Molecular characterization and phylogenetic analyses of Fasciola gigantica of buffaloes and goats in Punjab, Pakistan. (June 2021) - Rehman, Zia Ur
Abstract: Fasciola gigantica is considered to be a major pathogen causing fasciolosis in the Indian subcontinent, resulting in production losses of millions of dollars in the livestock industry. Understading the dispersal origin and the patterns of spread of F. gigantica is important. A total of 53 Fasciola flukes collected from buffaloes and goats in Punjab, Pakistan between 2017 and 2018 were identified as F. gigantica based on the multiplex PCR for the phosphoenolpyruvate carboxykinase ( pepck ) and the PCR-restriction fragment length polymorphism (RFLP) for DNA polymerase delta ( pold ). A significant genetic difference between F. gigantica from buffaloes and goats was indicated by the genetic analyses of mitochondrial markers, NADH dehydrogenase subunit 1 ( nad1 ) and cytochrome C oxidase subunit 1 ( cox1 ). Phylogenetic analysis of the seventeen nad1 haplotypes of F. gigantica from Pakistan with those in neighbouring countries of the Indian subcontinent revealed that all the haplotypes identified in Pakistan were clustered in haplogroup A. fasciola gigantica with the eight haplotypes might be expanded in Pakistan from Indian origin, along with the migration of the domestic animals, since they were related to Indian haplotypes . In contrast, the remaining nine haplotypes were not shared with any neighbouring countries, suggesting independent origin, probably from neighbouring Middle East countries. However, cautious interpretation is required due to the very limited samples size of this study. Our study provides a proof of concept for a method that could be used to investigate the epidemiology of F. gigantica . Graphical abstract: Unlabelled Image Highlights: Pakistani Fasciola flukes were identified as F. gigantica by nuclear pepck and pold . Mitochondrial nad1 and cox1 were used for phylogenetic characterizations. Genetic difference between F. gigantica from buffaloes and goats was suggested. Two origins of F. gigantica in Pakistan were indicated by the network analysis.