Standard Operating Procedures

Zebrafish Health Monitoring Program

Last updated: March 5, 2012

I. PURPOSE

Health monitoring and routine sentinel surveillance of zebrafish for infectious disease is an important aspect of fish husbandry at The University of Montana (UM).

II. RESPONSIBILITY

It is the responsibility of the Laboratory Animal Resources (LAR) staff to understand the health monitoring program for zebrafish, to monitor mortality and disease events, to collect sample fish, and to submit fish samples for analysis and diagnosis.

III. PROCEDURE

• Fish Tanks
  • Zebrafish in the LAR facility are held in groups of 7-20 fishes in small, individual fish tanks on an Aquatics Habitat Re-circulating Fish Rack System (ZF0601)
  • Water tests for fish tanks
    • Tank has digital monitoring system.  Reads pH, temperature and dissolved oxygen
    • Weekly, test for ammonia, nitrite and salinity in all tanks
    • Monthly, test for alkalinity, iron, dissolved oxygen and hardness in all tanks
  • Review water monitoring data on a quarterly basis to identify any unusual trends in the data.
• Disease Event
  • Notify the Attending Veterinarian (AV) of any mortality or disease events in the zebrafish population.
  • Common signs indicating disease in zebrafish are body wasting and spinal deformity
  • Review the recent water monitoring history for any anomalies.
  • Collect any fish found dead and fix in formalin. Contact AV for evaluation of fish before shipping.
  • Collect 1-2 live fish that are demonstrating the clinical signs of the disease in question.
    • Euthanize fish with MS222, 200-500 mg/L. Wait 10 min after last opercular movement before putting in formalin.
    • Fix fish in formalin.
    • Package fish appropriately for mailing, include a Veterinary Diagnostic request form in the outer package, and mail to the Oregon State University Vegerinary Diagnostic Laboratory.
• Sentinel Surveillance
  • Euthanize 1 fish per year, rotating which tank it comes from each year (July).
    • Euthanize in MS222, 200-500 mg/L
    • Take gill and fin clips of the euthanized fish for wet mounts.
    • Fix the euthanized fish in formalin and send to a fish pathologist for evaluation.
    • Package fish appropriately for mailing, include a Veterinary Diagnostic request form in the outer package, and mail to the Oregon State University Veterinary Diagnostic Laboratory.
  • Anesthetize 1 fish per tank every year (January) on an alternate schedule from #1 above so that health-checks are conducted every 6 months.
    • Anesthetize in MS222 at 75 mg/L
    • Take gill and fin clips of the anesthetized fish for wet mounts.
    • Allow fish to recover from anesthesia in a container of water from home tank and return fish to home tank.
• Reporting Test results
  • The Oregon State University Veterinary Diagnostic Laboratory will report results to the UM Attending Veterinarian.
  • The AV will email a copy of the report to LAR staff members and to the PIs working with zebrafish.
  • Test results will be maintained by the AV for a minimum of three years.