Development of DNA barcode approach and its comparison with classical microscopy: Assessment of nematodes in Punjab Urial (Ovis vignei punjabiensis) Population in Kalabagh Game Reserve

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Authors

A Bajwa1; S Islam1
1 University of Veterinary and Animal Sciences, Pakistan

Discussion

Punjab Urial (Ovis vignei punjabiensis) is a wild sheep of Pakistan, having a status of vulnerable species by IUCN. The major threats include habitat loss and poaching, leading to a severe decline in its population. In addition, nematode infections may also have compromised its survival. However, Punjab Urial has hardly been studied to explore the gastrointestinal nematode infections. In this context, we designed a study to determine the presence of nematodes in Punjab Urial from Kala Bagh Game Reserve, District Mianwali. A new DNA barcoding approach was developed exploring ITS-I as a target region, with a primer pair designed to amplify frequently reported nematode species of small ruminants. The results of DNA barcoding were compared with those of classical microscopy. DNA barcoding revealed higher prevalence of Haemonchus contortus (73.91% vs 35.86), Trichuris ovis (16.30% vs 6.52%) and Trichostrongylus axei (3.26% vs 0%) in Punjab Urial as compared with faecal egg examination. The study demonstrates the new DNA barcoding approach as a robust tool to detect nematode parasites from faecal samples of Punjab Urial. Our approach also mitigates problems associated with microscopic recognition and can be executed to detect nematode infections in wild and domestic hosts for surveillance and population conservation. 
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