A Molecular Method for Identifying Calicophoron Infecting Cattle in South Africa
A molecular method for identifying Calicophoron infecting cattle in South Africa
Lavinia Perumal
School of Biological Sciences and Conservation, University of KwaZulu-Natal, Westville, Durban. Email:209512772@ukzn.ac.za
Abstract
Calicophoron species were collected from the ruminants of cattle from a Kokstad abattoir in South Africa. The second internal transcribed spacer (ITS-2) of the ribosomal DNA had been used as the genetic marker in this molecular study. In an attempt to identify as well as distinguish Calicophoron, the sequences of the ITS-2 region of various species obtained from the experimental samples were compared to each other and to the sequences of related species obtained from the NCBI Genbank.DNA was isolated from individual flukes collected from six cows, PCR techniques amplified segments of the DNA and allowed for purification and sequencing. Sequence analysis comparisons provided a basis for the phylogenetic tree (neighbour-joining), this indicated that there were two species found, one unnamed and the other C.daubneyi and of the six cows, two were co-infected .B.forskali acted as an intermediate host for the unnamed Calicophoron species, the sequences of four samples were identical to the unnamed Calicophoron species isolated from cercaria of B.forskali. Intestinal paramphistomes are found to be more common in cattle than that of which is indicated by literature, which may be due to difficulty in diagnosis, although sequence comparisons of the ITS-2 regions increased the ability to identify and distinguish Calicophoron.
Introduction
The disease caused by infestation of ruminants of various hosts with stomach flukes is known as paramphistomosis. The paramphistomidae family consists of various genera, one of which is Calicophoron. This family is characterized by the absence of oral suckers and an acetabulum that is positioned close to the posterior end in adults (Fischoeder, 1901). The disease is known to affect cattle and sheep;
Lavinia Perumal
School of Biological Sciences and Conservation, University of KwaZulu-Natal, Westville, Durban. Email:209512772@ukzn.ac.za
Abstract
Calicophoron species were collected from the ruminants of cattle from a Kokstad abattoir in South Africa. The second internal transcribed spacer (ITS-2) of the ribosomal DNA had been used as the genetic marker in this molecular study. In an attempt to identify as well as distinguish Calicophoron, the sequences of the ITS-2 region of various species obtained from the experimental samples were compared to each other and to the sequences of related species obtained from the NCBI Genbank.DNA was isolated from individual flukes collected from six cows, PCR techniques amplified segments of the DNA and allowed for purification and sequencing. Sequence analysis comparisons provided a basis for the phylogenetic tree (neighbour-joining), this indicated that there were two species found, one unnamed and the other C.daubneyi and of the six cows, two were co-infected .B.forskali acted as an intermediate host for the unnamed Calicophoron species, the sequences of four samples were identical to the unnamed Calicophoron species isolated from cercaria of B.forskali. Intestinal paramphistomes are found to be more common in cattle than that of which is indicated by literature, which may be due to difficulty in diagnosis, although sequence comparisons of the ITS-2 regions increased the ability to identify and distinguish Calicophoron.
Introduction
The disease caused by infestation of ruminants of various hosts with stomach flukes is known as paramphistomosis. The paramphistomidae family consists of various genera, one of which is Calicophoron. This family is characterized by the absence of oral suckers and an acetabulum that is positioned close to the posterior end in adults (Fischoeder, 1901). The disease is known to affect cattle and sheep;
References: Dinnik, J.A., 1964a.Intestinal paramphistomiasis and P. microbothriumFischoeder in Africa. Bull. Epizoot. Dis. Afr. 12, 439–454. Dube S,Masanganise K.E and Dube C., 2010 . Studies on paramphistomes infecting goats and sheep from Gwanda District in Zimbabwe . ZJST . 5 , 55 - 64 Fischoeder F.,1901 24, Ilha, M.R., Loretti, A.P., Reis, A.C., 2005 Itagaki, T., Tsumagari, N., Tsutsumi, K., Chinone, S., 2003. Discrimination of three amphistome species by PCR-RFLP based on rDNA ITS2 markers.J. Vet. Med. Sci. 65, 931–933. Jones, A., 1990. Techniques for hand-sectioning thick- bodied Platyhelminths.Syst. Parasitol. 15, 211–218. Khan, U.J., Tanveer, A., Maqbool, A., Masood, S., 2008. Epidemiological studies of paramphistomosis in cattle. Vet. Arhiv 78, 243–251. Lofty W.M,, Brant S.V, Ashmawy K.I, Devkota R.Mkoji G.M, Loker E.S.,2010. A molecular approach for identification of paramphistomes from Africa and Asia. Veterinary Parasitology 174,234-240 Näsmark, K.E., 1937 Nolan M.J and Cribb T.H., 2005. The use and implications of ribosomal DNA sequencing for the discrimination of digenean species.AdvParasitol 60,101–163. Olson, P.D., Cribb, T.H., Tkach, V.V., Bray, R.A., Littlewood, D.T.J., 2003. Phylogeny and classification of the Digenea (Platyhelminthes: Trematoda).Int. J. Parasitol. 33, 733–755. Phiri A.M, Chota A, Phiri I.K., 2007. Seasonal pattern of bovine amphistomosis in traditionally reared cattle in the Kafue and Zambezi catchment areas of Zambia. Trop Anim Health Prod 39,97–102. Rangel R L.J.,Brahms A, Aguilar J., 2003. Seasonal trends of Paramphistomumcervi in Tabasco, Mexico. Veterinary Parasitology 116,217–222. Rolfe, P.F., Boray, J.C., Nichols, P. and Collins, G.H., 1991. Epidemiology ofParamphistomosis in cattle.International Journal for Parasitology.7, 813 – 819. Sanabria, R.E.F., Romero, J.R., 2008. Review and update of paramphistomosis.Helminthologia 45, 64–68. Sey, O., Prasitirat, P., Romratanapun, S. &Mohkaew, K., 1997. Morphological studies and identification of rumen flukes of cattle in Thailand. Rivista di Parassitologia 2, 247–256.