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PubMed1981

The initial stages of infection of cattle cells with Theileria parva sporozoites in vitro.

Large numbers of Theileria parva sporozoites were separated from Rhipicephalus appendiculatus adult ticks by filtration and were then concentrated by centrifugation. The sporozoites were incubated at 37 degrees C with leucocytes from 6 cattle of Bos indicus and B. taurus types. Giemsa-stained smears and living preparations under interference contrast microscopy were used to follow the course of the infection of the leucocytes with sporozoites. Sporozoites were seen to attach rapidly to about 25% of the leucocytes which they penetrated. After penetration by the sporozoites the morphology of the cells changed to show an increase in cytoplasm and an enlargement of the Golgi apparatus, with which the parasite appeared to become associated. The early intracellular or preschizont stages resembled Babesia parasites. From day 3, the parasite showed the typical morphology of the macroschizont of T. parva. Multiple infections were frequent and up to 8 schizonts were observed arranged around the Golgi apparatus. Multiple infected cells did not survive in culture but some of the cells infected with a single parasite divided to produce 2 infected daughter cells and infected lymphoblastoid cell lines were established in all 21 attempts.

Animals↗
PubMed1981

Epidemiology of East Coast fever: some effects of temperature on the development of Theileria parva in the tick vector, Rhipicephalus appendiculatus.

The moulting behaviour of nymphal Rhipicephalus appendiculatus from the laboratory colony and 3 field strains from Kenya was greatly influenced by constant temperatures between 18 and 37 degrees C but was not significantly different for the 4 tick strains. Six batches of R. appendiculatus (Muguga laboratory) nymphs which had engorged on cattle parasitaemic with 4 stocks of Theileria parva (Kiambu 4, Kiambu 5, Muguga and Mbita 4) isolated in Kenya were incubated at constant temperatures between 18 and 37 degrees C during their pre-moult and post-moult periods. The Theileria infections in the salivary glands of the resultant adult ticks were assessed by staining with Feulgen's stain. Two stocks (Muguga and Kiambu 5) developed their highest infections in ticks incubated at 28 degrees C while the other two (Kiambu 4 and Mbita 4) developed their highest infections at 23 degrees C. Constant temperatures of 18, 33 and 37 degrees were detrimental to the development of salivary gland infections. Temperature influenced the rate of development and numbers of the earlier Theileria stages in the ticks. Engorged nymphal ticks incubated for 4 days at 37 degrees C during their pre-moult period developed lower infections than ticks exposed at 4 degrees C for 4 days.

Animals↗
PubMed1978

Development of infectivity in Hyalomma detritum (Schulze, 1919) ticks infected with Theileria annulata (Dchunkowsky and Luhs, 1904).

The time-course for the development of infectivity was studied in Hyalomma detritum ticks fed as pre-imagoes on calves infected with Theileria annulata. Unfed adults derived from infected nymphs were non-infectious when inoculated into susceptible calves, whereas ticks of both sexes that had fed for 2-3 days or longer on calves or rabbits were always infectious. Some adults fed for only 1-2 days were infectious, whereas others were not. As few as 2 ticks were capable of infecting susceptible calves.

Animals↗
PubMed1983

Infection of mammalian cells with Theileria species.

Experiments were carried out to determine the susceptibility of mammalian cells to infection with different species of Theileria in vitro. Sporozoites of Theileria parva (parva), Theileria parva (lawrencei) and Theileria taurotragi were isolated from Rhipicephalus appendiculatus ticks by grinding infected ticks in medium, filtering the suspension and concentrating by centrifugation. The sporozoites were used in attempts to infect in vitro peripheral blood leucocytes harvested from 16 different mammalian species which included 12 species of Bovidae from 6 different sub-families. The technique was shown to be both sensitive and reproducible. The sporozoites of T. parva (parva) infected and transformed cells from 2 species of the sub-family Bovinae, the two cattle types and African buffalo. Theileria parva (lawrencei) infected and transformed cells from the two cattle types, African buffalo and Defassa waterbuck. Theileria taurotragi sporozoites infected in vitro cells from 11 different species of Bovidae which were members of 6 sub-families; Bovinae, Tragelaphinae, Reduncinae, Alcelaphinae, Antilopinae and Caprinae. Transformed lymphoblastoid cell lines were established from 7 of the species infected. Sporozoite attachment and infection was not observed with non-susceptible bovid host cells, nor were any of the non-bovid leucocytes infected by the parasites. The host range observed in this study corresponded to the known host range in vivo.

Animals↗
PubMed1983

Transmission of Theileria parva by a population of Rhipicephalus appendiculatus under simulated natural conditions.

A 2 hectare paddock on the Muguga Estate, Kiambu District of Kenya (altitude 2100 m) free of Theileria parva-infected ticks was seeded by applying Rhipicephalus appendiculatus nymphs to cattle infected with Theileria parva (Kiambu 4). It was estimated that over 50 000 engorged nymphs fell onto the pasture from 4 cattle with high parasitaemias during the cold season (June). Samples of these ticks were placed in plastic cylinders under the vegetation in the paddock and sexual stages of T. parva were detected in gut lumen smears in these samples up to 21 days after repletion. Zygotes were first observed to transform into kinetes on day 55 and parasites were first detected in salivary glands of adults on day 64. Moulting of the nymphs started on day 60 after repletion and was completed by day 87. Cattle introduced into the paddock showed their first infestation with adult R. appendiculatus on day 64 after repletion of the nymphs and the infestation level gradually increased. On day 76 after repletion, 17% of a sample of adult ticks infesting cattle showed salivary gland infections with T. parva and this increased to over 70% by day 150. From day 360 onwards, a decrease in both the percentage of ticks infected and the number of salivary gland acini infected/tick was noted. In addition, T. parva infections within the salivary glands required a longer period of feeding in the older ticks before they developed into sporozoites. Cycling of Theileria through ticks was prevented by the removal of female ticks before they completed repletion. Lethal challenge levels of T. parva for cattle existed in the paddock up to day 547 after repletion, after which introduced cattle showed no infection or only a sub-lethal infection. Cattle introduced into the paddock on day 808 after repletion showed no evidence of T. parva infection. The tick infestation of introduced cattle decreased markedly from day 368 and infestation had virtually ceased by day 808.

Animals↗
PubMed1983

Transmission of Theileria annulata (Dschunkowsky & Luhs, 1904) by Hyalomma excavatum (Koch, 1844).

Theileria annulata was transmitted transtadially by an Israeli strain of Hyalomma excavatum from larvae to nymphs and from nymphs to adults. However, the adults developing from larvae which engorged on infected calves and fed during the nymphal stage on non-infected calves, rabbits or gerbils (Meriones tristrami), did not transmit theileriosis. Suspensions of infected unfed nymphs or adults did not produce theileriosis but both nymphs and adults of the same batch became infective to cattle after 2 days of feeding. Suspensions of salivary glands or of whole ticks derived from infected adults that remained attached to rabbits for more than 7 days, were not infective to cattle.

Animals↗
PubMed1975

Correlation between the morphology and infectivity of Theileria lawrencei developing in the tick Rhipicephalus appendiculatus.

Adult Rhipicephalus appendiculatus ticks, infected as nymphs by feeding them on an African buffalo harbouring Theileria lawrencei, were applied to the ears of rabbits. Equal numbers of the feeding ticks were removed daily from rabbits and either dissected and processed so that parasites in their salivary glands could be examined morphologically and classified, or ground in Eagle's Minimal Essential Medium, to harvest parasites from the ticks in the resultant supernatant fluid. The infectivity of the supernatant fluid was tested by inoculation of aliquots into groups of susceptible cattle. It was found that supernatant fluids produced from day 0 to day 2 fed ticks were uninfective to cattle but those produced from 3 to 9 days were infective. The most infective supernatant fluid was produced from 6 day fed ticks. The reactions resulting in cattle were correlated with the number and morphology of parasites in sections of salivary glands and it was found that mature parasites coincided with infectivity of the supernatant fluid to cattle. From the reactions in cattle inoculated with supernatant fluids from 5 and 6 day fed ticks it was concluded that this method of harvesting parasites was efficient. Examination of the salivary glands of a representative sample of a T. lawrencei infected tick batch is a potential means of screening material for T. lawrencei stabilates.

Animals↗
PubMed1987

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.

Animals↗
PubMed1987

An enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to Encephalitozoon cuniculi and its use in determination of infections in man.

An enzyme-linked immunosorbent assay (ELISA) was used to detect antibodies to Encephalitozoon cuniculi in man, using spores derived from tissue culture as antigen. Negligible cross-reactions were found with other microsporidia of vertebrate or of invertebrate origin and there was no cross-reaction with Toxoplasma gondii, using ELISA, immunoperoxidase or immunofluorescence staining. A high prevalence of antibodies to E. cuniculi was found in patients suffering from schistosomiasis, malaria and neurological and psychiatric disorders, but not in healthy individuals.

Animals↗
PubMed1984

Determination of nuclear DNA of five eucoccidian parasites, Isospora (Toxoplasma) gondii, Sarcocystis cruzi, Eimeria tenella, E. acervulina and Plasmodium berghei, with special reference to gamontogenesis and meiosis in I. (T.) gondii.

DNA contents of individual stages of Isospora (Toxoplasma) gondii and other Eucoccida were measured after Feulgen-pararosaniline (SO2) staining either by direct microfluorometry or by scanning of microphotographic negatives. Frequency distributions were analysed using a computer program based on a mathematical model describing cell division. All stages of I. (T.) gondii, except fertilized macrogametes (2c), contained a haploid amount of DNA (1c), indicating that meiosis in I. (T.) gondii occurs during sporogony. Microgametes possessed 3.3% DNA in excess, presumably mitochondrial DNA. Nuclei of M2- and M3-merozoites differed in two characteristics: a small but distinct nucleolus was observed in almost 50% of the M2-merozoites but in none of the M3-merozoites; all M2 merozoites were strictly haploid, while all M3-merozoites were synthesizing DNA (17% above the haploid value). It may be concluded that all M2- and M3-merozoites are already sexually differentiated, i.e. are macro- and microgamontoblasts, respectively. DNA synthesis necessary for the development of the microgamont starts already in the microgamontoblast stage (M3-merozoite). M2-merozoites macrogametes, synthesize 11% extra DNA before fertilization, (after fertilization an extra amount of 12% of the diploid value was found), probably by amplification of genes for proteins which are needed for rapid maturation and later sporogony. Essentially parallel results have been found in Eimeria tenella and in crescent cystozoites of Sarcocystis cruzi. Absolute DNA values in representatives of the Eucoccida have been estimated as follows (10(-15) g): I. (T.) gondii, 96; E. tenella and E. acervulina, both 75; S. cruzi, 216; Plasmodium berghei, 27. The value of the estimation of total haploid amounts as a tool in taxonomy of Eucoccida is discussed.

Apicomplexa↗
PubMed1980

The development of Theileria=Cytauxzoon taurotragi (Martin and Brocklesby, 1960) from eland in its tick vector Rhipicephalus appendiculatus.

The sexual cycle of Theileria taurotragi was identified in the gut lumen of replete Rhipicephalus appendiculatur nymphs maintained at 28 degrees C which had fed on eland with rising parasitaemias. Macro- and micro-gametes developed from ring-form piroplasms within 24 h after repletion. The microgamonts were elongate with 2 or more lateral projections. The nuclei of the microgamonts divided into 4 and the microgamonts differentiated into 4 thread-like microgametes each with a central nucleus. Round macrogametes developed at the same time and stages indicative of the fusion of the macro- and microgametes were observed after 48 h. The resultant zygotes were detected in the gut wall cells by day 4, many being rounded and vacuolated with a peripheral nucleus. By day 7 the zygote cytoplasm became dense and they lay in clusters still within the cells. Binucleate zygotes were observed at this stage. The zygotes increased in size and by day 12 began to transform into kinetes by invagination. By the time the nymphs moulted into adults (about day 14), the kinete straightened to a broad anterior end and a tapering posterior with a mean length of 22.1 micrometer. Kinetes were detected in the haemolymph by day 16. By day 20 the kinetes had penetrated the salivary gland acinar cells where they underwent schizogony until the infected acinar cells were filled with multinucleated sporoblasts. The nuclei of the infected acinar cells became greatly enlarged soon after penetration of the kinete. Division of the sporoblast nuclei was stimulated by feeding of the adult ticks. From the 2nd day of attachment of the ticks to rabbits the sporoblasts underwent a process of schizogony to produce cytomeres. Each nucleus of the cytomere divided to produce several small nuclei which differentiated into uninucleated sporozoites.

Animals↗
PubMed1987

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.

Animals↗
PubMed1987

Experimental induction of Theileria parva lawrencei carrier state in an African buffalo (Syncerus caffer).

An African buffalo (Syncerus caffer), born in captivity and demonstrated to be Theileria-free, and 2 susceptible cattle were inoculated with a Theileria parva lawrencei sporozoite stabilate. The buffalo had a very mild disease reaction, while the 2 cattle died of acute theileriosis. It was possible to isolate T. p. lawrencei from the buffalo up to 888 days after infection by the application of non-infected Rhipicephalus appendiculatus nymphs and up to 657 days after infection by the establishment of lymphoblastoid cell lines infected with T. p. lawrencei schizonts from peripheral mononuclear blood cells. The infection rate and levels of Theileria in the resultant adult ticks varied from 11 to 70% with 0.3-11 acini infected/tick. Stabilates prepared from these tick batches caused fatal T. p. lawrencei infections in cattle.

Animals↗
PubMed1987

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)

Animals↗
PubMed1985

Intraerythrocytic schizogony of Theileria annulata.

The intraerythrocytic multiplication of two strains of Theileria annulata was studied with parasites maintained in stationary cultures and in the blood of infected cattle. In cultures established with blood from infected cattle 20-60% of single T. annulata piroplasms divided into quadruplet forms by day 6 in vitro. Transmission electron microscopic studies of T. annulata in culture showed that piroplasms possess intracytoplasmic food vacuoles and cytostomes during a pre-division trophozoite stage. The onset of intraerythrocytic multiplication was marked by the appearance of rhoptries and electron-dense plaques beneath the parasite's plasmalemmal membrane. The plaques developed into short segments of subplasmalemmal double membranes which were closely associated with the rhoptries. It was concluded that multiplication of T. annulata in erythrocytes occurred by schizogony, as nuclear division preceded cytoplasmic division and the final separation of merozoites. The four merozoites produced by intraerythrocytic schizogony had the same ultrastructural features as the T. annulata merozoites produced by intralymphocytic schizogony. Clusters of four merozoites, identical to those observed in stationary cultures, were also seen in the erythrocytes of persistently infected cattle and appeared to represent the most significant dividing forms of T. annulata in vivo.

Animals↗
PubMed1988

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.

Animals↗
PubMed1988

The effect of temperature on the rate of transmission of Theileria parva parva infection to cattle by its tick vector, Rhipicephalus appendiculatus.

A steer was infected with Theileria parva parva Kilae stabilate; nymphal Rhipicephalus appendiculatus ticks were applied to its ears so that they completed repletion when the steer had a high piroplasm parasitaemia. The engorged nymphs were subsequently incubated at 28 degrees C for 26-29 days to complete moulting, when the adult ticks were divided into two groups; one was incubated at 18 degrees C for 20 days and the other at 18 degrees C for 14 days and then at 37 degrees C for 6 days. Groups of ticks incubated at 37 and 18 degrees C were triturated and each resultant supernatant fluid inoculated into a steer. Both steers became infected, but the 37 degrees C supernatant group showed a much shorter pre-patent period to schizonts. Groups of ticks incubated at 37 or 18 degrees C were applied to pairs of cattle for 24, 48 and 72 h and then removed. There was a more rapid transmission of theileriosis to cattle by ticks kept at high ambient temperatures compared to those kept at low ambient temperatures. All cattle on which ticks treated at 37 degrees C were applied developed acute and fatal T. parva infection irrespective of the duration of tick application, while only 1 animal receiving ticks treated at 18 degrees C and fed for 72 h developed infection. The pre-patent period for macroschizonts was very short in all the groups receiving ticks incubated at 37 degrees C.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗