Transgenic
Eimeriamitis Expressing Chicken IL-4 Mediated Decrease
in Pathogenicity Compared to Wild Type Eimeria mitis Strains
in Broiler Chickens
Jamal Muhammad Khan1,2, Saeed
El-Ashram, Hong Bin Liu3,
Sher Hayat Khan4, Adnan Ayan5,
Xian Yong Liu1,2, Hui Wang2, Xin Ming Tang1,2,
Xun Suo1,2* and
Mohammad Farooque Hassan4,6
1State
Key Laboratory of Agro-Biotechnology; 2National Animal
Protozoa Laboratory & College of Veterinary Medicine, China
Agricultural University, Beijing, 100193, China; 3Department
of Pharmacology, Hebei North University, Zhangjiakou, Hebei, 075000,
China; 4Key laboratory of Animal Epidemiology and
Zoonosis; 4National Animal TSE Lab, College of Veterinary
Medicine, China Agricultural University, Beijing, 100193, China;
5Department of Parasitology, Faculty of Medicine, Adnan
Menderes University, Aydin, Turkey; 6Shaheed Benazir Bhutto
University of Veterinary and Animal Sciences, Sakrand, Sindh,
Pakistan *Corresponding author:suoxun256@163.com
Abstract
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.
Key words:
Birds,
Hematology,
Interleukin 4, Transgenic E. mitis