SOP: # 302 Effective Date: 2/3/2022 AQUATIC ANIMAL FACILITY MAINTENANCE Supersedes Document Dated: 3/18/2020 Page 1 of 13 Approved 2/3/2022 1. PURPOSE: This SOP has been created to standardize the minimum requirements in maintaining an aquatic animal facility. More frequent intervals for maintenance of the below items are encouraged. This SOP excludes outdoor aquatic animal housing (i.e. ponds, mesocosms). The care and maintenance for these housing types should be described in the approved protocol for animal use. Exceptions to this SOP must be approved by IACUC in the animal use protocol. 2. PROCEDURES 2.1 Housing and Husbandry 2.1.1 Housing Density (a) The Guide for the Care and Use of Laboratory Animals recommends a housing density of 10 adult zebrafish per liter, and 2 liters per adult Xenopus frog. Younger animals may be housed at higher densities. For zebrafish, 20-200 eggs or embryos/100ml, 20 larva 3-30dpf/400ml, 5-10 fish >30dpf/L. (b) Housing density should allow for adequate space between animals and promote well-being. (c) Each tank must be individually identified. 2.1.2 Tank Sanitization and Frequency (a) Tanks should be checked for algae accumulation daily and cleaned as necessary. Algae must not inhibit viewing of the animals. (b) Reusable enclosures must be sanitized between use to prevent the spread of pathogenic micro-organisms, animal wastes and allergens. (c) Tanks must be removed and sanitized at regular intervals, not to exceed 60 days for zebrafish. For other species, sanitization of tanks should occur at an appropriate interval for the species and research performed. As tanks or rows of tanks/shelves are sanitized this must be recorded. Care must be taken to thoroughly rinse any chemical residue prior to use. (d) Tanks should be suctioned/spot cleaned as needed to remove debris.
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SOP: # 302 Effective Date: 2/3/2022
AQUATIC ANIMAL FACILITY MAINTENANCE
Supersedes Document Dated: 3/18/2020
Page 1 of 13
Approved 2/3/2022
1. PURPOSE:
This SOP has been created to standardize the minimum requirements in maintaining an
aquatic animal facility. More frequent intervals for maintenance of the below items are
encouraged.
This SOP excludes outdoor aquatic animal housing (i.e. ponds, mesocosms). The care
and maintenance for these housing types should be described in the approved protocol for
animal use.
Exceptions to this SOP must be approved by IACUC in the animal use protocol.
2. PROCEDURES
2.1 Housing and Husbandry
2.1.1 Housing Density
(a) The Guide for the Care and Use of Laboratory Animals
recommends a housing density of 10 adult zebrafish per liter, and 2
liters per adult Xenopus frog. Younger animals may be housed at
higher densities. For zebrafish, 20-200 eggs or embryos/100ml, 20
larva 3-30dpf/400ml, 5-10 fish >30dpf/L.
(b) Housing density should allow for adequate space between animals
and promote well-being.
(c) Each tank must be individually identified.
2.1.2 Tank Sanitization and Frequency
(a) Tanks should be checked for algae accumulation daily and cleaned
as necessary. Algae must not inhibit viewing of the animals.
(b) Reusable enclosures must be sanitized between use to prevent the
spread of pathogenic micro-organisms, animal wastes and
allergens.
(c) Tanks must be removed and sanitized at regular intervals, not to
exceed 60 days for zebrafish. For other species, sanitization of
tanks should occur at an appropriate interval for the species and
research performed. As tanks or rows of tanks/shelves are sanitized
this must be recorded. Care must be taken to thoroughly rinse any
chemical residue prior to use.
(d) Tanks should be suctioned/spot cleaned as needed to remove
debris.
SOP: # 302 Effective Date: 2/3/2022
AQUATIC ANIMAL FACILITY MAINTENANCE
Supersedes Document Dated: 3/18/2020
Page 2 of 13
Approved 2/3/2022
(e) Recirculating and flow-through systems should have partial water
changes daily to semi-weekly.
(f) Static tanks with animals over 30 dpf (fish) or stage 30
(amphibians) should have weekly water changes of at least 75%.
Animals less than this age should have water changes as
appropriate.
(g) Lids must be wiped off at least once weekly to remove excess food
and debris.
2.1.3 System Maintenance
(a) System water level must be checked daily and refilled as needed.
(b) If a UV light is used, the indicator light must be checked daily.
(c) Baffles should be checked weekly and cleaned as necessary.
(d) Mechanical filters should be changed or cleaned as often as
necessary or at least monthly.
(e) Carbon filters should be changed monthly.
2.1.4 Feeding
(a) Feeding frequency, amount, and type is to be determined at the
discretion of the investigator. It must be recorded at each instance
and animals should be fed on a regular schedule.
(b) Prepared foods must be discarded either 6 months after being
received or opened or at the manufacturer’s expiration or best-by
date. All food containers must be sealed and labeled with the date
of production, date of receipt, and/or date opened, as well as the
expiration date. Freezing the food prior to the expiration date will
extend the use-by date by 6 months. The date frozen must be
labeled in addition to the updated expiration date.
(c) Live foods should be cultured on the premises and derived from
pathogen-free environments.
2.1.5 Enrichment - The support of enrichment in aquatic animal enclosures is
variable in the literature. If used, it should be safe for the animals.
SOP: # 302 Effective Date: 2/3/2022
AQUATIC ANIMAL FACILITY MAINTENANCE
Supersedes Document Dated: 3/18/2020
Page 3 of 13
Approved 2/3/2022
(a) Zebrafish- current literature suggests that enrichment is beneficial
to reduce aggression in low density housing scenarios. It also
suggests plastic plants may enhance fecundity in breeding pairs.
Classical music may also benefit zebrafish by reducing stress.
(b) Wild-caught animals should be provided plastic plants or a device
to hide in to reduce stress associated with captivity. Eggs hatched
in captivity are not considered wild-caught for this purpose.
(c) Frogs must be provided a shelter to hide in to reduce stress.
2.2 Water Quality
2.2.1 Monitoring
(a) Water quality monitoring must be performed for each system for
recirculating and flow-through systems, or for each tank if static.
For static tanks, if they are on the same water change schedule, one
tank per shelf may be tested to represent the whole shelf. In these
cases, the tank with the highest density should be tested.
(b) Water temperature must be monitored daily.
(c) Dissolved oxygen must be monitored daily for fully aquatic
species or life stages.
(d) Water quality parameters should be based on species, type of
aquatic system, animal density, and whether the system is
established versus new. New systems require more frequent testing
than those that are established.
(e) For static systems, if complete water change occurs at least
weekly, no testing of pH, ammonia, nitrate, or nitrite is needed.
Otherwise weekly testing is required.
(f) The following parameters must be tested at least monthly in
established recirculating and flow-through systems, and at least
weekly in static systems.
(i) Ammonia (NH3) should remain at 0mg/L
(1) For production fish (aquaculture), the following
recommendations apply.
SOP: # 302 Effective Date: 2/3/2022
AQUATIC ANIMAL FACILITY MAINTENANCE
Supersedes Document Dated: 3/18/2020
Page 4 of 13
Approved 2/3/2022
a. Salmonids – Unionized Ammonia less than
0.0125 mg/L
b. Cool-water fish – Total Ammonia Nitrogen
less than 1.0 mg/L
c. Warm-water fish – total Ammonia Nitrogen
less than 3.0 mg/L
(ii) Nitrite should remain under 0.1mg/L
(iii) Nitrate should remain under 200mg/L
(iv) pH
(v) Chlorine (for systems not supplied by RO water or well
water) should remain at 0mg/L
(g) It is recommended to monitor other parameters such as hardness,
alkalinity, conductivity, and total gas pressure monthly or more
often as appropriate for the species.
2.3 Health Monitoring
2.3.1 Daily Health Observations
(a) Daily health observations of each tank must be performed to
evaluate the health of the animals.
(b) Health should be evaluated by using body condition scoring
(Appendix B) and the Health Evaluation Chart (Appendix A)
below.
(c) Dead and euthanized animals must be recorded on a mortality log
daily for animal of life stages large enough to individually count,
for example zebrafish 30dpf and older. Some mortality is
anticipated in large colonies and with developing fry. If within
expected ranges, mortality data can be provided to LAP in a
monthly summary.
2.3.2 Monthly Health Observations
(a) Within the first week of each month, health, mortality, and census