Pathological effects and reduced survival in Rhipicephalus appendiculatus ticks infected with Theileria parva protozoa.

Pathological effects have been demonstrated in a number of arthropod species as a consequence of parasitic infection. This is usually manifest as reduced arthropod survival and/or fecundity. This paper describes the detrimental effects the protozoan parasite, Theileria parva has on Rhipicephalus appendiculatus ticks. R. appendiculatus ticks were dissected and sectioned at regular intervals during their nymph to adult moult after detaching from a T. parva infected calf, and assessed by light and electron microscopy. The reproductive capacity of the T. parva infected ticks was also compared with that of control, uninfected ticks. The number of T. parva forms seen during the ticks' moult were recorded and showed a substantial reduction as the moult progressed. A non-linear relationship between piroplasm ingestion by the engorged nymph and eventual adult salivary gland infection levels was shown. Tick gut and salivary gland pathology was noted at various stages throughout the moult and correlated with the parasite burdens in the affected organs at those timepoints. The reproductive performance of infected ticks was greatly impaired compared to controls. Infected female ticks had longer bloodmeal engorgement times, reduced bloodmeal volumes, smaller egg batch weights and greatly decreased egg hatching success. The pathological effects are discussed in relation to parasite population dynamics within the ticks and compared with similar examples of pathology evidenced with other parasite infected arthropod species.

The use of nocodazole in cell cycle analysis and parasite purification from Theileria parva-infected B cells.

Theileria parasites transform bovine leukocytes and induce uncontrolled lymphoproliferation only in the macroschizont stage of their life cycle. The isolation of highly purified stage-specific parasite RNA and proteins is an essential prerequisite when studying the Theileria-host relationship. We therefore improved a protocol based on the cytolytic bacterial toxin aerolysin by taking advantage of the microtubule inhibitor nocodazole. In this report we describe that nocodazole-mediated separation of the parasite from the host cell microtubule network was used with success to improve quantity and quality of purified parasites. We furthermore show that nocodazole is a useful tool to study cell cycle checkpoints due to its capacity to induce reversible cell cycle arrest in Theileria-infected B cells.

The entry of Theileria parva merozoites into bovine erythrocytes occurs by a process similar to sporozoite invasion of lymphocytes.

The entry of Theileria parva merozoites into bovine erythrocytes in vivo is described and compared to sporozoite invasion of lymphocytes. Merozoites make initial contact with erythrocytes with any part of their surface and invasion of the host cell does not require the re-orientation of the apical end of the merozoite towards the surface of the erythrocyte. After the initial attachment the merozoite and host cell membranes form a continual close junction with the two apposed membranes separated by a 6-8 nm gap containing moderately dense material. The progressive circumferential 'zippering' of these closely apposed membranes leads to the movement of the parasite into the erythrocyte. The newly internalized merozoite which is completely surrounded by the erythrocyte plasma membrane escapes from this enclosing membrane by a process involving the discharge of at least the rhoptries; whether the merozoite also contain other types of secretory organelles (e.g. micronemes, microspheres or dense bodies) remains to be determined. Morphologically, the events involved in merozoite invasion of erythrocytes are almost identical to the process of sporozoite invasion of lymphocytes but differ significantly from the entry process of the invasive stages of other Apicomplexan parasites.

Differential development and emission of Theileria parva sporozoites from the salivary gland of Rhipicephalus appendiculatus.

The initiation of feeding of infected Rhipicephalus appendiculatus adults induces the rapid development of Theileria parva sporoblasts within the salivary gland acini leading to the production of numerous sporozoites which are inoculated into the mammalian host initiating infection. In this study the pattern of development, host cell specificity and emission of T. parva sporozoites within the salivary glands of heavily infected, 4-day fed adult R. appendiculatus ticks was examined. Infected acini were randomly distributed throughout the salivary gland. Sporozoite development within each gland was not synchronized and wide variation in the rate of parasite development, which correlated with the secretory activity of the individual acinus, was observed in all glands examined. Previous studies had shown that T. parva developed primarily in Type III 'e' cells. However, in heavily infected salivary glands sporogony and the emission of mature sporozoites also occurred in 'c' cells of Type II acini. Sporozoite emission from infected cells occurred by a process similar to apocrine secretion. The loss of the apical membrane of the infected cell allowed sporozoites free access to the lumen of the acinus and into the collecting ducts of the salivary gland. Sporozoite discharge was gradual since few parasites were found in the acinus valve or in the collecting ducts. Furthermore, the small size of the acinar valve aperature ensures that only small numbers of sporozoites can be released at any one time from an infected acinus.

How individual cells develop from a syncytium: merogony in Theileria parva (Apicomplexa).

The central problem for Theileria parva during merogony is how to form numerous individual, uninucleate merozoites from a syncytial schizont so that each merozoite contains a single nucleus and a prescribed assortment of organelles. The way T. parva packages all the requisite organelles into free merozoites is by binding these organelles to the nuclear envelope, which in turn becomes associated, both directly and through the rhoptry complex, with the schizont plasma membrane. Formation of the merozoites occurs in a synchronous manner by a budding process. The merozoites develop with the rhoptry complex at the apical end by the progressive, outward evagination of the schizont plasma membrane. This evagination of the plasma membrane is associated with, and presumably induced by, the development of an orderly array of tubules that originate from the apical end and progressively form a longitudinal basket enclosing first the rhoptry complex, then the mitochondria and ribosomes, and finally the nucleus. The process of merogony is compared to sporogony within the tick salivary gland and with the differentiation of the intra-erythrocytic piroplasm stage. Because all three processes occur by a morphologically similar mechanism, the possibility that the parasite uses a single cassette of genes to perform each of these similar processes is discussed.