Identification and characterization of a cDNA clone-encoding antigen of Eimeria acervulina

Eimeria spp. are the causative agents of coccidiosis, a major disease affecting the poultry industry. So far, only a few antigen genes of E. acervulina have been reported. In this study, a clone, named as cSZ-JN2, was identified from a cDNA expression library prepared from E. acervulina sporozoite stage with the ability to stimulate the chicken immune response. The sequence analysis showed that the open reading fragment (ORF) of cSZ-JN2 was 153 bp in size and encoded a predicted protein of 50 amino acids of Mr 5·3 kDa. BLASTN search revealed that cSZ-JN2 had no significant homology with the known genes of E. acervulina or any other organism (GenBank). The recombinant cSZ-JN2 antigen expressed in E. coli was recognized strongly by serum from chickens experimentally infected with E. acervulina. Immunofluorescence analysis using antibody against recombinant cSZ-JN2 indicated that this protein was expressed in sporozoite and merozoite developmental stages. Animal challenge experiments demonstrated that the recombinant protein of cSZ-JN2 and DNA vaccine carrying cSZ-JN2 could significantly increase the average body weight gains, decrease the mean lesion scores and the oocyst outputs of the immunized chickens and presented anti-coccidial indices of more than 165. All the above results suggested that the cSZ-JN2 was a novel E. acervulina antigen and could be an effective candidate for the development of a new vaccine against E. acervulina infection.