Morphological and molecular characterization of Calicophoron raja (Näsmark, 1937) collected from wild Bovidae in South Africa

Paramphistomes, commonly known as rumen flukes, are digenean parasites that infect ruminants. Accurate morphological identification of paramphistome species is challenging and often neglected. For instance, it requires sagittal midline sections of adult flukes, which are difficult to prepare. Therefore, the majority of the genetic information on paramphistomes found in the International Nucleotide Sequence Database is not supported by morphological descriptions, and the DNA barcodes of paramphistome species remain unreliable. In the present study, both morphological and molecular characterizations were simultaneously performed to ensure the reliability of the DNA information for the paramphistome species Calichophoron raja (Näsmark, 1937). The morphological characteristics of the sagittal and horizontal sections of adult flukes from a black wildebeest (Connochaetes gnou) and a waterbuck (Kobus ellipsiprymnus) in South Africa were identical to those previously described for Ca. raja. Additionally, this study represents a new host record of the species from Co. gnou. All sequences of the internal transcribed spacer 2 region of ribosomal DNA were 100% identical among the 18 flukes analyzed in the present study. A single nucleotide mutation was observed between Ca. raja in this study and Ca. raja detected in domestic ruminants in Kenya.