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Updated: Feb 6, 2026
Nucleofection and In Vivo Propagation of Chicken Eimeria Parasites
Published on: February 14, 2020
The molecular characterization and protective efficacy of microneme 3 of Eimeria mitis in chickens
Xinmei Huang1, Jianhua Liu2, Di Tian2
1MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, PR China; Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Science, Nanjing 210014, PR China.
Abstract:
E. mitis is ubiquitous in clinical coccidiosis caused by mixed infection of Eimeria species and the infection by E. mitis usually significantly impairs productivity of the infected chickens. To date, however, few protective antigens from E. mitis have been reported. In this study, the molecular characterization and protective efficacy of microneme 3 of Eimeria mitis (EmiMIC3) were analyzed. EmiMIC3 gene was cloned from sporozoites of E. mitis and its MARs (microneme adhesive repeats domain) were predicted. Recombinant EmiMIC3 (rEmiMIC3) was expressed in E. coli and purified and then was analyzed by western blot with anti-E. mitis chicken serum. Meanwhile, native EmiMIC3 from sporozoites was analyzed by anti-rEmiMIC3 rat serum. The expressions of EmiMIC3 in E. mitis sporozoites and merozoites were analyzed by immunofluorescence assay. The rEmiMIC3-induced changes of T lymphocytes subpopulation, serum cytokines and IgY levels and the protective efficacy of rEmiMIC3 were determined in animal experiments. The results showed that the deduced open reading frame (ORF) of EmiMIC3 was composed of 1145 amino acids, possessing 9 MARs. EmiMIC3 gene was submitted to GenBank (accession number: MG888670). EmiMIC3 could express in sporozoites and merozoites respectively and located at the apex of E. mitis sporozoite. Western blot assay revealed that the rEmiMIC3 could be recognized by serum of chicken infected by E. mitis and the native EmiMIC3 from sporozoites could also be recognized by rat serum against rEmiMIC3. Following vaccination with rEmiMIC3, higher levels of IL-10, IFN-γ, TGF-βand IL-17, higher proportions of CD4+/CD3+ and CD8+/CD3 + T lymphocytes and higher level of IgY antibody were induced compared to the controls. Vaccination with rEmiMIC3 prominently increased the weight gains and decreased oocyst output of the vaccinated chickens after challenge infection. Our result not only enriches protective candidate antigen of E. mitis, but also provides available protective antigen of E. mitis for the development of multivalent vaccines against infection caused by mixture of Eimeria species in clinical coccidiosis.
Insights
This study identifies microneme 3 of Eimeria mitis (EmiMIC3) as a promising antigen for chicken coccidiosis vaccines. Vaccination with EmiMIC3 enhanced immune responses and improved chicken growth and reduced parasite shedding.
Area of Science:
- Veterinary Parasitology
- Immunology
- Molecular Biology
Background:
- Eimeria mitis infection significantly impacts chicken productivity in mixed Eimeria infections.
- Few protective antigens from E. mitis have been identified to date.
- Microneme proteins are crucial for Eimeria invasion and are potential vaccine candidates.
Purpose of the Study:
- To characterize the Eimeria mitis microneme 3 (EmiMIC3) gene and protein.
- To evaluate the protective efficacy of recombinant EmiMIC3 (rEmiMIC3) as a vaccine against E. mitis infection in chickens.
Main Methods:
- EmiMIC3 gene was cloned and its microneme adhesive repeats (MARs) were predicted.
- Recombinant EmiMIC3 (rEmiMIC3) was expressed in E. coli and purified.
- Immunofluorescence, Western blot, and animal experiments were used to analyze EmiMIC3 expression, antigenicity, and vaccine efficacy.
Main Results:
- The EmiMIC3 gene (GenBank accession: MG888670) encodes a 1145-amino acid protein with 9 MARs, expressed in sporozoites and merozoites.
- rEmiMIC3 was recognized by chicken anti-E. mitis serum and native EmiMIC3 by anti-rEmiMIC3 rat serum.
- rEmiMIC3 vaccination induced higher levels of cytokines (IL-10, IFN-γ, TGF-β, IL-17), T lymphocytes (CD4+/CD3+, CD8+/CD3+), and IgY antibodies, leading to improved weight gain and reduced oocyst output post-challenge.
Conclusions:
- EmiMIC3 is a potential protective antigen for E. mitis.
- This study provides a valuable antigen for developing multivalent vaccines against mixed Eimeria infections in poultry.
- EmiMIC3 vaccination demonstrates significant protective effects against E. mitis challenge infections.
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