Theileria parva: variation in the infection rate of the vector tick, Rhipicephalus appendiculatus

The variation in Theileria parva infection rates of experimental batches of adult Rhipicephalus appendiculatus ticks, used during the course of several years, was examined. It was found that considerable variation occurred, but that this could not always be correlated with the piroplasm parasitaemia in the cattle on which the ticks engorged as nymphs. Statistical analysis showed that the infection rate of ticks fed on cattle with a parasitaemia of 41–50 per cent was significantly higher than that of ticks fed on cattle with lower parasitaemias. A number of experiments were then carried out in which one or several factors of this aspect of the host-parasite relationship remained constant whilst others were altered. None of these factors was seen to play a major part in the variation. Finally, randomly selected groups of 10 ticks which had dropped engorged as nymphs from the same animal on the same day were examined. The variation observed even in these groups was so great that it was concluded that the infection rate could depend on a factor such as the juxtaposition of possibly-infected gut epithelial cells and developing salivary glands during the nymphal moult.     

East Coast Fever of Cattle: 60Co Irradiation of Infective Particles of Theileria parva

SYNOPSIS. Infective particles (IPs) of Theileria parva , the causative organism of East Coast Fever of cattle, were harvested from the tick vector, Rhipicephalus appendiculatus , using an in vitro feeding technic. In a ranging experiment, pairs of cattle were inoculated with aliquots of suspensions of IPs irradiated at doses of 4–137.6 krad. Doses of irradiation in excess of 8 krad appeared to destroy the parasite. In the 2nd and 3rd experiments, groups of 5 cattle were inoculated with aliquots of suspensions containing low and high concentrations of IPs respectively, irradiated at doses of 4–32 krad. In the 2nd experiment, doses of irradiation in excess of 10 krad appeared to destroy the parasite. In the 3rd experiment, at least 1 animal became infected when inoculated with an aliquot of a suspension irradiated at 16 krad. In all experiments, it appeared that increasing doses of irradiation destroyed increasing numbers of IPs. There was no conclusive evidence that IPs which survived irradiation were attenuated, and it appears that vaccination of cattle against ECF is unlikely to be achieved by inoculation of irradiated IPs using the methods described.     

Some factors controlling the stimulation of sporogony of Theileria parva in its tick vector Rhipicephalus appendiculatus

Young A. S., Leitch B. L. and Mutugi J. J. 1984. Some factors controlling the stimulation of sporogony of Theileria parva in its tick vector, Rhipicephalus appendiculatus. International Journal for Parasitology14: 97–102. The effect of various temperature treatments on the sporogony cycle of Theileria parva in the salivary glands of unfed adult Rhipicephalus appendiculatus ticks of various ages was investigated. It was found that ticks incubated at 28 or 37°C would develop sporozoites infective to cattle but never in such large numbers as in ticks fed on rabbits. Heat stimulation of sporogony was possible for isolates of T. parva with minimal laboratory handling. The age of the ticks incubated at 28 or 37°C was important since sporozoites could only be induced at the earliest on day 27 or 28 p.repl. (post-repletion) and at the latest by day 41 p.repl. The age of ticks fed on rabbits was not as important for the production of sporozoites.     

Development of resistance in rabbits to immature stages of the Ixodid tick Rhipicephalus appendiculatus

A study of acquired resistance in rabbits to larvae and nymphs of Rhipicephalus appendiculatus Neumann showed a positive correlation between the levels of gamma globulin in the serum and the resistance developed by the host. A negative correlation between the weight of engorged ticks and gamma globulin levels was also demonstrated during this study. Repeated infestations with ticks evoked a typical antibody response to antigenic challenge. The levels of gamma globulin stabilized after three infestations and no further decrease in the weight of ticks was observed. The mechanism of the acquisition of resistance by the rabbits, the production of antibodies and the involvement of complement are discussed.     

Salivary gland of the tick vector of East Coast fever. III. The ultrastructure of sporogony in Theileria parva

Sporogony of the sporozoan Theileria parva in the salivary gland of the tick vector of East Coast fever was studied in electron micrographs. The findings differ in several respects from previous interpretations based upon light microscopy. Cytokinesis of the primary sporoblast to form secondary and tertiary sporoblasts is not substantiated. Instead it is suggested that the parasite develops as a ramifying, multinucleate syncytium rapidly increasing in size and complexity until it gives rise to myriad sporozoites in a terminal episode of cytoplasmic fission. The proliferating nuclei initially occupy peripheral lobules that are continuous with a central labyrinth of branching and anastomosing processes which present a very large surface area for interchange of metabolites with the host cell cytoplasm. The membrane of the labyrinth is rich in cytostomes, but no evidence if found to bulk uptake of host cytoplasmic matrix or organelles into food vacuoles. Rhoptries are the first of the polar organelles of the parasite to develop and are associated with dense plaques irregularly distributed on the inner aspect of the parasite membrane. Micronemes form independently of the rhoptries at a later stage. After 3-4 days of tick feeding, sporogeny is complete and the infected salivary gland cell contains up to 50, 000 spherical or ovoid sporozoites about 1 micrometer in diameter. These are limited by a simple plasma membrane. The inner layer of the 'pellicle', the polar ring, and the conoid described for zoites of other Apicomplexa are lacking. Maturational changes are noted in sporozoites after sporogony is completed. Micronemes appear to increase in size, and possibly in number, from days 3-5 and the majority take up positions immediately subjacent to the plasmalemma.     

Genes transcribed in the salivary glands of female Rhipicephalus appendiculatus ticks infected with Theileria parva

We describe the generation of an auto-annotated index of genes that are expressed in the salivary glands of four-day fed female adult Rhipicephalus appendiculatus ticks. A total of 9162 EST sequences were derived from an uninfected tick cDNA library and 9844 ESTs were from a cDNA library from ticks infected with Theileria parva, which develop in type III salivary gland acini. There were no major differences between abundantly expressed ESTs from the two cDNA libraries, although there was evidence for an up-regulation in the expression of some glycine-rich proteins in infected salivary glands. Gene ontology terms were also assigned to sequences in the index and those with potential enzyme function were linked to the Kyoto encyclopedia of genes and genomes database, allowing reconstruction of metabolic pathways. Several genes code for previously characterized tick proteins such as receptors for myokinin or ecdysteroid and an immunosuppressive protein. cDNAs coding for homologs of heme-lipoproteins which are major components of tick hemolymph were identified by searching the database with published N-terminal peptide sequence data derived from biochemically purified Boophilus microplus proteins. The EST data will be a useful resource for construction of microarrays to probe vector biology, vector-host and vector-pathogen interactions and to underpin gene identification via proteomics approaches.     

East Coast fever: quantitative studies of Theileria parva in cattle

A series of experiments is described in which infective material obtained by grinding adult Rhipicephalus appendiculatus ticks containing mature Theileria parva parasites was titrated in East Coast fever-susceptible cattle. The reactions of the cattle to the various inocula, and the rate of multiplication of macroschizonts in their lymph nodes, were studied. In the final experiment, conclusive evidence was produced to support the observations of previous workers that the prepatent period, time to onset of febrile response, and time to death of the animals was dose-dependent, whereas the production of intraerythrocytic piroplasms was totally time-dependent. Furthermore, the effective rate of multiplication of macroschizonts was shown for the first time to be dose-dependent. It was not possible to detect macroschizonts before the fifth day after inoculation, and an occult phase of the parasite's life cycle, between the infective particle and the uninuclear macroschizont, is postulated. The discrepancies between the results of the present work and those of previous workers are discussed.     

Salivary gland of the tick vector of east coast fever. IV. Cell type selectivity and host cell responses to Theileria parva

Responses of cells in the tick salivary gland to parasitism by Theileria parva were studied by electron microscopy. The gland is composed of three distinct types of acini (I, II, III) which together include ten or more different cell types. Of some 30 infected cells observed in the present study, all were E-cells of acinus III. The parasite thus exhibits a high degree of selectivity for acinus and cell type. The glandular cell invaded undergoes massive hypertrophy and accumulates glycogen deposits in its cytoplasm which may serve as an energy source for the growing intracellular parasite. As synthesis of its secretory material declines the product is packaged in progressively smaller secretory granules. The extensive arrays of endoplasmic reticulum are dismantled and eliminated in autophagic vacuoles. Excess secretory granules are also broken down by crinophagy. After 4 days, sporogony is completed and the host cell contains 30,000–50,000 sporozoites in an electron-lucent cytoplasm largely devoid of cytomembranes and secretory granules. Mitochondria are still present and normal in appearance. The loss of basophilia and secretory granules observed heretofore by light microscopy have been attributed to ingestion and destruction of host organelles by the parasite. The pallid appearance of the cytoplasm has been interpreted as a sign of impending degeneration of the host cell. In electron micrographs no ingestion of organelles by the parasite or degenerative changes were found. The host cell clearly remains viable and metabolically active throughout sporogony. The striking changes in its ultrastructure result from active elimination of organelles and inclusions by the host cell itself in response to parasitism.     

Skin test to detect resistance of cattle to Rhipicephalus appendiculatus ticks

The evaluation of a skin test to detect acquired resistance to Rhipicephalus appendiculatus Neumann ticks is described. An extract of salivary glands of partially fed female R. appendiculatus was prepared by dissection, sonication and filtration and used for intra-dermal injection. Tests were carried out on twelve calves of known resistance to R. appendiculatus and on twelve naive calves to establish threshold values for positive reactions. Four rabbits naive to ticks were skin tested repeatedly to assess immunogenicity of the test. Reactions to the test at 1 h and at 24 h after injection were significantly correlated with resistance. The correlation was higher with the reactions at 24 h. Rabbits were immunized by the test but the reactions never exceeded the positive threshold. Further development by field testing is recommended.     

East Coast fever: the infectivity for cattle of infective particles of Theileria parva harvested in various substrates

Cunningham M.P., Brown C.G.D., Burridge M.J., Joyner L.P. and Purnell R.E. 1973. East Coast fever : the infectivity for cattle of infective particles of theileria parva harvested in various substrates. International Journal for Parasitology3: 335–338. Female Rhipicephalus appendiculatus ticks infected with Theileria parva were pre-fed on rabbits for 4 days before being removed and restrained on ‘plasticine’. They were then fed for 2-h periods on capillary tubes containing various substrates, the contents of which were subsequently inoculated into East Coast fever-susceptible cattle. Using this technique, precolostral calf serum and Eagle's Minimum Essential Medium, with the addition of Bovine Albumin Powder, were selected as substrates suitable for further laboratory investigation on East Coast fever as they were acceptable to the ticks and supported the viability of the parasites.     

Survival of Rhipicephalus appendiculatus (Acarina: Ixodidae) and persistence of Theileria parva (Apicomplexa: Theileriidae) in the field

Newson R. M., Chiera J. W., Young A. S., Dolan T. T., Cunningham M. P. and Radley D. D. 1984: Survival of Rhipicephalus appendiculatus (Acarina: Ixodidae) and persistence of Theileria parva (Apicomplexa: Theileriidae) in the field. International Journal for Parasitology14; 483–489. Two paddocks with populations of the African brown ear tick (Rhipicephalus appendicuiatus), one of which carried virulent Theileria parva (the causative organism of East Coast fever [ECF] of cattle), were left unstocked for periods of 338 and 354 days. Groups of 1–3, ECF-susceptible cattle were then introduced eight times during the following year to assess the tick and disease challenge, The ticks were also monitored continuously on the ground. The test cattle developed fatal ECF from adult ticks which had fasted for up to 554 days. A non-pathogenic, antigenicaliy distinct, Theileria species was also detected which was still transmitted by adult ticks after 600 days. The mean survival time of unfed larvae was 175 days (max. 240 days); unfed nymphs 270 and 450 (max. 540 days); unfed adults 420 days (max. 730 days), with females in a clear majority after 270 days.