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Stability of an infectious pancreatic necrosis virus (IPNV) isolate stored under different laboratory conditions
S H Mortensen1, R K Nilsen, B Hjeltnes
1Institute of Marine Research, Department of Aquaculture, Bergen, Norway.
Abstract:
An infectious pancreatic necrosis virus (IPNV) belonging to the Sp serotype, isolated from scallops Pecten maximus, was propagated and suspended in sterile water and cell culture medium with different salinities and incubated at temperatures ranging from -80 to 40 degrees C. Virus stability was examined by measuring virus titers under different storage conditions. Virus titers were also measured after repeated freezing and thawing, and in incubated sterile filtered scallop hepatopancreas, haemolymph and crystalline style samples, and salmon Salmo salar kidney homogenate. The virus was stable under most storage conditions. Temperatures ranging from -80 to +20 degrees C, as well as salinities from 0 to 40/1000, did not seem to influence the stability of the virus. A reduction was observed above 20 degrees C. Each freezing and thawing procedure resulted in a reduction of the virus titer. This reduction was larger at -80 than at -20 degrees C. The IPNV persisted for a long period in sterile filtered scallop haemolymph, dissolved crystalline style and hepatopancreas. In kidney homogenate from IPNV-infected salmon the virus titers were reduced at least 10-fold during the first day of incubation at all temperatures tested. When virus was propagated in cell culture and subsequently mixed with non-infected salmon kidney homogenate, the virus proved more persistent. Our results illustrate the importance of rapid and standardised laboratory processing of potentially virus-containing tissue samples, and are relevant when considering laboratory storage of samples containing IPNV.
Insights
Infectious pancreatic necrosis virus (IPNV) remains stable across various temperatures and salinities. However, repeated freezing and thawing reduce virus titers, impacting sample storage strategies for IPNV detection.
Area of Science:
- Aquatic Animal Health
- Virology
- Molecular Biology
Background:
- Infectious pancreatic necrosis virus (IPNV) is a significant pathogen affecting aquatic species.
- Understanding IPNV stability is crucial for accurate diagnostics and disease management.
Purpose of the Study:
- To evaluate the stability of infectious pancreatic necrosis virus (IPNV) under various storage and processing conditions.
- To determine optimal laboratory practices for handling IPNV-infected samples.
Main Methods:
- IPNV (Sp serotype) from scallops (Pecten maximus) was tested for stability.
- Virus titers were measured after incubation at temperatures from -80°C to 40°C and salinities from 0 to 40/1000.
- Stability was assessed after repeated freezing/thawing and in various tissue homogenates (scallop and salmon).
Main Results:
- IPNV remained stable at temperatures between -80°C and +20°C and salinities of 0-40/1000.
- Virus titers decreased significantly above 20°C and with each freeze-thaw cycle, more so at -80°C than -20°C.
- IPNV showed prolonged persistence in sterile scallop haemolymph, crystalline style, and hepatopancreas.
Conclusions:
- IPNV stability is influenced by temperature and freeze-thaw cycles.
- Rapid and standardized laboratory processing is essential for reliable IPNV detection in clinical samples.
- Findings inform best practices for sample collection and storage in IPNV surveillance programs.