Authors: GIREGOWDANAHALLI PUTTALAKSHMAMMA, UMED RAMANI, KRISHNA SINGH, AMRUTLAL PATEL, ABDUL PATEL, CHAITANYA JOSHI
Abstract: Paramphistomosis is widely prevalent in domestic farm animals, incurring heavy economic losses in several countries across the world. As the identification of these parasites based on morphological features is very difficult, molecular characterization is necessary to discriminate among different species. We performed polymorphic DNA fingerprint analysis of 3 different species of amphistomes collected from the rumen and bile ducts of buffaloes slaughtered at a local abattoir in Anand, Gujarat, India. The ruminal amphistomes were identified as Paramphistomum cervi and Gastrothylax indicus, while the hepatic amphistomes were identified as Gigantocotyle bathycotyle. High annealing temperature-random amplified polymorphic DNA (HAT-RAPD) analysis with 4 primers, OPA2, OPA4, OPA8, and OPA9, identified species-specific banding patterns for each species. The unique bands specific for species were sequenced, resulting in identification of 15 sequences, which included 6 sequences for G. bathycotyle, 3 sequences for G. indicus, and 6 sequences for P. cervi. The dendrogram analysis of RAPD bands revealed genetic distance of 0.5798 between G. bathycotyle and G. indicus, compared to a distance of 1.0986 and 0.9163 between G. bathycotyle and P. cervi and between G. indicus and P. cervi, respectively. The primers derived from sequenced RAPD amplicons generated one species-specific sequence characterized amplified region marker for the identification of G. indicus.
Keywords: Paramphistomes, DNA finger prints, dendrogram, random amplified polymorphic DNA, sequence characterized amplified region, buffalo
Full Text: PDF