Molecular Characterization of Paramphistomum epiclitum (Trematoda: Paramphistomidae) Based on First Internal Transcribed Spacer (ITS-1) of Ribosomal DNA from Buffaloes of Peshawar, Pakistan

Abstract

Objectives: Helminthes parasitism is the main problem that limits animal productivity. Epidemiological pattern of disease would provide a base for premediated control of disease. The objective of the present study is to characterize the Paramphistomum epiclitum on the base of internal transcribed spacer 1 region and to establish phylogenetic analysis from buffaloes of Khyber Pakhtunkhwa, Pakistan and to investigate intraspecific and interspecific variation with other trematodes. Methods: Paramphistomum collected from rumen of infected buffloes (n=50). DNA was extracted through phenol chloroform method and amplified through PCR. PCR product was purified and sequenced. Results were analyzed through BLAST and CLUSTALW. Phylogenetic analysis was performed and genetic distance was calculated through MEGA version 7 by maximum likelihood method. Results: The study revealed existence of intraspecific variation with other trematodes. Maximum homology was observed with Paramphistomum cervi of China that is 97% and with Gastrodiscoides hominis of India with similarity index of 93%. Chiorchis febaceaus of Germany deviate from current study and shows homology of 83%. The minimum genetic distance of 0.1663 was observed with Paramphistomum cervi and maximum distance of 2.4756 was observed in case of Choirchis febaceaus of Germany. Conclusion: It is concluded from present studies that P. epicilitum of current study showed resemblance with trematodes of geographically linked countries. Phylogenetic tree and pairwise genetic difference showed strong affiliation with Paramphitomum cervi of China and Gastrodiscoides hominis of India. Chiorchis febaceaus of Germany show deviation from current specie.