Morphological and Molecular Identification of Paramphistomum epiclitum from Buffaloes in Pakistan



Little is known about the genetic and morphological characters of Paramphistomum epiclitum. For the first time in Pakistan, adult flukes were morphologically characterized and the sequence variation in the nuclear ribosomal DNA (rDNA) region, including the first internal transcribed spacers (ITS1) and the 5.8S gene of the Paramphistomum epiclitum were studied.


Adult amphistomes were examined by light microscopy and sequences of ITS1 and 5.88S rDNA genes were obtained.


Twenty adult flukes were measured, 13.17 ± 1.19 mm in length and 5.28 ± 1.34 mm in width. Seventeen adult flukes were sequenced and high sequence variability was observed in 5′ end of ITS1 region. The 5.8S and 3′ end of ITS1 sequences had 100% identity among the samples. A comparative analysis revealed that different types and numbers of repeats were found within each ITS1 region. The 3′ end of ITS1 region from P. epiclitum showed 98% homology with P. cervi from China and formed a subclade with genetic distance of 0.1663. The 5.8S gene showed 100% identity within Paramphistomidae family and formed a sub-clade with P. epiclitum, P. leydeni, P. cervi and Cotylophoron cotylophorum species isolated from China, India and Uruguay.


This work provides new information on morphological identity and genetics of P. epiclitum from Pakistan.

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The research work presented in this article was conducted with internal funds of Quaid-i-Azam University, Islamabad. We are grateful to the Emeritus Professor Dr James E Miller Department of Pathobiological Sciences, School of Veterinary Medicine, Louisiana State University, USA for proofreading and English editing of the manuscript.

Author information




KA designed the study. IK, SS, SA and MK performed the experiment. KA, SF conducted the literature search, data analysis and manuscript preparation, advised on methods and interpretation of findings. KA and SF reviewed the manuscript. All authors participated in the study and concur with the submission and subsequent revisions submitted by the corresponding author.

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Correspondence to Kiran Afshan.

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The author(s) declare no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Ethical approval

The study was conducted by following the guideline approved by the Ethical Committee of Quaid-i-Azam University Islamabad, Pakistan and has therefore been performed in accordance with the ethical standards laid down in the 1964 Declaration of Helsinki and its later amendments. All animals used in the study were slaughtered for other purposes to fulfill the protein demand of the local population.

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Khan, I., Afshan, K., Shah, S. et al. Morphological and Molecular Identification of Paramphistomum epiclitum from Buffaloes in Pakistan. Acta Parasit. 65, 225–236 (2020).

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  • Paramphistomatidae
  • ITS1
  • 5.8S
  • Morphological characterization
  • Molecular characterization
  • Pakistan