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The life cycle of Ascogregarina chagasi in larvae and in adult female Lutzomyia longipalpis was studied by light and electron microscopy. Sporozoites and young gamonts were attached to the epithelial lining of the larval midgut via an osmiophilic contact zone. The mucron of young gamonts was bordered by an invaginated pellicular fold, and an electron-opaque vesicular structure was observed adjacent to it. Sporozoites possessed an apical complex and were bound by a double membrane with underlying subpellicular microtubules. The gamont pellicle was uniformly corrugated and consisted of two cortical membranes and a plasma membrane. Mature gamonts and gametocysts were found in the posterior ectoperitrophic space of third and fourth instar larval midguts and in the haemocoel of adult flies. Gametocysts in adult females adhered to the genital accessory glands, where they were encased in electron-dense capsules secreted by the fly through haemocyte-mediated humoral immune reactions. Oocytes were spindle-shaped and bound by a double-layered wall with a discernible polar plug at each end. Sporulation was endogenous, occurring within gametocysts in the midguts of larvae or the accessory glands of adult females. FITC-phalloidine staining of all stages of A. chagasi except mature gametocysts produced bright fluorescence, indicating the presence of a diffuse, actin-like protein in the cytoplasm.
The sporozoans comprise a coherent group of protozoans, with characteristic and complex life cycles, containing 4-5000 species parasitic in invertebrates, particularly annelids and arthropods, and vertebrates. The group is a very successful one but neither its origins nor evolution are well understood. Considerations of traditional life cycles combined with newer methodologies have thrown some light on the evolutionary expansions of the main groups of sporozoans, the gregarines, coccidia, haemosporidians and piroplasms. The sporozoans of economic importance such as the coccidia, malaria parasites and piroplasms have received most attention but the data obtained have also thrown new light on the possible evolution of less well studied groups and it is concluded that conclusions based on simple comparisons of life cycles will have to be modified. It is also clear that humans have played a major part in affecting the distribution and present abundance of many sporozoans of economic significance and probably also those of less importance, and that the rates of evolutionary expansion are much more rapid than previously thought.
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The weight changes, carrier state and nature of chronic disease in cattle following infection with five isolates of Theileria parva and treatment with parvaquone were studied over an 18 month period while the cattle were maintained on normal management under strict tick control. Cattle infected with the T.p. parva Pugu II isolate gained weight similarly to uninfected control cattle and did not develop a detectable carrier state or show chronic disease. The cattle in the 4 other isolate groups regained the weight lost during the initial infection more slowly, then gained weight in parallel to the control cattle. Transmission of infection through ticks was achieved consistently from the T.p. parva Mbita I and Entebbe II isolate infections and intermittently from the T.p. lawrencei Mara II and Manyara infections. Infections were transmitted from cattle which had neither a detectable parasitaemia nor an antitheilerial antibody titre. Persistent macroschizont parasitoses were detected with the T.p. parva Entebbe II and the two T.p. lawrencei infections. The histopathology of the chronic disease lesions is described and the importance of the carrier state discussed.
Clinical, gross post mortem, histological and electron-microscopical data were collected from cats serially killed following experimental infection with Cytauxzoon felis. The agent was originally isolated from domestic cats with naturally occurring fatal cytauxzoonosis. Morphologically, the blood phase of feline Cytauxzoon appeared similar or identical to that described for cases of cytauxzoonosis in African ruminants and closely resembled also the blood phases of Theileria spp. and Babesia spp. The tissue phase, which occurred as schizont and merozoite stages within mononuclear phagocytes, was also similar to that seen in African cytauxzoonosis and in theileriosis. The tissue host cells, mononuclear phagocytes, appeared identical in feline and African cases. The Cytauxzoon host cells, however, were distinct from the tissue host cell of Theileria, in which schizogeny takes place within lymphocytes. Another difference was the lack of evidence of Cytauxzoon host-cell division which has been described for Theileria parva.
Protozoal encephalitic lesions were found in four aborted fetuses and one dead newborn calf. The organism was identified as Neospora caninum by immunoperoxidase. The brain lesions were of two forms. One was observed in three fetuses of 5 months gestation and was characterized by multifocal necrosis. The other was found in a 7-month fetus and in a newborn calf, and showed severe infiltration with macrophages and plasma cells containing IgG. This association, between the age of fetus and inflammation, may reflect development of the immune system in bovine fetuses.
We describe detailed methods for derivation and cloning of myeloma hybrids which secrete antibodies specific for antigens of protozoan parasites. The methods were designed to enable the derivation of large numbers of specific monoclonal antibodies and to give high cloning efficiencies of desired hybrids. Although special attention is paid to derivation and detection of anti-parasite antibodies, the methods can be applied to many different antibody-antigen systems. Using the described methods we have isolated more than 90 myeloma hybrids which secrete antibodies specific for antigens of African trypanosomes and Theileria parasites, thus illustrating their effectiveness.
A hybridoma cell line has been previously produced which secretes monoclonal antibodies able to neutralize sporozoites of Theileria parva, the causative agent of East Coast fever of cattle. Cells from this line were injected intra-peritoneally into pristane-treated BALB/c mice. During the last 4 days of hybridoma cell growth, mice were given 4 daily intraperitoneal injections of a mixture of tritiated amino acids in order to biosynthetically radiolabel the monoclonal antibody being produced in ascites fluid. The specific activity of the antibody obtained was 100 mCi/mmol. The labelled antibody was used to detect, by autoradiography, a surface coat antigen of T. parva sporozoites in cryostat sections of Theileria-infected tick salivary glands. The method allows the preparation of large quantities of biosynthetically radiolabelled immunological probes for the detection of immunoreactive sites in biological specimens.
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