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Morphological and molecular characterisation of common amphistome species from cattle of South India

Shameem H, Devada K, Lakshmanan B, Joseph S, Sabu L, Usha AP

Bovine amphistomosis is a highly neglected snail borne trematode disease causing great economic loss to dairy farmers. The aim of the present study was morphological identification of different amphistome species and molecular characterization of the most common amphistomes prevalent in the study area. Among the ten different species of amphistomes identified in the present study four common species namely, F. c o b b o l d i, F. e l o n g a t u s, G. c r u m e n if e r and Paramphistomum spp. were genetically characterized by amplifying second internal transcribed spacer sequence flanking 5.8S and 28S partial ribosomal gene sequences (ITS-2+) which produced amplicons of 494 bp, 503 bp, 514 bp and 494 bp respectively. The nucleotide sequences of four species of amphistomes F. cobboldi, F. elongatus, G. crumenifer and Paramphistomum spp . obtained in the present study is the first report from Kerala (India) which provides the primary sequence data of the rDNA ITS-2+ region of the amphistome flukes. Nucleotide sequence analysis of the four species revealed eight nucleotide differences between the sequences of pouched amphistomes ( F. cobboldi, F. elongatus and G. crumenifer ) and unpouched ( Paramphistome spp.). Nucleotide sequence analysis and in silico restriction map analysis of four amphistomes suggest the possibility of ITS-2+ region as a useful genetic marker for species identification.