Search

Development of Eimeria tenella was studied in primary cultures of chicken kidney cells maintained in Medium 199 lacking each of the following: vitamin A, biotin, p-aminobenzoic acid, folic acid, nicotinamide, Ca pantothenate, pyridoxine, pyridoxal, riboflavin, thiamin, ascorbic acid, calciferol, alpha-tocopherol, and menadione. Data obtained concerning numbers of mature schizonts or total numbers of parasites or both indicated that all of the vitamins are needed for 1st- and 2nd-generation schizogony, and all except calciferol and folic acid are needed for gametogony.

Examination of littoral fish Blennius pholis and Cottus bubalis caught at Aberystwyth and Porth Cwyfan, Wales, U.K. revealed 2 species of coccidia. Eimeria dingleyi sp. n. Oocysts spherical (16.1-19.2) to subspherical (13.9-14.2 X 18.8-20.0) micron, with thin walls; sporulation outside the host to produce ellipsoid sporocysts; endogenous phases in epithelial cells throughout intestine; 26 of 58 B. pholis infected. Eimeria variabilis (Théohan) Reichenow. Oocysts spherical (11.9-14.6) to subspherical (9.2-10.9 X 13.9-14.3) micron, sporulation in lining of pyloric ceca and rectum; previously unrecorded schizonts and gametocytes present; 21 of 25 C. bubalis infected. Electron microscopy revealed that the oocyst wall of E. variabilis consists of a thin membrane whereas the sporocyst wall is thick and 3-layered. Typical oocyst wall-forming bodies were absent from the macrogamete. Cytochemical tests on the endogenous stages of E. dingleyi and E. variabilis indicated that in general they resembled other coccidia in their chemical constitution.

An assay has been developed using parasite-specific incorporation of 3H-uracil to assess the intracellular growth of Eimeria tenella in vitro. As shown by both scintillation counts and autoradiography, 3H-uracil was incorporated specifically into intracellular parasites from the onset of infection and continued throughout development of the first generation schizonts. Mature schizonts and first generation merozoites did not continue to incorporate additional 3H-uracil, indicating that RNA synthesis had halted in these stages. Based on these findings, a semi-automated microscale uracil incorporation assay was developed to determine parasite viability. This method should be useful for biochemical studies with intracellular parasites and for screening compounds for anticoccidial activity. The ease, rapidity, and quantitative nature of this assay contrasts favorably with standard morphometric approaches of determining parasite development. In addition, parallel studies using host cell incorporation of 3H-uridine have been introduced as a method of determining whether antiparasitic activity is direct or indirect in relation to effects on the host cell.