Abstract
Encystment is crucial for defense and reproduction in Cryptocaryon irritans. Therefore, understanding the encystment-related events in the protomont stage can help prevent and control C. irritans. Autophagy promotes protozoan parasite encystation. However, 3MA can inhibit autophagy. In this study, the effects of autophagy inhibition on encystation, survival rate, ultrastructural features, and metabolomic profiles of C. irritans, were evaluated using protomonts treated with 3MA (20 mM). The treatment with 3MA for about 4 h significantly lowered survival and encystation rates of protomonts to about 86.44% and 76.08%, respectively. Microstructural observations showed that the 3MA-treated protomonts showed deformed cell membranes and the cytoplasmic content spill. Furthermore, observation of the ultrastructure of 3MA-treated protomonts showed the destruction of organelles (Golgi bodies and mucocyst) and a lack of autophagosomes. However, no abnormality was observed in the control experiments. Furthermore, the metabolic analysis revealed suppression of metabolites, such as lipids, amino acids, and carbohydrates. These results demonstrate that 3MA can inhibit autophagy in C. irritans, thus hindering encystation, suppressing the metabolism of metabolites, and altering morphological ultrastructure in these parasites.
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Acknowledgements
The completion of this manuscript could not have been possible without the support and guidance of my all lab teachers and colleagues Liyao Zhou, Ruiling Zhou, and Lingling Wang. They had helped me for purchasing experimental tools. Their contributions are sincerely appreciated and gratefully acknowledged. Also, we deeply acknowledge funding support.
Funding
This work was supported by the Key program of Science and Technology of Zhejiang Province (grant No.2021C02024), the Ningbo Public Welfare Project (grant No.2022S159; 202002N3042),the Shanghai Natural Science Foundation (grant No. 17ZR1439600), the Shanghai Natural Science Foundation (grant No. 17ZR1439600), and the National Key Research and Development Project of China (grant No.2019YFD0900102).
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Bushra, Ivon F. Maha, Xiao Xie, and Fei Yin conceived and designed the study. Ivon F. Maha, Bushra, Youbin Yu, and Linhua Jiang performed animal feeding and laboratory experiments, analyzed data, interpreted the data, and prepared figures. Bushra and Ivon F. Maha wrote the manuscript and contributed equally to the study. Fei Yin, Summing Zhou, and Xiao Xie interpret the data and revise the manuscript. All authors read and approved the final manuscript.
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All animal and parasite experiments were performed according to local and central government regulations. The Institutional Animal Care and Use Committee of the Ningbo University approved all experiments. Informed consent was obtained from all individual participants included in the study.
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Bushra, Maha, I.F., Yu, Y. et al. Effects of autophagy inhibition by 3-methyladenine on encystation, morphology, and metabolites of Cryptocaryon irritans. Parasitol Res 122, 509–517 (2023). https://doi.org/10.1007/s00436-022-07751-w
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DOI: https://doi.org/10.1007/s00436-022-07751-w