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Prevalence and Molecular Epidemiology of Cryptosporidium Infection in Clarias gariepinus Fish in Egypt

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Abstract

Purpose

This study investigated the prevalence and molecular detection of Cryptosporidium spp. in catfish (Clarias gariepinus).

Methods

A total of 300 Carias gariepinus fish were collected from two freshwater sources: the Nile River (180) and drainage canals (120). The stomach and intestine epithelium of each individual fish sample were screened by modified Ziehl–Neelsen (mZN) staining technique for the detection of Cryptosporidium oocysts followed by the serological survey for detection of Cryptosporidium antibodies using Enzyme-Linked Immunosorbent Assay (ELISA) and molecular characterization using complemented DNA polymerase chain reaction (cPCR).

Results

ELISA showed higher prevalence of 69.3% than that prevalence obtained by mZN, 64% for the total examined Clarias gariepinus fish. Also, higher prevalence of Cryptosporidium infection 65.5% and 75.8% obtained by ELISA than 61.1% and 68.3% by mZN, in both fish groups from Nile River and Drainage canal, respectively. PCR analysis revealed the expected positive bands at 1056 bp. DNA sequencing and phylogenetic analysis proved that the positive-PCR Cryptosporidium isolate identified in the present study was Cryptosporidium molnari.

Conclusion

Freshwater fishes (Clarias gariepinus) are subjected to a high infection rate with Cryptosporidium spp.; the drainage canals obtained fishes showed higher prevalence than that collected from Nile River which indicates an important public health problem and a potential risk of drainage canals in Egypt. ELISA showed higher prevalence of cryptosporidiosis than mZN, for the total examined Clarias gariepinus fish and phylogenetic analyses confirmed this protozoal organism to be a novel species of Cryptosporidium molnari.

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Acknowledgements

Authors are very grateful and extend their thanks and gratitude to the soul of the late Prof Dr. Mohey A. Hassanain, Professor of Parasitology, Department of Zoonotic Diseases, NRC, Egypt for his encouragement and valuable guidance to finish this work.

Funding

This research received no specific grant from any funding agency in the public, commercial, or not-for-profit sectors.

Author information

Author notes

  1. Fathy A. Abdel-Ghaffar

    Present address: Zoology Department, Faculty of Science, Cairo University, Giza, Egypt

Authors and Affiliations

  1. Department of Zoonotic Diseases, Veterinary Research Division, National Research Centre, El-Tahrir Street, Dokki, Post Box 12622, Giza, Egypt

    Raafat M. Shaapan

  2. Zoology Department, Faculty of Science, Cairo University, Giza, Egypt

    Kohar Garo Varjabedian & Gehad I. Saad-Alla

Authors
  1. Raafat M. Shaapan
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  2. Fathy A. Abdel-Ghaffar
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  3. Kohar Garo Varjabedian
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  4. Gehad I. Saad-Alla
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Contributions

Dr. RMS: planned and design the study, serological, molecular investigation, and drafting the paper. Dr. FAA: sharing in the idea and study design, identification of Cryptosporidium sp. and participated in drafting the manuscript. Dr. KGV: sharing in study conception, laboratory work, interpreted the data results, and helped in manuscript preparation and Dr. GIS: involved in samples collection and preparation, sharing serological and molecular tests. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Raafat M. Shaapan.

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Conflict of interest

The authors of the current work declare that they have no conflicts of interest in this study.

Ethical approval

The study was ethically cleared and approved by Medical Research Ethical Committee Research, National Statement on Ethical Conduct in Human and animals Research at National Research Centre, Egypt under registration number # 21-054#.

Significance

The freshwater fishes (Clarias gariepinus) are subjected to high infection rate with Cryptosporidium sp.; the drainage canals obtained fishes showed higher Cryptosporidium infection prevalence than that collected from River Nile which indicates an important public health problem and a potential risk of drainage canals in Egypt. ELISA showed higher prevalence of cryptosporidiosis than mZN for the total examined Clarias gariepinus fish. ELISA and PCR will be the "gold standard" where specificity and sensitivity will be very high, ensuring that undiagnosed Cryptosporidium infection does not occur. The phylogenetic analyses in this study confirmed this protozoal organism to be a novel detection of Cryptosporidium molnari in Clarias gariepinus fish.

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Shaapan, R.M., Abdel-Ghaffar, F.A., Varjabedian, K.G. et al. Prevalence and Molecular Epidemiology of Cryptosporidium Infection in Clarias gariepinus Fish in Egypt. Acta Parasit. 67, 437–445 (2022). https://doi.org/10.1007/s11686-021-00483-4

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