Washington University in St. Louis School of Medicine Zebrafish Facilities WUSTL-SOP-008: Use of Fluke Tabs for Prophylactic treatment of Capillariasis in Quarantined Adult Zebrafish

From WUSTL-SOP-002: psuedocapillaria tormentosa In case of imported adult or juvenile fish: Do not place adults into the system. Adult alien fish should be placed in a tank containing a false-bottom (e.g. mating cage) on the shelves next to the door, this is the responsibility of the researcher who imports the fish. At this point the fish should be monitored for Capillaria worms (by examining the waste that collects in the bottom of the tank) and preventively treated with Fluke tabs (see below). As the fish are treated for worms, they should be also monitored for development of lesions on the body or any other clinical manifestations of pathogens. After the treatment and monitoring for worms and Tuberculosis fish can be transferred to the system, in appropriate tank (1.1L up to 11 fish or 3L or 8L if more than 15 adults), appropriate label (WU number, birth date and genotype, responsible investigator), and appropriate feeding label.

Use of Fluke tabs to prophylactically treat adult or juvenile zebrafish before they are allowed on to the recirculating system:

1. add the fluke tabs aliquot (13.2mg) to a 50mL conical tube with system water and dissolve as thoroughly as possible using a vortexing mixer; this will make 1L of treatment water. 2. add the treatment water to the mating cage with a false-bottom insert. 3. Place the fish to be treated into the mating cage (maximum 4fish/mating cage) 4. At 24-hrs, and after disposing of 1/3 of the water from the first treatment, add another 13.2mg of Fluke Tabs to the mating cage, and top up to 1L with fresh fish water (bringing the concentration of drug from 13.2mg/L to 22mg/L). 5. At 48 hours, repeat step #4 (bringing the final concentration of drug to 27.86mg/L) 6. If, at 72-hours, no evidence of capillaria (adult worms or their eggs) have been found, the fish may be introduced to the recirculating Quarantine System by removing them from the Fluke Tabs treatment water and placing them in an appropriate tank (1.1L up to 11 fish or 3L or 8L if more than 15 adults), with an appropriate label (WU number, birth date and genotype, responsible investigator). If continued evidence of capillaria is found, start the treatment over from step 1 for another full round of treatment.

Adult capilllaria worm Capillaria egg

Important: When working in Q-system:

1. Always wear gloves and discard them after use. 2. Attend the Q-system only after any work in the main Fish Facility is completed. If you need to return to the main Fish Facility after working in the Q-system, wash your hands and elbows thoroughly. 75% Ethanol (ETOH) is available for this purpose as well. 3. Absolutely nothing can be transferred from the Q-system to the main Fish Facilities, unless autoclaved, or as bleached embryos. Washington University in St. Louis School of Medicine Zebrafish Facilities WUSTL-SOP-008: Use of Fluke Tabs for Prophylactic treatment of Capillariasis in Quarantined Adult Zebrafish