Related Experiment Video
Updated: Mar 6, 2026
Mosquito-Associated Virus Isolation from Field-Collected Mosquitoes
Published on: August 31, 2022
Mosquito cells (C6/36) fail to support the complete replication of Penaeus merguiensis hepandensovirus
D Syahidah1, J Elliman2, C Constantinoiu2
1College of Public Health, Medical and Veterinary Science, James Cook University, Townsville, Australia; Institute of Marine Research and Development, Indonesia.
Abstract:
Mosquito cell lines (C6/36) were reported in the literature to support the propagation of Penaeus monodon hepandensovirus (PmoHDV). We aim to evaluate the susceptibility and viral propagation of P. merguiensis hepandensovirus (PmeHDV) which is ∼22% different to PmoHDV in Aedes albopictus cell line (C6/36). Cellular changes in the infected cell culture were detected. Vacuole formation was seen in both infected and uninfected cell cultures. The average number of disrupted cellular membranes in the infected cells (presumptive dead cells) was significantly higher than that of uninfected cells at passage two (F=9.749, d.f. 1, 22, p<0.05). Using a proliferation assay, light absorption of infected cells peaked at 2weeks post-infection (O.D.=0.27) but was significantly lower than that of the uninfected groups (O.D.=0.37) (F=6.879, d.f. 1, 94, p<0.05) suggesting hindered cell growth. PCR of the serial passages of the infected cell cultures indicated weak positive results for PmeHDV infection and TaqMan quantitative PCR confirmed that the average number of viral copies declined from 3.8×105 to 5.69×102 copies per μL and the mean of cycle times increased from 19.26 to 27.63. These results are interpreted to mean C6/36 allows the initial stage of PmeHDV replication, but the virus was incapable of using C6/36 for patent replication of its' virions.
Insights
The Aedes albopictus C6/36 cell line supports initial Penaeus merguiensis hepandensovirus (PmeHDV) replication but not sustained viral propagation. Viral loads declined significantly, indicating limited replication in this mosquito cell line.
Area of Science:
- Virology
- Cell Biology
- Invertebrate Pathology
Background:
- Mosquito cell lines, such as C6/36, have been utilized for the propagation of certain aquatic animal viruses.
- Penaeus monodon hepandensovirus (PmoHDV) has been previously shown to replicate in C6/36 cells.
Purpose of the Study:
- To investigate the susceptibility of the Aedes albopictus C6/36 cell line to Penaeus merguiensis hepandensovirus (PmeHDV).
- To evaluate the viral propagation and replication efficiency of PmeHDV in C6/36 cells.
Main Methods:
- Infection of C6/36 cells with PmeHDV.
- Monitoring cellular changes via microscopy.
- Assessing cell proliferation using a light absorption assay.
- Detecting viral presence and quantifying viral load using PCR and TaqMan quantitative PCR across serial passages.
Main Results:
- Significant increase in disrupted cellular membranes in infected cells compared to controls.
- Hindered cell growth observed in infected cultures, indicated by lower light absorption.
- Weak positive PCR results for PmeHDV, with a significant decline in viral copies (3.8×10^5 to 5.69×10^2 copies/μL) and an increase in cycle times across passages.
Conclusions:
- The C6/36 cell line supports the initial stages of PmeHDV replication.
- PmeHDV is incapable of patent replication and sustained propagation in the C6/36 cell line.
- This study highlights the limitations of using C6/36 cells for PmeHDV research.
