Molecular Characterization of Plasmodium relictum in Four Common Bird Species in India

ABSTRACT The increasing emergence of wildlife diseases with the possibility to ecological threats as well as domestic animals and human health has prompted the importance of understanding disease dynamics and associated risks in biological conservation. The present study was undertaken from North Eastern part of India from January, 2017 to March, 2018 to identify and molecularly characterize Plasmodium relictum based on cyt b gene in various species of wild birds (Upupa epops, Passer domesticus, Pycnonotus cafer, Bubulcus ibis). The birds were captured by netting system. After blood was collected from wing veins, birds were released from the cages. Blood samples were examined after staining with Giemsa stain. The positive samples were used for amplification of cyt b gene of Plasmodium relictum. Cloning and sequencing of the amplified products of each samples was performed separately. Out of 120 birds examined, 25 were found positive for P. relictum based on morphology and subsequently confirmed by PCR which selectively amplified 712bp of P. relictum. Based on sequences and phylogenetic analysis of cyt b gene, 8 isolates of P. relictum were identified. The obtained complete nucleotide sequences of cyt b gene from P. relicum revealed100% identity among themselves while the other sequences registered in Gen Bank showed 95% to 99% similarity. In the phylogenetic analysis all the isolates of P. relictum formed a separate clade with high bootstrap values. It can be inferred from the study that P. relictum is fairly common in wild birds and the cyt b gene is highly conserved among different isolates. It seems that cytb could provide suitable genetic markers for discrimination and genetic characterization of P. relictum. This is the first attempt of genetic characterization of P. relictum from India.