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UNIVERSITY OF SOUTHERN MAINE
Office of Research Integrity & Outreach
Procedure #:
Date Adopted:
Last Updated:
Prepared By:
Reviewed By:
Procedure Title:
04-003
7/12/2012
Sarah Gilliam, Animal Facilities Manager
Jeffrey Walker, Associate Professor, Biological Sciences
USM IACUC Committee
Fish Care & Husbandry
1.0 Purpose: To provide a comfortable, clean & healthy environment for all fish
according to guidelines and regulations.
2.0 Responsibility: It is the responsibility of the investigator, technician, or student
performing this task to follow the procedures established in this document.
3.0 Applicable Documents:
3.1 Guide for the Care and Use of Laboratory Animals
3.2 SOP 02-001: Animal Space Provision
3.3 SOP 03-001: Daily Observation of Laboratory Animals
3.4 SOP 03-002: Animal Carcass Disposal
3.5 SOP 03-005: Animal Identification
3.6 Equipment operating manual(s)
3.7 Zoonotic Diseases List
4.0 Safety Considerations:
4.1 Animal mishandling can result in injury or illness to the animal.
4.2 Read all labels on disinfectants carefully to avoid any harmful exposure to the
technician or animal.
4.3 Refer to the zoonotic diseases list posted within the facility.
5.0 Housing & Environment:
5.1 Appropriate caging/housing:
5.1.1 Aquatic Habitats Zebrafish Rack, tanks ranging from 1-3 gallons.
5.1.2 140g Instant Ocean
5.1.3 16ml sodium bicarbonate solution
5.1.4 40L distilled water
5.1.5 Follow instructions in the manual for the rack for preparation &
monitoring of water quality.
5.2 Water temperature should be maintained at 18-20oC
5.3 Lighting within the room should be on a 16 hour light, 8 hour dark cycle.
5.4 Tank Maintenance-Tanks should be cleaned weekly with via vacuuming
with a siphon. Tanks should also be scrubbed with an acceptable pad such as
scotch brite pads on an as needed basis to remove algae.
5.4.1 Siphon-Attach tip of siphon to its hose. The tip is the only part of
the siphon that should be immersed in the tank water. Start siphon
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5.4.2
5.4.3
by submerging the tip and running water into the hose until it is
filled. Turn the valve on the hose so that the water is ‘trapped’ in
the hose. Put the tip back into the water and turn the valve,
creating suction. Vacuum all waste off of the bottom of the tank,
taking care to avoid the fish.
5.4.1.1 After completing the siphoning of all tanks, clean/disinfect
the tip of the siphon by soaking in a bleach solution (20
parts water to 1 part bleach). Follow that with a rinse in
sodium thiosulfate solution for removal of chlorine.
Scrub-Scrubbing the sides of the tank(s) will remove algae. If
visibility into the tank is obscured, then it is past time to scrub the
tank. Tanks should be checked daily to determine if algae removal
is necessary.
Water testing should be done on a regular basis to ensure water
quality and safety of the fish.
6.0 Feeding:
6.1 Feeding is done once per day.
6.2 Acceptable foods:
6.2.1 Choice of food should be done in consideration to the species of
fish, size, and age of the animal(s).
6.2.2 Fry-250-450 microns of newly hatched brine shrimp or Larva Z
Plus (commercially available diet).
6.2.3 Juveniles/Adults-frozen squid (slice thinly while frozen), Silver
Cup Trout Chow #1 or #2 (2,590 microns-1.38 mm), freeze-dried
blood worms, freeze-dried brine shrimp, or commercially available
‘flake’ diet.
6.2.4 Enough food should be given that it is all consumed within 5-10
minutes. The amount should also be of a quantity that no food
remains on the bottom of the tank after 30 minutes.
6.3 Food Storage & Handling:
6.3.1 All incoming foods need to be dated with the received date & with
the date opened.
6.3.2 Food can be stored for up to one year in the freezer, unless
otherwise specified by the manufacturer.
6.3.3 Dry foods are kept in a -20oC freezer and partitioned from there.
6.3.4 Food partitions are then kept in a 4oC cooler, in a secondary tub.
7.0 Transport:
7.1 All animal orders or transfers need to be coordinated with the Animal
Facilities Manager prior to arrival.
7.2 Via Car/Vehicle: Fish should be placed in plastic bottles or bags filled at ½
volume with the animal’s source water (water from the tank or enclosure from
which the fish came). Close container with lid or knot to prevent water
leakage, escape, & harm to the animal.
7.2.1 Take care not to over crowd containers, small species such as
sticklebacks, zebrafish, & medakas should be no more than 6-12 per
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2L container. Larger species should have spacing adjusted
accordingly.
7.2.2 Containers should be placed into a cooler (packed with ice or ice
packs), and when possible an air stone attached to a battery operated
air pump should be placed into the container to provide extra
oxygenation of the water during transport.
7.3 Via Plane or Express Parcel Service: Follow the same steps as in 7.2 above.
Pack containers with freezer packs in a cooler and seal the cooler closed with
tape. Cooler should be shipped with overnight delivery.
8.0 Introducing the Fish to New Housing: Fish should be acclimated to their new
environment by exchanging half of the water in their transport containers with water
from their new tank.
8.1 A serious of 3 changes, with 30 minutes between each change will enable a
slow acclimation to their new tank. Once 3 changes have occurred, with a 30
minute wait after the last change, the animals can then be transferred into the
laboratory tank.
9.0 Sick & Dead Animals: Animals should be checked daily.
9.1 If a sick fish is found, then the Principal Investigator should be notified
immediately and a determination can be made whether to treat the animal or
euthanize it.
9.2 If a dead fish is discovered, make a note on the tank, change the quantity of
animals on the tank, and record on the daily census sheet.
9.2.1 The dead fish should be removed from the tank, and either placed in a
previously determined spot (such as a small bucket in a freezer or preserved in
a sample jar/vial) as instructed by the Principal Investigator.
9.2.2 Once the PI is done collecting tissues, etc, then the carcass should be
placed in the Animal Facility carcass freezer for disposal.
10.0 Fish Euthanasia: This should only be performed by trained individuals &
performed in accordance to the AVMA Guidelines on Euthanasia.
10.1 Combine 7.5ml of MS222 (Tricaine) into 100 ml of tank water in bucket or
container. Increase volume of the Tricaine/tank water solution as necessary
for the size of the fish.
10.1.1 The steps for making the MS222 stock solution can be found in The
Zebrafish Book, vol. 4.
10.2 Place the fish into the Tricaine/tank water solution and leave there until at
least 10 minutes past opercular movement has ceased.
10.3 If fish is to be preserved for tissues or further study, then proceed with
fixation. If fish is to be disposed of, then it can be placed into a designated
container by the PI or into the Animal Facility carcass freezer for disposal.
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