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PubMed1978

In vitro studies on the action of menoctone and other compounds on Theileria parva and T. annulata.

Bovine lymphoblastoid cell cultures permanently infected with Theileria parva and T. annulata were used in an in vitro screen to test a wide range of compounds for chemotherapeutic activity against these parasites. Only two compounds, menoctone and methotrexate, showed significant activity, which has subsequently been confirmed by tests in infected cattle. Menoctone was about 100 times more active than methotrexate. Mixtures of menoctone and various other other compounds were tested, but showed no potentiation. The activity of menoctone was antagonized by cycloguanil and methotrexate. The in vitro test represents a rapid, cheap, and apparently reliable method of screening compounds for activity against Theileria.

Animals↗
PubMed1983

Isoelectric focusing in agarose: a highly discriminatory method for the detection of enzyme heterogeneity.

A comparison was made of the effectiveness of starch gel electrophoresis and isoelectric focusing in agarose for the separation of isoenzymes. Lactate dehydrogenase (LDH) of Theileria parva and glucose phosphate isomerase (GPI) of various cestodes were examined by the two techniques. Isoelectric focusing proved to be highly discriminatory, clearly distinguishing cestode orders and Taenia spp. which were closely similar by starch gel electrophoresis, and separating seven bands of Theileria-specific LDH activity which were not separable from normal bovine control bands on starch gels. The routine use of isoelectric focusing in agarose is therefore recommended for isoenzyme analysis.

Animals↗
PubMed1984

Evaluation of a range of antimicrobial agents against the parasitic protozoa, Plasmodium falciparum, Babesia rodhaini and Theileria parva in vitro.

Eighteen antimicrobials commonly used in tissue culture were screened in three different protozoan test systems in order to establish their suitability for routine inclusion in protozoal cultivation systems. The human malaria parasite, Plasmodium falciparum, was inhibited by more than half the antibiotics tested at concentrations recommended for normal tissue culture use. Eight compounds were well tolerated and thus could be used prophylactically to prevent microbial contamination. These antimicrobials were the bactericidal aminoglycoside antibiotics, streptomycin, gentamicin and kanamycin, the bacteriostatic protein synthesis inhibitors, chloramphenicol and chlortetracycline and the antifungals, 5-fluorocytosine, nystatin and amphotericin B. Babesia rodhaini and Theileria parva were less sensitive than P. falciparum and tolerated all 18 compounds at concentrations well above 100 micrograms ml-1. Extension of the study to examine direct antiprotozoal action of these and other antimicrobials not normally used in culture confirmed that P. falciparum was significantly more sensitive than the other parasites. Tylosin, rifamycin, gramicidin D and valinomycin were all strongly antimalarial with IC50 values of 0.245, 1.20, 1.3 X 10(-3) and 1.9 X 10(-3) micrograms ml-1 respectively. This compares with a value of 1.35 X 10(-2) micrograms ml-1 for the standard antimalarial, chloroquine. Only valinomycin and, more particularly, gramicidin D were significantly active against B. rodhaini and T. parva. Gramicidin D was more effective, but more toxic, than the standard antiprotozoal agents tested at curing in vivo malarial and babesial infections in mice.

Animals↗
PubMed1984

The development of Theileria annulata in the salivary glands of the vector tick Hyalomma anatolicum anatolicum.

The development at 28 degrees C of Theileria annulata (Hissar) in the salivary glands of its tick vector, Hyalomma anatolicum anatolicum, was studied using Giemsa-stained smears, methyl green-pyronin-stained preparations of whole glands, and electron microscopy. Nymphs which had engorged on T. annulata-infected calves showed kinetes in the haemolymph from Day 7 to Day 14 post engorgement, when all ticks had completed moult. Intracellular sporonts were observed within salivary gland acini from Day 7 onwards and these developed by rapid nuclear division and cytoplasmic proliferation to form primary sporoblasts. Further development was stimulated by feeding on a rabbit or by incubation at 36 degrees C. The primary sporoblasts appeared to become organized into membrane-bound subunits. Within 48 hours of attachment to the host, or 72 hours of 36 degrees C incubation, these units dissociated to form secondary sporoblasts. The final phase of development resulted in the progressive formation of discrete, uninuclear sporozoites within these secondary sporoblasts. No morphological differences were observed in parasite maturation between the fed and incubated groups although development was retarded by at least 24 hours in the latter. In the incubated group there was also a marked decrease in the degree of synchrony of development which resulted in fewer sporozoites being present at any one time.

Animals↗
PubMed1985

The maintenance and survival of Theileria annulata in colonies of Hyalomma anatolicum anatolicum.

Procedures are described for the colonization of Hyalomma anatolicum anatolicum ticks using gerbils, rabbits and cattle as hosts. On rabbits H. a. anatolicum undergoes a two-host cycle but methods are described for obtaining either unfed or engorged nymphs. Data are given on the life cycle timings and the numbers and timings of tick feeding regimes. The infection of H. a. anatolicum with two strains of Theileria annulata is described and methods given for monitoring the development and survival of T. annulata in H. a. anatolicum. Data are presented indicating the optimum maintenance conditions for T. annulata in H. a. anatolicum. Piroplasm parasitaemias in cattle greater than 2% gave high infection rates in adult ticks if engorged nymphs were moulted at 28 degrees C for 28 days. The Theileria will survive for ten months in such adults stored at 12 degrees C after moult, and when fed the ticks will produce maximum numbers of sporozoites on the third day of feeding, whatever their age. The moult of engorged nymphs is retarded at 18 degrees C but the Theileria in such ticks will develop normally when the ticks are moulted at 28 degrees C after four months storage at 18 degrees C.

Animals↗
PubMed1985

The presence of Theileria sergenti in Haemaphysalis longicornis overwintering in pasture in Japan.

Unfed Haemaphysalis longicornis ticks were collected from the soil of pasture during the winter and their salivary glands were examined by the methyl green pyronin staining method in order to detect overwintering of Theileria sergenti, the causative agent of bovine theileriosis in Japan. A transmission experiment was carried out using these ticks applied to a splenectomized calf. The protozoa which appeared in the peripheral blood were identified parasitologically and seroimmunologically as T. sergenti. The results suggested that T. sergenti in Japan might overwinter in H. longicornis which moulted after repletion on infected cattle in the autumn and hid in the soil during the winter. Moreover, it seemed possible that T. sergenti might develop to the infective stage in H. longicornis during the overwintering period.

Animals↗