Research paperThe molecular characterization and protective efficacy of microneme 3 of Eimeria mitis in chickens
Section snippets
Introdction
Coccidiosis of domestic chickens, one of the major diseases in poultry farming, is caused by several species of the genus Eimeria and causes annual economic losses of more than $3 billion worldwide (Blake and Tomley, 2014; Witcombe and Smith, 2014). Infection of Eimeria parasite causes death and low productivity including inefficient feed utilization, poor weight gain and reduced egg production of the infected chickens. Subclinical infection of less-pathogenic Eimeria species, such as E. mitis
Animals and parasites
New-born Hy-Line layer chickens were raised under coccidia-free conditions for recovery and propagation of E. mitis parasite and experimental vaccination study. The chickens were provided with feed and water without anticoccidial drugs. SD rats of 30 days old were bought from the Comparative Medicine Centre, Yangzhou University, Yangzhou, China, for antiserum collection. All animal studies and protocols were approved by the Institutional Animal Care and Use Committee of Nanjing Agricultural
Cloning and sequence analysis of EmiMIC3
EmiMIC3 gene was amplified from the purified E. mitis sporozoite and submitted to GenBank with an accession number of MG888670. The open reading frame (ORF) is predicted to encode an 1145-amino acid protein with a molecular weight of 124.9 kDa and a pI (protein isoelectric point) of 4.71.
Characteristic analysis of MARs in EmiMIC3
BlastP revealed that EmiMIC3 contains 9 MARs, namely MAR1 (residues 49–142), MAR2 (residues 157–250), MAR3 (residues 315–408), MAR4 (residues 456 to 547), MAR5 (residues 569–662), MAR6 (residues 677–768), MAR7
Discussion
Avian coccidiosis is caused by individual or multiple Eimeria species and causes huge economic losses to poultry industry (Blake and Tomley, 2014; Witcombe and Smith, 2014). This disease is currently controlled through a combination of good husbandry, chemoprophylaxis and/or live parasite vaccination. However, next-generation vaccines including subunit or recombinant vaccines are required (Blake et al., 2017). To date, the reported candidate antigens for developing next-generation anticoccidial
Conclusion
E. mitis is one of the most prevalent Eimeria species causing mixed infection in backyard flocks and commercial flocks. MICs play important roles in parasite adhesion and invasion of host cells. To date, the reported protective microneme protein from E. mitis is not seen. In this study, an invasion related protein of EmiMIC3 was cloned and expressed. Its molecular characterization and protective efficacy were analyzed. EmiMIC3 possesses 9 microneme adhesive repeat regions. It is expressed in
Conflict of interest
The authors declare that they have no competing interests.
Acknowledgments
This work was supported by the Joint Research Project between National Natural Science Foundation of China and Pakistan Science Foundation (NSFC-PSF) (Grant No. 31661143017), the National Natural Science Foundation of China (Grant No. 31372428, 31672545) and the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD).
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