Molecular identification of paramphistomum SPP. in ruminants cut in Van municipality slaughterhouse

Since the morphological diagnosis of Paramphistomum species is insufficient by microscopic methods, molecular methods are needed for definitive diagnosis. In this study, it was aimed to identify by molecular methods of Paramphistomum spp in ruminants that were brought in Van Municipality slaughterhouse between March 2018 and September 2018. The study material consisted of adult Paramphistomum spp, collected from 100 infected ruminants (50 cattle and 50 sheep). DNA isolations of Paramphistomum parasites were made by commercial kit and then rDNA ITS2 gene region was amplified by PCR. The resulting PCR products were run on agarose gel and 399 bp bands were obtained. Amplicons were sequenced and compared with reference sequences in Genbank with BLAST. Genotypes of isolates were determined according to their complete or nearest similarities. Also, a phylogenetic tree was created with MEGA 7 program by using Neighbor-Joining Tree. In our study, bi-directional sequence analysis was performed from 10 PCR products obtained. As a result of the sequence analysis, 5 sheep and 3 cattle were identified as Paramphistomum leydeni, while 2 cattle were identified as Calicophoron daubneyi. In conclusion, previously diagnosed with of morphological methods C. daubneyi in Turkey which has become definite the diagnosis with molecular methods in Van province. Also the presence of the previously unreported P. leydeni was determined by molecular methods for the first time in Turkey.