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Published on: August 31, 2022
In vitro propagation of infectious myonecrosis virus in C6/36 mosquito cell line
S Santhosh Kumar1, S Sivakumar1, S Abdul Majeed1
1Aquatic Animal Health Laboratory, C. Abdul Hakeem College, Ranipet, India.
Abstract:
Infectious myonecrosis (IMN) is an important shrimp viral disease caused by infectious myonecrosis virus (IMNV). Based on previous reports, an attempt was made to propagate IMNV in apparently healthy C6/36 subclone of Aedes albopictus cell line. The confirmatory assays such as RT-PCR, real-time PCR and bioassay revealed that C6/36 cells were found to be susceptible to IMNV and these cells could be used easily for isolation and propagation of IMNV. The results of real-time PCR assay showed that a lower CT value of 22.25 in IMNV-infected cells was obtained on 10 day post-infection (d p.i.), whereas the higher CT value of 35.21 was obtained in IMNV-infected cells on 2 d p.i. There is no significant difference between CT values of IMNV production in vitro using C6/36 cell line and in vivo using shrimp. The IMNV propagated in C6/36 cells is capable of infecting shrimp and caused 100% mortality in shrimp. Clinical signs observed in shrimp injected with IMNV propagated in C6/36 cell line were found to be similar to naturally infected shrimp.
Insights
Infectious myonecrosis virus (IMNV) can be effectively propagated in Aedes albopictus C6/36 cells. This cell line offers a reliable method for IMNV isolation and propagation, crucial for studying this significant shrimp disease.
Area of Science:
- Aquaculture
- Virology
- Cell Biology
Background:
- Infectious myonecrosis (IMN) is a severe viral disease impacting shrimp aquaculture globally.
- The infectious myonecrosis virus (IMNV) is the causative agent responsible for this devastating disease.
Purpose of the Study:
- To investigate the susceptibility of the Aedes albopictus C6/36 cell line to IMNV.
- To evaluate the potential of C6/36 cells for the isolation and propagation of IMNV.
Main Methods:
- Propagation of IMNV in C6/36 cells.
- Confirmation of infection using RT-PCR and real-time PCR.
- Bioassays to assess the infectivity of propagated virus in shrimp.
Main Results:
- C6/36 cells demonstrated susceptibility to IMNV infection.
- Real-time PCR showed viral replication with CT values comparable to in vivo infections.
- IMNV propagated in C6/36 cells caused 100% mortality in shrimp with similar clinical signs.
Conclusions:
- The Aedes albopictus C6/36 cell line is a suitable host for IMNV isolation and propagation.
- This in vitro system provides a valuable tool for IMNV research and potentially for developing control strategies.
