Transgenic Eimeria mitis expressing chicken IL-4 mediated decrease in pathogenicity compared to wild type Eimeria mitis strains in broiler chickens.

Jamal Muhammad Khan; Saeed El-Ashram; Hong Bin Liu; Sher Hayat Khan; Adnan Ayan; Xian Yong Liu; Hui Wang; Xin Ming Tang; Xun Suo; Mohammad Farooque Hassan

Jamal Muhammad Khan State Key Laboratory of Agro-Biotechnology, China Agricultural University, Beijing, 100193, China;National Animal Protozoa Laboratory & College of Veterinary Medicine, China Agricultural University, Beijing, China

Saeed El-Ashram Department of Pharmacology, Hebei North University, Zhangjiakou, Hebei, China

Hong Bin Liu Department of Pharmacology, Hebei North University, Zhangjiakou, Hebei, China

Sher Hayat Khan Key laboratory of Animal Epidemiology and Zoonosis, College of Veterinary Medicine, China Agricultural University, Beijing, China; National Animal TSE Lab, College of Veterinary Medicine, China Agricultural University, Beijing, 100193, China

Adnan Ayan Department of Parasitology, Faculty of Medicine, Adnan Menderes University, Aydin, Turkey

Xian Yong Liu State Key Laboratory of Agro-Biotechnology, China Agricultural University, Beijing, 100193, China; National Animal Protozoa Laboratory & College of Veterinary Medicine, China Agricultural University, Beijing, China

Hui Wang National Animal Protozoa Laboratory and College of Veterinary Medicine, China Agricultural University, Beijing, China

Xin Ming Tang State Key Laboratory of Agro-Biotechnology, China Agricultural University, Beijing, China; National Animal Protozoa Laboratory & College of Veterinary Medicine, China Agricultural University, Beijing, China

Xun Suo State Key Laboratory of Agro-Biotechnology, China Agricultural University, Beijing, China; National Animal Protozoa Laboratory & College of Veterinary Medicine, China Agricultural University, Beijing, China

Mohammad Farooque Hassan Key laboratory of Animal Epidemiology and Zoonosis, College of Veterinary Medicine, China; National Animal TSE Lab, College of Veterinary Medicine, China Agricultural University, Beijing, 100193, China; Shaheed Benazir Bhutto University of Veterinary and Animal Sciences, Sakrand, Sindh, Pakistan

The present study was designed to establish transgenic Eimeria mitis (tE. mitis) expressing chicken interleukin-4 (ChIL-4) and further exploitation of tE. mitis as a novel vaccine vector to control coccidiosis. For this purpose we compared the clinico-hematological alterations induced by wild type and tE. mitis carrying ChIL- 4 in broilers birds. Wild type E. mitis (wE. mitis) oocysts were nucleofected with double cassette vector and transgenic oocysts were propagated for stable transfection. Stability of the transfected E. mitis was observed by fluorescent microscopy and PCR analysis revealed the appearance of 411bp band suggestive of presence of IL-4 into the genome of tE. mitis. Birds inoculated with transgenic EmiChIL-4 excreted fewer oocysts and fecundity potential of tE. mitis turned out to be significantly lower than that of the wild type parent strain. Moderate clinical and behavioral signs such as restlessness, diarrhea, emaciation, decreased feed intake and loss of body weight was observed in birds inoculated with wE. mitis oocysts. The erythrocyte counts, Hb conc, PCV and MCHC decreased in birds infected with wE. mitis as compared to those infected with tE. mitis. The total leukocyte count including heterophils, eosinophil and basophils increased while lymphocytes were decreased in birds inoculated with wE. mitis as compared to birds in control group and those infected with tE.mitis. It was concluded that tE. mitis incorporated with ChIL-4 functions well as an adjuvant, conferring reduced fertility and compromised pathogenicity of tE. mitis. This discrimination, may be applied as a potential vaccine vector to control coccidiosis as well as pathogens from which antigens are expressed by tE. mitis.