Primary cell cultures from crustacea have been initiated since the 1960s, yet no permanent cell line is available. Primary cells have a limited proliferative capacity in culture due to cellular senescence, which is regulated by a group of dominant senescence genes. The aim of this research was to manipulate cell cycle regulation by transfecting Cherax quadricarinatus primary cells with oncogenes, in an effort to induce a permanent cell line. Human papillomaviruses (HPV) play a critical role in the formation of anogenital cancer. Research has demonstrated that the HPV-expressed E6 and E7 proteins function concomitantly to disrupt the p53 and retinoblastoma (Rb) tumor suppressor genes, regulators of the cell-cycle checkpoints at the first gap (G(1)) phase. HPV E6 and E7 genes were transfected into the C. quadricarinatus cells by lipofection. Successful transfection was demonstrated by the presence of oncogene messenger RNA by reverse transciptase polymerase chain reaction. At day 150, transfected cells still remain viable, although cell proliferation was stagnant. It may be that while transfection of the oncogenes was successful, no proliferation of the C. quadricarinatus cells was evident due to a lack of telomere maintenance.
Penaeus merguiensis densovirus (PmergDNV) is currently present on several Queensland prawn farms culturing Penaeus merguiensis. Densoviruses have been linked to mortality and stunting that has caused significant financial loss to prawn farms in Asia. A histopathological study for PmergDNV was initially undertaken to compare broodstock to grow out factors from 60 broodstock animals from each of 22 ponds. There was a significant negative correlation (r = -0.61) between the number of animals with PmergDNV lesions and healthy animals. Furthermore, a higher number of septic hepatopancreatic tubules was correlated (r = 0.48) to high PmergDNV loads. Hence, a polymerase chain reaction analysis of 10-day-old post-larvae (PL) was conducted to determine whether PmergDNV infection was resulting in production losses. An attributable risk analysis of PL from 190 ponds over a 2-year period revealed that 28-29% of ponds with below average survival will have at least average survival following the removal of or decreased levels of PmergDNV. P. merguiensis culture facilities in Queensland should have at least a 14.5% increase in production, equating to an increase of $2.25 million within the first year alone, following the removal or reduction of PmergDNV in their ponds. Hence, focussing efforts on prevention, better management practices and maintaining healthy stock should be of top priority.
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