ABSTRACT:The present paper reports on the IgA, IgG and IgM antibodies secreting cells (ASC) in the spleen of chickens vaccinated with E. tenella (local isolates) gametocyte vaccine(s) by using Enzyme Linked Immunospot (ELISPOT) assay. Irrespective of the vaccine used, the number of IgG antibody secreting cells (ASC) in the spleen of orally vaccinated chickens was higher than the number of IgA and IgM ASC. Maximum numbers of IgG, IgA and IgM ASC were found in chickens vaccinated with sonicated gametocyte formalin inactivated vaccine (Group III) followed by formalin inactivated gametocytes (Group II) and gametocytes (Group I). The number of ASC (IgG, IgA and IgM) per 10 4 cells in spleen was significantly higher (P < 0.05) in Group III as compared to Group II and Group I. Results of the challenge experiments revealed maximum protection against mixed species of the genus Eimeria in Group III (77.8%) followed by Group II (55.6%) and Group I (52.0%). Lesion scoring was directly proportional to the per cent mortality and oocysts per gram of droppings but inversely proportional to the per cent protection. Based on the results, it was assumed that the spleen in chickens is one of the major sources of cells producing IgA, IgG and IgM antibodies.
Keywords: Eimeria tenella; gametocytes; antibodies secreting cells; ELISPOTChickens infected with Eimeria species produce parasite-specific antibodies that are present in the systemic circulation and mucosal secretions (Lillehoj and Trout, 1996). Like in mammals, three principal classes of antibodies including IgM, IgA and IgG (IgY) are known in chickens (LebacqVerheyden et al., 1972). Upon exposure to Eimeria species, chickens produce all three classes of antibodies. The cells expressing E. tenella specific IgG have been detected in the lamina propria of the caeca and in the red pulp of the chicken spleen (Vervelde et al., 1992).The IgM secreting plasma cells in the lamina propria of the villi upon E. tenella infection have been detected (Jeurissen et al., 1989). An elevated level of IgA has been found in a tissue parasitized with E. tenella (Girard et al., 1997). The present paper reports on IgM, IgA and IgG antibody secreting cells in the spleen of chickens immunized with egg propagated E. tenella gametocyte vaccines.
MATERIAL AND METHODS
Ex-sporocystationSporulated oocysts of E. tenella (local isolates) maintained in Immunoparasitology Laboratory, University of Agriculture, Faisalabad, Pakistan were processed for excystation followed by exsporocystation to release sporozoites following the method of Speer et al. (1973). Briefly, purified sporulated oocysts were treated with 2.5% sodium hypochlorite for 20 min followed by continuous stirring in a vessel containing sterilized glass beads (425-600 µm diameter) for 25 min on a magnetic stirrer. The excysted material was centrifuged (310 × g for 10 min), the supernatant was discarded to remove sodium hypochlorite and the sediment was given three washings with phosphate-buffered saline (PBS). The washed sporocysts were suspended in...