Molecular characterization and analysis of drug resistance-associated protein enolase 2 of Eimeria tenella
Eimeria tenella, an intestinal parasite, has brought huge economic losses to the poultry industry. The prevalence and severity of the development of drug resistance has increased the challenge of coccidiosis control. We previously identified the enolase 2 of E. tenella (EtENO2) was differentially ex...
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Elsevier
2023-04-01
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Series: | International Journal for Parasitology: Drugs and Drug Resistance |
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Online Access: | http://www.sciencedirect.com/science/article/pii/S2211320723000040 |
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author | Yu Yu Wenhao Huang Qingjie Wang Hui Dong Qiping Zhao Shunhai Zhu Bing Huang Hongyu Han |
author_facet | Yu Yu Wenhao Huang Qingjie Wang Hui Dong Qiping Zhao Shunhai Zhu Bing Huang Hongyu Han |
author_sort | Yu Yu |
collection | DOAJ |
description | Eimeria tenella, an intestinal parasite, has brought huge economic losses to the poultry industry. The prevalence and severity of the development of drug resistance has increased the challenge of coccidiosis control. We previously identified the enolase 2 of E. tenella (EtENO2) was differentially expressed in drug-sensitive (DS) and drug-resistant strains using RNA-seq. In this study, the expression of EtENO2 in diclazuril-resistant (DZR), maduramicin-resistant (MRR), and salinomycin-resistant (SMR) strains was analyzed by quantitative real-time PCR (qRT-PCR) and western blots. EtENO2 was highly expressed in several drug-resistant strains compared with the DS strain. The qRT-PCR showed that the transcription level of EtENO2 in the field-isolated resistant strains was upregulated compared with the DS strain. The enzyme activity results indicated that the catalytic activity of EtENO2 in the drug-resistant strains was higher than in the DS strain. In addition, qRT-PCR and western blots showed that the expression level of EtENO2 was higher in second generation merozoites (SM) and unsporulated oocysts (UO) than that in sporozoites (SZ) and sporulated oocysts (SO). Immunofluorescence localization revealed that EtENO2 was distributed throughout SZ and SM and on the surface of the parasites. After the SZ invasion DF-1 cells, it was also observed on the parasitophorous vacuole membrane. Our secretion experiments found that EtENO2 could be secreted outside the SZ. This study indicated that EtENO2 might be related to the interaction between E. tenella and host cells and be involved in the development of E. tenella resistance to some anticoccidial drugs. |
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series | International Journal for Parasitology: Drugs and Drug Resistance |
spelling | doaj.art-7dbec1feb8c145b5bfc91b98cf4c01202023-03-31T05:53:20ZengElsevierInternational Journal for Parasitology: Drugs and Drug Resistance2211-32072023-04-01218190Molecular characterization and analysis of drug resistance-associated protein enolase 2 of Eimeria tenellaYu Yu0Wenhao Huang1Qingjie Wang2Hui Dong3Qiping Zhao4Shunhai Zhu5Bing Huang6Hongyu Han7Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal Parasitology of Ministry of Agriculture, Minhang, Shanghai, 200241, PR ChinaShanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal Parasitology of Ministry of Agriculture, Minhang, Shanghai, 200241, PR ChinaShanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal Parasitology of Ministry of Agriculture, Minhang, Shanghai, 200241, PR ChinaShanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal Parasitology of Ministry of Agriculture, Minhang, Shanghai, 200241, PR ChinaShanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal Parasitology of Ministry of Agriculture, Minhang, Shanghai, 200241, PR ChinaShanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal Parasitology of Ministry of Agriculture, Minhang, Shanghai, 200241, PR ChinaShanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal Parasitology of Ministry of Agriculture, Minhang, Shanghai, 200241, PR ChinaCorresponding author.; Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal Parasitology of Ministry of Agriculture, Minhang, Shanghai, 200241, PR ChinaEimeria tenella, an intestinal parasite, has brought huge economic losses to the poultry industry. The prevalence and severity of the development of drug resistance has increased the challenge of coccidiosis control. We previously identified the enolase 2 of E. tenella (EtENO2) was differentially expressed in drug-sensitive (DS) and drug-resistant strains using RNA-seq. In this study, the expression of EtENO2 in diclazuril-resistant (DZR), maduramicin-resistant (MRR), and salinomycin-resistant (SMR) strains was analyzed by quantitative real-time PCR (qRT-PCR) and western blots. EtENO2 was highly expressed in several drug-resistant strains compared with the DS strain. The qRT-PCR showed that the transcription level of EtENO2 in the field-isolated resistant strains was upregulated compared with the DS strain. The enzyme activity results indicated that the catalytic activity of EtENO2 in the drug-resistant strains was higher than in the DS strain. In addition, qRT-PCR and western blots showed that the expression level of EtENO2 was higher in second generation merozoites (SM) and unsporulated oocysts (UO) than that in sporozoites (SZ) and sporulated oocysts (SO). Immunofluorescence localization revealed that EtENO2 was distributed throughout SZ and SM and on the surface of the parasites. After the SZ invasion DF-1 cells, it was also observed on the parasitophorous vacuole membrane. Our secretion experiments found that EtENO2 could be secreted outside the SZ. This study indicated that EtENO2 might be related to the interaction between E. tenella and host cells and be involved in the development of E. tenella resistance to some anticoccidial drugs.http://www.sciencedirect.com/science/article/pii/S2211320723000040CoccidiaEimeria tenellaEnolase 2DiclazurilMaduramicinResistant strains |
spellingShingle | Yu Yu Wenhao Huang Qingjie Wang Hui Dong Qiping Zhao Shunhai Zhu Bing Huang Hongyu Han Molecular characterization and analysis of drug resistance-associated protein enolase 2 of Eimeria tenella International Journal for Parasitology: Drugs and Drug Resistance Coccidia Eimeria tenella Enolase 2 Diclazuril Maduramicin Resistant strains |
title | Molecular characterization and analysis of drug resistance-associated protein enolase 2 of Eimeria tenella |
title_full | Molecular characterization and analysis of drug resistance-associated protein enolase 2 of Eimeria tenella |
title_fullStr | Molecular characterization and analysis of drug resistance-associated protein enolase 2 of Eimeria tenella |
title_full_unstemmed | Molecular characterization and analysis of drug resistance-associated protein enolase 2 of Eimeria tenella |
title_short | Molecular characterization and analysis of drug resistance-associated protein enolase 2 of Eimeria tenella |
title_sort | molecular characterization and analysis of drug resistance associated protein enolase 2 of eimeria tenella |
topic | Coccidia Eimeria tenella Enolase 2 Diclazuril Maduramicin Resistant strains |
url | http://www.sciencedirect.com/science/article/pii/S2211320723000040 |
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