Molecular detection and genetic diversity of avian haemosporidia (Plasmodium and Haemoproteus) infecting red jungle fowls (Gallus gallus spadiceus) and domestic poultry in peninsular, Malaysia

AbstractThis study constitutes the first attempt to characterized Plasmodium and Haemoproteus affecting Gallus gallus complex (Red Jungle fowl, village chickens and broilers) (Gallus gallus spadiceus, Gallus gallus domesticus and Gallus gallus domesticus respectively) in peninsular, Malaysia. A total of 713 blood samples were collected from 713 G. gallus complex in peninsular, Malaysia in different farms and villages across peninsular, Malaysia. From the total of 713 blood samples collected, 241 samples were from red jungle fowls, 322 were from village chickens and 150 from broilers. All the samples were subjected to both microscopy and molecular detection of plasmodium and Haemoproteus. The prevalence of Plasmodium was found to be 4.6% and that of Haemoproteus was found to be 1.1% microscopically. Using molecular detection, the PCR screening reveal the prevalence of Plasmodium to be 30.6% and that of Haemoproteus was found to be 10.7%. All the positive samples by PCR screening which target the partial segment of the cytochrome b gene of the parasite with 500 bp (478 bp without primers) were cloned; ten colonies were picked after PCR confirmation of the colonies. Plasmids were extracted from the positive PCR screened colonies and sequence. The sequence results were manually edited, and National Center for Biotechnology Information (NCBI) Basic Local Alignment Search Tool (BLAST) analysis was done to compare with the sequence from the previous studies for molecular characterization. Our result reconfirmed the morphological characterization of Plasmodium gallinaceum, P.parasitese and Haemoproteus sp. in Gallus gallus complex in peninsular, Malaysia by molecular characterization and phylogenetic analysis of partial 500 bp (478 bp without primers) segment of the cytochrome b gene of the parasites.

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