Abstract
Chicken coccidiosis is caused by the apicomplexan parasite Eimeria spp. At present, drug resistance of Eimeria is common because of the indiscriminate use of anticoccidial drugs. The gene encoding surface antigen 10 of Eimeria tenella (EtSAG10) is differentially expressed between drug-resistant and drug-sensitive strains. RNA-seq analysis indicated that this gene was downregulated in strains resistant to maduramicin and diclazuril compared to susceptible strains. EtSAG10 DNA sequence alignment revealed that they contained one and ten mutations in MRR and DZR, compared with DS, respectively. A full-length EtSAG10 cDNA was successfully cloned and expressed, and the polyclonal antibody was prepared. The transcription and translation levels of EtSAG10 were analyzed by quantitative real-time PCR (qPCR) and Western blotting. The localization of EtSAG10 in Spz, Mrz, and parasites in the first asexual stage was determined by indirect immunofluorescence. The potential association of EtSAG10 with sporozoite invasion of host cells was assessed by invasion inhibition assays. The results showed that EtSAG10 had a predicted transmembrane domain at the C-terminal end and a predicted signal peptide at the N-terminal end. EtSAG10 was downregulated in drug-resistant strains, which is consistent with the RNA-seq results. The EtSAG10 protein was localized to the parasite surface and parasitophorous vacuole membrane. This protein was shown to play a role in the infection of chicken intestine by sporozoites.
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We would like to thank all organizations which funded this work and all the teachers who cooperated in technical assistance.
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This work was supported by the National Key Research and Development Program of China (2018YFD0500302), the National Natural Science Foundation of China (Grant No. 31572266, No. 31672551), the National Sharing Service Platform for Parasite Resources (No. TDRC-22), and the Shanghai Minhang District talent development special funds.
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Fig. S1
Alignment of EtSAG10 DNA sequences of DS, DZR and MRR. (PNG 135 kb)
Fig. S2
Nucleotide sequence of the full-length cDNA and deduced amino acid sequence of EtSAG10. N-glycosylation site is shown in red. Casein kinase II phosphorylation site is indicated by the wavy underline. A putative N-myristoylation site is underlined. A protein kinase C phosphorylation site is shown in blue. The threonine-rich domain is in a rectangle. (PNG 198 kb)
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Liu, G., Zhu, S., Zhao, Q. et al. Molecular characterization of surface antigen 10 of Eimeria tenella. Parasitol Res 118, 2989–2999 (2019). https://doi.org/10.1007/s00436-019-06437-0
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DOI: https://doi.org/10.1007/s00436-019-06437-0