which cattle immunized with three cryopreserved stabilates, Theileria parva (Muguga), T. parva (Kiambu 5) and T. lawrencei (Serengeti transformed) and simultaneously treated with a single dose of long acting tetracycline were challenged with field isolates of T. parva. Three of the challenge isolates were from Tanzania and one from Kenya. The immunized cattle showed either mild or inapparent diease reations while susceptible control cattle suffered severe disease and 72% died. These experiments further demonstrated the efficacy of the immunization method against T. parva isolates from a wide distribution within East Africa. The value of such laboratory screening of field isolates prior to field exposures is discussed.     

East coast fever: Cultivationin vitro of bovine spleen cell lines infected and transformed byTheileria parva

Summary Three spleen cell lines derived from 3 calves experimentally infected withTheileria parva have been established. After an apparent disappearance of parasitized cells from the cultures there was a resurgence of infected lymphoblasts in 5 to 6 weeks. The infected lymphoblasts can be readily subcultured without feeder layers in a medium consisting of Eagle’s Minimal Essential Medium and 20 per cent foetal calf serum. The plating of infected cells was greatly enhanced by implanting the cells on preformed monolayers. No evidence was obtained by the methods employed which might indicate the existence of an extracellular form of the parasite capable of reinfecting new cellsin vitro.

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Sumario Se han establecido 3 lineas celulares de bazo derivadas de tres terneros experimentalmente infectados conTheileria parva. Despuéś de una aparente desaparición de las células parasitadas de los cultivos, hubo una resurgencia de linfoblastos infectados en 5–6 semanas. Los linfoblastos infectados pueden ser subcultivados facilmente sin capas alimentadoras en un medio consistente de Eagles, medio mínimo esencial enriquecido con 20 por ciento de suero bovino fetal. El plaqueo de las células infectadas fue grandemente estimulado mediante la implantación de das células en monocapas previamente formadas. Con los métodos utilizados no se obtuvo evidencia para indicar la existencia de una forma extra celular de el parásito capaz de reinfectar neuvas célulasen vitro.

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Résumé Trois lignées de cellules de rate issues de trois veaux expérimentalement infectés parTheileria parva ont été établies. Après une apparente disparition des cellules parasitées dans les cultures, il y avait une résurgence de lymphoblastes infectés au bout de 5 à 6 semaines. Ceux-ci pouvaient être mis en subculture aisément sans couche de soutien dans un milieu constitué du M.E.M. de Eagle, enrichi par 20 p. 100 de sérum de foetus de veau. L’étalement des cellules infectées était grandement facilité par leur implantation sur des couches préformées. Par les méthodes employées, on obtient aucune preuve de l’existence d’une forme extracellulaire du parasite, capable de réinfecterin vitro des cellules neuves.

     

Kinetics of infection with Theileria parva (East Coast fever) in the central lymph of cattle

During the course of a lethal infection with Theileria parva in susceptible cattle, the dissemination of the parasite was examined in central lymph efferent from superficial lymph nodes in the thoracic duct. From the regional node, lymphocytes containing macroschizonts of T. parva were detected in efferent lymph 8 days after challenge where their appearance coincided with a dramatic increase in the output of lymphoblasts. The number of infected cells reached a maximum around Day 14, when 60-65% of efferent lymphocytes were parasitized. A severe reduction in the total cell output occurred after Day 14, at the time when widespread lymphocytosis was observed in the parent lymph node. A similar pattern of cellular kinetics was observed in the thoracic duct and in lymph efferent from lymph nodes distant from the site of challenge, although in the latter, the parasitosis reached only 10% of total cells. There was no selective depletion of parasitized cells from central lymph during the third week of infection, although the comparative parasitosis between lymph and lymph node cells indicated that infected cells entered central lymph less readily during this period. Macroschizonts appeared in cultures of lymphatic lymphocytes sampled between 5-9 days after challenge. These results, together with the failure of ablation of the regional lymph node 2, 3 or 5 days after challenge to delay the onset of the disease, indicated that dissemination of the infection from the site of challenge occurred within the first 2-3 days after the inoculation T. parva.     

Chemotherapy of East Coast fever: parvaquone treatment of clinical disease induced by isolates of Theileria parva

Two experiments were carried out in which parvaquone was used to treat experimentally-induced acute clinical East Coast fever infections. In the first experiment, infections with Theileria parva parva (Kiambu 5) were induced by applying infected Rhipicephalus appendiculatus ticks or by inoculation of triturated infected-tick stabilate. The character of the disease was similar with both methods of infection and following a single treatment with parvaquone at 20 mg kg-1, 5 of 7 cattle in each group recovered. All untreated control cattle died. In the second experiment, 5 stabilate isolates from different locations within East Africa, and representative of the challenge likely to be met in the field, were used. Treatment was administered in 2 X 10 mg kg-1 doses 48 h apart. The isolates used were T. p. parva (Mbita), T. p. parva (Pugu), T. p. parva (Entebbe), T. p. lawrencei (Mara) and T. p. lawrencei/(Manyara); following treatment 3/7, 6/6, 6/7, 5/7 and 6/7 animals recovered, respectively. All untreated control cattle died. There was evidence of a difference in susceptibility of isolates to treatment, and some animals showed prolonged disease episodes. The nature of the response to treatment and the problems in treating a lympho-destructive disease are discussed.     

Clinical and diagnostic features of East Coast fever (Theileria parva) infection of cattle

East Coast fever is a tick-borne protozoal disease affecting cattle in a large part of East and Central Africa. Since the vector occurs over an even wider range there is considerable potential for the disease to spread to countries which are currently disease free. This article, describing the clinical and diagnostic features of East Coast fever, may remind authorities in these countries of the potential hazards posed by the disease.     

East coast fever: 2. Cross-immunity trials with a Kenya strain of Theileria lawrencei

Cross-immunity trials were carried out on cattle which were immunized against theileriosis either by chemoprophylaxis or by sub-lethal challenge. In the first of two experiments, animals were immunized against either one of two strains of Theileria parva or one of two strains of Theileria lawrencei. They were then challenged with a Kenya strain of T. lawrencei (T. lawrencei KB 5). The former animals were not protected against the challenge, whereas the latter were either partially protected, when a Tanzanian strain of T. lawrencei had been used for immunization, or completely protected when another Kenyan strain of T. lawrencei had been used.In the second experiment, animals were immunized by chemoprophylaxis against T. lawrencei (KB5) and challenged with T. parva (one strain) or T. lawrencei (five strains). The cattle were protected against challenge with the identical or related T. lawrencei strains, but only partially protected against the T. parva strain and a Tanzanian strain of T. lawrencei.It appears that cattle cannot be immunized for field exposure to theileriosis throughout East Africa by chemoprophylaxis using T. lawrencei (KB5) alone, but may be able to withstand field exposure in a variety of situations if a combination of different theilerial strains is used in the vaccination procedure.     

Chemotherapy of East Coast fever: parvaquone treatment of Theileria parva parva at intervals after infection

Groups of seven cattle were infected with Theileria parva stabilate and treated with parvaquone (20 mg kg-1 bodyweight) zero, four, eight, 12, 14 or 16 days after infection. Very early treatments resulted in a rapid recovery or no detectable parasitosis and some cattle were subsequently susceptible to homologous challenge. Treatments applied before extensive lymphoid or other organ damage had developed were successful and some cattle treated in advanced disease also recovered. Clinical pathological indications of liver or kidney damage were recorded very late in the disease suggesting that prompt diagnosis and treatment are more important than supportive therapy for survival. Treatment on day 8 after infection allowed the appearance of macroschizonts and a transient pancytopenia but no other disease signs. This group was solidly immune to challenge and this timing and treatment could be recommended for use in an infection and treatment method of immunisation.     

Induction of immunity against infection with Theileria parva (East Coast fever) in cattle using plasma membranes from parasitized lymphoblasts

Protection against challenge with Theileria parva was conferred on three of four calves given three or four inocula of plasma membranes prepared from 6 to 12 X 10(8) autologous parasitized lymphoblasts from cultured cell lines. In contrast, calves remained susceptible to infection following immunization with membranes prepared from allogeneic parasitized lymphoblasts. Similarly, calves vaccinated with either gamma-irradiated autologous or allogeneic infected cells also died of East Coast fever after challenge. The results raise the possibility of vaccination against T. parva using subcellular material from infected lymphoblasts.     

East Coast fever immunisation field trial in crossbred dairy cattle in Hanang and Handeni districts in northern Tanzania

East Coast fever (ECF) causes considerable mortality and production losses in the Tanzania smallholder dairy sector and limits the introduction of improved dairy breeds in areas where the disease is present. The infection and treatment method (ITM) was adopted by smallholder dairy farms for ECF immunisation in Hanang and Handeni districts of Tanzania. This study recorded incidence rates for ECF and other tick-borne diseases (TBDs) for ECF-immunised and non-immunised cattle between 1997 and 2000. Approximately 80% of smallholder households from both sites (n?=?167) participated in this longitudinal study, with immunisations carried out at the request of the livestock owners. Efficacy of ITM for preventing ECF cases in these crossbred dairy cattle was estimated at 97.6%, while that for preventing ECF deaths was 97.9%. One percent of the cattle developed clinical ECF as a result of immunisation. Since ECF immunisation permits a reduction in acaricide use, an increase in other TBDs is a potential concern. Sixty-three percent of farmers continued to use the same acaricide after immunisation, with 80% of these reducing the frequency of applications. Overall, 78% of farmers increased the acaricide application interval after immunisation beyond that recommended by the manufacturer, resulting in annual savings in the region of USD 4.77 per animal. No statistical difference was observed between the immunised and non-immunised animals in the incidence of non-ECF TBDs. However, immunised animals that succumbed to these diseases showed fewer case fatalities. ITM would therefore appear to be a suitable method for ECF control in Tanzania's smallholder dairy sector.     

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)     

East coast fever: 3. Chemoprophylactic immunization of cattle using oxytetracycline and a combination of theilerial strains

In the first of two experiments performed, 29 animals were immunized by chemoprophylaxis against either a combination of three theilerial strains, Theileria parva (Muguga), T. parva (Kiambu 5) and T. lawrencei (Serengeti transformed), or against one or two elements of the combination separately, and then challenged with either T. parva (Kiambu 1) or T. lawrencei (Solio KB1), both of which were known to be heterologous to T. parva (Muguga). The animals immunized against the combination had mild or inapparent reactions to challenge, whilst the susceptible control cattle and many of those immunized against one or two strains either had severe reactions of died.In the second experiment, 17 cattle immunized by chemoprophylaxis against the combination were challenged with lethal doses of three theilerial strains, T. parva (Entebbe 1), T. parva (Entebbe 2) and T. parva (Ukunda), recently isolated from two widely separated areas of East Africa. They had mild or inapparent reactions whilst 13 of 15 susceptible control cattle died.Chemoprophylaxis using a combination of theilerial strains and a limited oxytetracycline regimen was shown to be an effective means of immunizing cattle against East African theilerioses. The use of this technique as a method of immunizing cattle before exposure in the field is proposed.     

Clinical efficacy and plasma concentrations of two formulations of buparvaquone in cattle infected with East Coast fever (Theileria parva infection)

East Coast fever, caused by the protozoan parasite Theileria parva, kills about 600,000 cattle annually in Africa. The hydroxynaphthoquinone compound buparvaquone (BPQ) is curative. Sixteen calves were infected with T. parva. On manifestation of disease symptoms, eight were injected with the original (pioneer) BPQ product and eight with a test product containing BPQ. All 16 calves were cured by one injection of 2.5 mg BPQ/kg bodyweight. The concentration of BPQ in blood plasma was monitored by HPLC. The mean observed C(max) of BPQ was 0.229 and 0.253 microg/mL of plasma, the mean observed time to reach this concentration (T(max)) was 2.62 and 2.12 h and the AUC (area under curve) was 4.785 and 4.156 microg h/mL, respectively, for the pioneer and test product. Considerable variations occurred in the plasma concentration of BPQ within each group. They showed no relationship with either clinical or parasitological parameters following treatment.     

Tick burden and prevalence of Theileria parva infection in Tarime zebu cattle in the lake zone of Tanzania

This study was carried out to assess the distribution, abundance of different tick genera and prevalence of Theileria parva infection in Tarime zebu cattle kept in selected wards of Serengeti and Tarime districts in Mara region. Adult ticks were identified and counted from half body parts of 360 animals which were extensively managed in communal land with natural pastures. Concurrently, blood samples were collected and thereafter DNA extracted and a nested polymerase chain reaction (nPCR) was done using primers specific for p104 gene to detect the presence of T. parva DNA. Ticks were identified into four groups: Amblyomma genus, Boophilus sub-genus of Rhipicephalus genus, other species of Rhipicephalus, and Hyalomma genus. Rhipicephalus genus accounted for 71.8 % of the total ticks, whereas Amblyomma, Boophilus sub-genus of Rhipicephalus genus and Hyalomma constituted 14.1, 14.0 and 0.1 %, respectively. There were more animals (p?p?T. parva was 27.7 % and was higher (p?T. parva did not show any clinical signs of East Coast fever (ECF), suggesting the existence of subclinical infection in Tarime zebu. These results suggest that Tarime cattle can tolerate ECF infection and are likely to serve as potential carriers of T. parva to other less-tolerant cattle breeds in mixed herds. Since Tarime cattle are preferred by most farmers with mixed herds, routine screening for T. parva is highly recommended to minimize introduction of infected cattle into an immunologically naive population.     

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.