IACUC Guideline 10 - Aquatic Guidelines
Investigators who will maintain aquatic species at Penn State University for use in research, teaching or testing are required to establish written standard operating procedures (SOPs) which describe routine care and monitoring. SOPs along with records of routine monitoring must be readily available within the aquatic facility at all times (either posted or maintained in a notebook). New or significantly revised SOPs should be prepared in consultation with and/or approved by a qualified representative of the Animal Resource Program. The IACUC will assess the effectiveness of procedures during semiannual inspections.
An arrangement must be established to insure that all animals and their housing environment are assessed on a daily basis.
Routine and emergency contact information must be clearly posted at the facility entrance.
Standard Operating Procedures:
The scope and complexity of the SOP may vary greatly depending on the species and type of housing. The list below provides suggestions for information to include. In some cases additional information may be necessary; in others only a few of the points below may need to be addressed.
- Procedures for daily observations
- Procedures for cleaning aquariums (or other housing unit)
- Procedures for water conditioning
- Diet and general feeding plan
- Record keeping procedures
- Frequency of:
- Water quality monitoring
- Water change
- Room sanitation
- Lighting schedule
- Water temperature
- Procedures for monitoring water quality
- Parameters to monitor (temperature, pH, dissolved oxygen, ammonia, nitrites, nitrates, etc.)
- Monitoring frequency
- Procedures for room sanitation
- Filter maintenance procedures
- UV light change
The principle investigator is responsible for insuring that his or her investigative staff and students have received adequate training and are competent to perform their assigned duties. All personnel must also complete the on-line course in animal care and use offered by the IACUC.
Aquatic Animal Health:
Tanks are to be checked daily and any dead or ill fish removed, recorded, and reported. Tanks in which diseased fish are noted must be reported to the veterinary personnel at the Centralized Biological Laboratory (CBL). Methods of prophylactics, diagnoses, control, and treatment of the disease and injuries follow currently accepted practices for aquatic vertebrates.
Anesthesia, Analgesia, Euthanasia:
Appropriate use of anesthetics and analgesics should be as described in the IACUC protocol. Methods for euthanasia must be consistent with the AVMA Guidelines on Euthanasia. Please consult with an Animal Resource Program veterinarian for additional guidance.
In general, unless the fishes are held at or near their optimum lethal temperature, the water temperature should be elevated 5°C and treated with iodized salt at a concentration of 3 g/l. Salt treatment is effective against fungi, protozoans, and other eukaryotes, as well as bacterial infections. Most freshwater fishes can tolerate concentrations as high as 10% and cichlids can tolerate concentrations as high as 60%, which is roughly twice the saline concentration as seawater. Salt treatment also has the advantage in that the salt is completely purged from the fish's body, once they are returned to freshwater. If the disease cannot be cured with salt treatment, then tissues are given to CBL for analysis. Fishes will then be treated with the recommended antibiotic. If the disease prevails, then the fish are destroyed, the tank cleaned with dishwashing liquid, and soaked in a Clorox solution for 24 hours. Any nets used to capture diseased fishes are rinsed in bleach immediately after being used.
Rooms also need to have a net disinfection system. Such a system should provide for ONE-TIME-USE of nets. A supply of sanitized or autoclaved nets are to be made available in a "clean bucket". Following each procedure, one-time-use nets are placed in a "dirty bucket" for cleaning. Nets are never to be used among tanks or experiments.
Rooms need to have an area (centralized in larger facilities) where tanks can be cleaned, sanitized, and dried. Holding areas and tanks need to be kept clean. Care should be taken not to contaminate water from one tank with that from another. It is especially important to use clean nets for each application.
The water source is carefully considered for all facilities. Deep wells are the best source of fresh water because they contain fewer infectious agents and/or toxic chemicals such as sewage or agricultural chemicals. Municipal tap water, in most areas, must be treated for the removal of chlorine/chloramine before use. Water quality levels are monitored and maintained at levels appropriate for specific organisms. Each individual principal investigator will develop standard operating procedures for the organisms being held in her/his facility.
Feed is to be wholesome, palatable, free of contaminants, and kept in sealed containers. Cleaning agents are to be stored away from the food. Detailed information about the diet, including storage is to be provided in the SOP.
It is almost impossible to provide stocking rates, even with regard to a particular proposal, because of the differences in sizes of fish and sizes of aquaria. A density of more than 1.5 cm of fish per liter of water should not be exceeded, unless justification for higher densities is part of the experimental design and documented in a specific proposal. Each tank must be identified with an identification number assigned by the IACUC.
General Features of Aquatic Animal Facilities:
Wall coverings, and floor treatments, door thresholds, and to a lesser extent ceilings (depending upon height) are of materials impervious to water or made resistant as necessary. Doors to the holding rooms must have thresholds that prevent water escape.
Copper piping and lead-based solders should be avoided. Rooms should be provided with adequate facility-wide drainage, which requires drains in several locations. Steeply angled slopes to central floor drains must be avoided because it creates unstable footing for heavy aquarium ranks.
Floor drains should be present in all facilities. Specialized drains are an important consideration. Water with infectious agents and or life stages of exotic (non-indigenous) species should not be discharged into surface waters. If not plumbed to sanitary lines (with proper chlorine disinfection) specialized systems to contain and treat contaminated waste will be constructed.
Polishing filters (particulate and/or charcoal) as well as water softeners can be provided to ensure the availability of conditioned water as necessary.
A dry area is to be provided for report writing, record keeping, and other water-protected activities. Dry areas are extremely hard to come by in aquatic facilities. This area should display procedures, emergency procedures, contact information, feeding schedules, water quality reports and any other important data.
Transportation of Aquatic Species:
North American Fishes:
Fishes captured within driving distance of University Park should be placed in 100L coolers equipped with aeration devices. The air pumps for these systems can be powered from 120-volt outlets or from a vehicle cigarette lighter. Water temperatures need to be monitored and adjusted by adding ice or heat.
Following is a description of the procedure that is followed for obtaining African fishes. A similar protocol should be followed if fishes from other sources as collected by the investigator. Fishes from Malawi are captured using SCUBA gear. All fishes caught at depths greater that 5 m are placed in underwater cages. These cages are moved toward the surface in 5 m intervals every 12 hours so that the fishes are decompressed slowly. Once on the surface, they are placed in plastic bags that are approximately 2/3 full of water. The remainder of the bag is filled with oxygen, sealed, and placed in insulated coolers. These coolers are then transported to Salima, Malawi, by either boat or motor vehicle to Stuart Grant's facility. Stuart Grant is the only licensed exporter of fishes in Malawi. The fishes are held in quarantine for a minimum of two weeks. After this period they are anesthetized, and packed as described above. They are then flown by either British Airways or KLM to Dulles International Airport. The necessary permits (see below) to import fishes into the United States through Dulles will be needed. The flight on which the fishes arrive is met at the airport, and checked through customs. They are immediately transported to University Park, where they are unpacked and placed in holding facilities.
A comprehensive Occupational Health Program is available at the University to support the full range of University research initiatives within the Aquatic Research Program. Specifically designed pre-placement, on-going, post-exposure, and termination medical surveillance examinations are readily available in the Occupational Health Clinic and can be provided for each protocol and for any identified potential chemical, physical, or biological health hazard exposure. Similarly, pre-placement and on-going surveillance examinations can be provided as needed to support hearing preservation, respiratory protection, laser safety, or other special area emphasis programs. Aquatic species do not customarily exhibit unusual susceptibilities to human disease. Individuals who notice cutaneous lesions on aquatic animals must handle the animals with care to avoid potential illness. If workers demonstrate skin lesions, they should report to the Occupational Health Clinic for timely medical evaluation and care.
Regulations Governing the Acquisition of Aquatic Species:
There are many regulations, which govern the shipping of aquatic organisms. Of particular interest are the Lacey Act, Endangered Species Act (endangered and threatened wildlife and plants), and Convention of International Trade in Endangered Species of Wild Flora and Fauna (CITES). Additionally, proper permits must be obtained and made available to USFWS personnel to collect these organisms. In certain cases (i.e. rare and or endangered species), permits are needed to hold these organisms, even if not collected by the investigator. With regard to specimens obtained outside the U.S., all import and export shipments must be declared. Obtaining clearance from an USFWS agent and filing a Form 3-177 is required at the time of entry or exit into or from the U. S. Within the U. S., the transportation of fishes across state lines without the proper permission and permits from the respective states and/or USFWS is in direct violation of the Lacey Act.
- Last Approved: Nov 12, 2012
- Last Revised: Dec 13, 2010