Molecular characterization of Raillietina isolates from the gastrointestinal tract of free range chickens (Gallus Gallus domesticus) from the southern region of Zimbabwe using the 18S rDNA gene.

Abstract

Parasitic diseases are among the major constraints of poultry production. The common internal parasitic infections occurring in poultry include cestodes and other gastrointestinal helminths. The aim of this study was to characterize Raillietina spp. from the gastrointestinal tract of free-range chickens using 18S rDNA genes. This was achieved through the DNA extraction from tapeworms isolated from the gastrointestinal tract of routine slaughtered free-range chickens using 18S and amplifying the partial 18S rDNA genes using PCR. A total of 34 intestine samples were collected from 9 different areas in Matabeleland region and tapeworms were isolated for microscopic examination and morphological characterization. All 34 chickens examined had tapeworms and morphological characterization confirmed those used in the study as belonging to the Raillietina group. DNA extraction was then successfully carried out for 9 cestode isolates followed by the PCR amplification of the 18S genes. The products were sequenced and 4 of the 9 isolates were positively identified using BLAST as Raillietina tunetensis and the other 5 were only identified as Raillietina spp. These sequences were then aligned with other known sequences of Raillietina isolates from GenBank and a phylogenetic tree was constructed using the neighbor-joining method and distance computed using the maximum composite method. The phylogenetic tree showed that three of the unidentified Raillietina species are in the neighboring position with Raillietina sonini whilst the other two are in the neighboring position with Raillietina tetragona. This is the first record of R. tunetensis in Gallus Gallus domesticus from Zimbabwe. This clustering on the phylogenetic tree however, did not differentiate samples according to geographical location showing that this tool can be used to infer phylogenetic data for speciation.