Discovery of an Australian Chelonia mydas papillomavirus via green turtle primary cell culture and qPCR

Mashkour, Narges, Maclaine, Alicia, Burgess, Graham W., and Ariel, Ellen (2018) Discovery of an Australian Chelonia mydas papillomavirus via green turtle primary cell culture and qPCR. Journal of Virological Methods, 258. pp. 13-23.

[img] PDF (Published Version) - Published Version
Restricted to Repository staff only

View at Publisher Website: https://doi.org/10.1016/j.jviromet.2018....
1


Abstract

The number of reptilian viruses detected are continuously increasing due to improvements and developments of new diagnostic techniques. In this case we used primary cell culture and qPCR to describe the first Australian Chelonia mydas papillomavirus. Commercial chelonian cell lines are limited to one cell line from a terrestrial turtle (Terrapene Carolina). To establish primary cultures from green turtles (Chelonia mydas), turtle eggs were collected from Heron Island, Queensland, Australia. From day 35 of incubation at 29 degrees, the embryos were harvested to establish primary cultures. The primary cell cultures were grown in Dulbecco's Modified Eagle Medium, 90% and foetal bovine serum, 10%. The cells became uniformly fibroblastic-shaped after 15 passages. The growth rate resembled that of cells originating from other cold-blooded animals and the average doubling time was similar to 5 days from the 20th passage. Karyotyping and molecular analysis of mitochondrial DNA D-loop gene were carried out for cell authentication. The primary cell cultures were screened to exclude mycoplasma contamination. Two primary cell lineages were found to be susceptible to Bohle iridovirus. The primary cell cultures were used to screen samples from green turtles foraging along the East Coast of Queensland for the presence of viruses. Homogenates from eight skin tumour samples caused cytopathic effects and were confirmed by qPCR to be infected with papillomavirus.

Item ID: 54584
Item Type: Article (Research - C1)
ISSN: 1879-0984
Keywords: Green turtle, primary cell culture, qPCR, papillomavirus
Date Deposited: 18 Jul 2018 07:43
FoR Codes: 07 AGRICULTURAL AND VETERINARY SCIENCES > 0707 Veterinary Sciences > 070712 Veterinary Virology @ 100%
SEO Codes: 96 ENVIRONMENT > 9604 Control of Pests, Diseases and Exotic Species > 960407 Control of Pests, Diseases and Exotic Species in Marine Environments @ 100%
More Statistics

Actions (Repository Staff Only)

Item Control Page Item Control Page