Washington University in St. Louis School of Medicine

Zebrafish Facilities

WUSTL-SOP-002: Zebrafish Quarantine

Bringing new lines to Washington University in St. Louis School of Medicine Zebrafish Facilities must to be carried out with caution to minimize danger of “importing” diseases. The following steps should be taken:

  1. An investigator who wishes to import into Washington University Zebrafish Facilities a zebrafish strain of certain genotype should first obtain an approved protocol at IACUC for maintaining this line at Washington University in St. Louis School of Medicine.
  1. All lines (arriving as adult fish or larvae) are introduced into our Quarantine system, currently located in room 3910 South Building. Under no circumstances should new arrivals be placed in any of the main Fish Facilities. Please notify Facility Staff of any and all incoming adult fish, larvae, or embryos.
  1. All lines (arriving as adult fish or larvae) should be assigned a Washington University Quarantine (Q) number. Facility staff will enter lines into the Quarantine stock list, once provided the appropriate information.
  1. All lines (arriving as adult fish or larvae) should be marked with a proper label containing Q number, birth date, genotype and responsible investigator.
  1. In case of imported larvae: at 5 days post fertilization, place them in a 3 Liter tank, with 800m screen. Place the tank in the Q-system next to other babies, if present. If no baby tanks are currently present in the system, mark the tank below with tape “babies” to alert the feeders and ensure proper feeding and care
  1. In case of imported adult or juvenile fish: Please alert facility staff of all incoming fish lines. Do not place adults into the system. Adult alien fish should be placed in a tank containing a false bottom on the shelves next to the door; this is the responsibility of facility staff. At this point the fish should be monitored for Caplilaria worms and preventively treated with Fluke tabs (see separate SOP#). 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 (Q number, birth date and genotype, responsible investigator), and appropriate feeding label.
  1. Transfer of genotypes into the main Fish Facility. When the larvae grow into adults, or imported adults need to be outcrossed in the Q-system using wild-type AB* fish (or another strain when appropriate), their progeny must be bleached by Facility Staff (use Bleaching Zebrafish Embryo SOP) and then transferred the Nursery.
  1. The owners of imported lines are responsible for notifying Facility Staff to remove them (euthanizing) when the progeny of desired phenotypes has been transferred and established in the main Fish Facilities.

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.
  3. Absolutely nothing can be transferred from the Q-system to the main Fish Facilities, unless autoclaved, or as bleached embryos.