How to set up a aquaculture pathogen challenge laboratory?

While there are many applicable protocols, controls, and facility design aspects that can be adapted from BSL-2 and BSL-3 labs, aquatic pathogen experimentation provides some unique challenges. Much of the setup will depend of course on the aquaculture species and pathogens of interest. As well, the scale of challenges will be important to consider; single subject replication vs group replication; juveniles vs adults, etc. Design-wise, isolation of all potentially exposed equipment, gear, etc. should be ensured and the facility hosting challenge trials should be physically separated from production facilities. Many trials adopt standard toxicological study designs, which allow challenge trials to be done in standard BSL-2 labs. Some further considerations:

Mimicking production conditions (group replicates) in many cases requires mechanical filtration, biofiltration, and aeration. As challenge trials typically require high replication, filtration processes should be simple, self contained (for ease of renewal, cleaning, etc.), and integrated into an effective flow system. Aeration can be a part of the filtration process (e.g. airlift), however, the production of aerosols (think cross contamination and containment) and foams (especially with high organic loads and mortality in waters, even more so in brackish/marine water). Aerosols can be captured within tanks if a negative pressure exhaust system is integrated with tanks or chambers, but opening tanks for feeding or removing animals can expose lab workers, so proper PPE should be worn. Keep EtOH around in spray bottles everywhere.

Observation is important. Many animal care protocols demand euthanization at the onset of symptoms, sometimes at morbidity. Cannibalism can be an etiological effect, but if you are controlling for routes (i.e. by feed or injection), moribund individuals should immediately be removed from the tanks. Having someone sit in the lab and watch tanks is likely undesirable, and repeated entry/exit increases the risk for breakdown in containment protocols. A camera system can be very advantageous. Hand written data can be recorded on plastic films.

Provide more room than necessary for logistics and cleaning. Implement a draining and pumping system that is easy to use, minimizes dripping, and can be operated by one person. Moving high volumes of contaminated water around can pose a hazard. Post-treatment of contaminated water should be in the standard protocols (i.e. chlorine dosing tanks before cleaning. All PPE, clothing, instruments, etc. should be removed in an intermediate area, before exiting the controlled areas.

Environmental controls should include filtration of air (intake and exhaust) and temperature regulation; try to eliminate direct sunlight and control water temperatures directly instead of the room air temperature (e.g. modular heat exchange system). The challenge area should be negative pressure. Floors should be sealed along the walls, and floor drains should be in a treated waste stream. Have a separate dry room for feeds (or in a refrigerator).

Depending on the pathogen, animal disposal can go several directions. Many times it is easiest to hire a company to incinerate or render, but they must be kept cool until transfer, and if rendered, it is very important to assure the process neutralizes the pathogen. Transport of animals in and out of the facility should be done in sealed containers. Keep aware that when you open a challenge laboratory, you can easily be a target of accusation when outbreaks occur, regardless if they are related to your activities. So having documented, well-established protocols and controls will be key.

Barry C Smith

As Daniel wrote, I will add to: Remember containment and decontamination. Even if the local laws do not require absolute containment, they may in the future. Also, you do not want to be accused of a release.

Noori Abdul-nabi Nasir

It is good work to do to meet the rapid of increasing fish farming at the present time.

Gen Hua Yue

Thank you very much for your answers. They are very useful to us.

What is your major research area?

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