Purpose

In order to comply with the government regulations (USDA Animal Welfare Act and Animal Welfare Regulations, PHS Policy, and the Guide for the Care of Use of Laboratory Animals, 8th Edition), the UConn Health’s Institutional Animal Care and Use Committee (IACUC) has implemented a policy implementing a training form for individuals who receive training by the PI/laboratory staff.

Action

1. All individuals who perform research, testing, or training on laboratory animals must be trained on all procedures they will be expected to perform to comply with federal regulations.

2. All training must be documented in a written fashion.

3. If training is performed by the PI or senior laboratory staff members, the PI is responsible for maintaining training records for all procedures for which they train personnel to perform on laboratory animals.

4. All documentation of training must be submitted to the IACUC office when requested. IACUC office personnel must be allowed access to all training documentation records and be allowed to make copies.

Effective Dates:  June 2, 2022 through June 30, 2025

Purpose

Euthanasia is defined as the act of killing an animal by a method that induces rapid unconsciousness and death without pain or distress. In order to comply with federal regulations (USDA and PHS) and the Guide for the Care and Use of Laboratory Animals, the UConn Health’s Institutional Animal Care and Use Committee has established a policy on the use of CO₂ for euthanasia.

Action

1. Ideally, animals should be euthanized in their home cage.  If this is not possible (e.g., because an investigator is culling animals from multiple cages for euthanasia), the following will apply for transport of animals to the euthanasia station:
   1. The maximum cage density for euthanasia in a standard mouse cage will be 250 g/cage (approximately 8-10 mice)
   2. For cage densities greater than 125 g/cage in a standard mouse cage, animals must be euthanized within 30 minutes of being placed at these cage densities.
2. Compressed CO2 gas will be allowed to flow into an uncharged cage bottom or container.  Pre-filling of the chamber with CO2 is not allowed. As high concentrations of CO2 may be distressful to some species, gradual displacement of room air with the CO2 should allow for the anesthetic effect of CO2 to take place prior to asphyxiation which will result in the least amount of pain and/or distress to the animals.
3. For successive euthanasia procedures, the euthanasia chamber must be filled with room air before flooding with CO2 again.
4. To comply with the current AVMA Guidelines on Euthanasia, the flow rate into the euthanasia chamber must be at a rate of 30-70% of the total volume of the chamber per minute. To this end, a regulator must be used. The Euthanex® system is recommended by the IACUC; however, any commercial regulator system may be used as long as it meets the following criteria:
   1. The system must be AVMA compliant;
   2. The system must regulate a precise control of CO2 flow;
   3. The system must be calibrated specifically for CO2 gas; and
   4. The flow rate must be set by the factory and not be alterable by the user.
5. Death must be verified after euthanasia and prior to disposal. This typically should be done by visual examination (cessation of respiration and heartbeat), cervical dislocation, or thoracic trans-section.
6. CO2 euthanasia must be performed by individuals who are skilled in this method for the species in question and it must be performed in a professional and compassionate manner.
7. Unintended recovery of the animals after apparent death from CO2 is a documented occurrence. These incidents constitute serious non-compliance with PHS Policy and deviations from the provisions of the Guide.

References

1. Conlee, KM, Stephens, ML, Rowan, AN, and LA King. “Carbon Dioxide for Euthanasia: Concerns Regarding Pain and Distress, with Special Reference to Mice and Rats.” Laboratory Animals (2005) 30, 137-61.

2. Hawkins, P, Playle, L, Golledge, H, Leach, M, Banzett, R, Coenen, A, Coooper, J, Dannemann, P, Flecknell P, Kirkden, R, Niel, L, and M. Raj. Report of the Newcastle Consensus Meeting on Carbon Dioxide Euthanasia of Animals held on 2/27-28/06, University of Newcastle upon Tyne, UK, August 9, 2006.

3. PHS Policy on Humane Care and Use of Laboratory Animals Clarification Regarding Use of Carbon Dioxide for Euthanasia of Small Laboratory Animals. NIH Notice NOT-OD-02-062. July 17, 2002.

4. The Guide for the Care and Use of Laboratory Animals. National Research Council, 2011.

Effective Dates: June 2, 2022 through June 30, 2025

 

Purpose

In many cases, the use of drugs and chemicals in laboratory animals requires that a mixture be made. For example, ketamine and xylazine are frequently used by being made into a solution for an injection into animals. Or, alternately, drugs may need to be diluted in sterile saline prior to use.

Action

1. When making a mixture of drugs and/or chemicals in laboratory animals, documentation must be kept as to the identity of each individual drug/chemical used and their respective lot numbers and expiration dates.

2. If you are making drug mixtures (e.g., a working solution from stock) and plan to store the mixture for longer than the day it is made, mixtures must be made in a sterile container with a septum for re-use, such as sterile vials from Fisher Scientific or equivalent.  Alternately, a tube such as a vacutainer tube can be used; however, the stopper must be kept in and the drug mixture accessed by a needle using sterile technique (e.g., wipe septum with70% ethanol before obtaining mixture).  Sterile vacutainer tubes will be provided by Environmental Health & Safety should you choose to use that option.

3. If you are making a drug mixture and will use the entirety of it the day it is made, you can make your drug mixture in any sterile container (e.g., 15-ml Falcon Tube) as long as it is discarded that day.

4. Each mixture must be assigned an expiration date.  The expiration date will be the shortest expiration date of all drugs/chemicals used in the mixture, the expiration date of the sterile container used to make the solution, OR 1 month, whichever is soonest.

5. When storing expired controlled drugs in your laboratory, waiting for disposal with Environmental Health & Safety, the word “EXPIRED” must be on all containers.

 

Effective Dates:   December 7, 2023 through December 31, 2026

Purpose

The Public Health Service (PHS) policy on Humane Care and Use of Laboratory animals endorses US Government principles on use of Vertebrate Animals used in Testing, Research and Training, principle VII – “The living conditions of animals should be appropriate for their species and contribute to their health and comfort.” Based upon this, the Guide for the Care and Use of Laboratory Animals emphasizes “that animals should be housed with a goal of maximizing species-specific behavior and minimizing stress induced behavior”.

The primary aim of environmental enrichment is to enhance animal well-being by providing animals with sensory and motor stimulation, through structures and resources that facilitate the expression of species specific behavior and promote psychological well-being through physical exercise, manipulative activities, and cognitive challenges according to their species specific characteristics.

The environmental enrichment should be biologically relevant (i.e. hiding, gnawing, socializing, foraging, searching) so that it does not lose its enriching value over time. Environmental enrichment via physical manipulanda, structure, or substrate is preferred over nutritional enrichment.

All principal investigators working with laboratory animals are required to allow animal enrichment unless there is a scientific justification against the use of enrichment as approved by the Institutional Animal Care and Use Committee (IACUC). The justification must be included in an IACUC protocol submission and approved by the Attending Veterinarian (AV) and the IACUC.

Introduction of novel enrichment materials must be approved by the investigator and the AV before introduction.

Definitions

Physical manipulanda are non-nutritive objects that ca be altered by the animal or encourage fine motor movements. These include plastic/nylon chew toys – the preferred standard method of rodent enrichment at CCM.

Manipulanda are sanitized at least as frequently as the animal’s primary enclosure (cage).

Structure and substrate includes objects that allow an animal to isolate itself into different microenvironments, experience varied textures or materials. These include nesting materials, running wheels and hiding shelters.

Structure and substrate are replaced or sanitized at least as frequently as the animal’s primary enclosure (cage).

Nutritional enrichment may be provided in lieu of physical manipulanda, structure or substrate if necessary for research purposes.

Nutritional enrichment is accomplished by provision of novel foods or provision of small pieces of the normal diet in the bedding to stimulate foraging.

Procedure

Included in this are the Veterinarian Recommended Enrichment used currently at the CCM animal colony.

In addition, sometimes animals must be singly-housed due to various reasons such as incompatibility, pregnancy, surgery, medical isolation and other veterinary related issues.

Mice

• Social Housing: Animals are group housed whenever possible. Male mice may be housed singly due to their aggresive nature.
• Shelter: Nesting materials (e.g., Nestlets™); Igloos
• Other: Nylabones™

Rats

• Social Housing: Animals are group housed whenever possible. Male rats may be housed singly due to their aggressive nature.
• Shelter: Tubes (polycarbonate); Nesting materials may be desirable for dams nearing parturition and during lactation.
• Other: Nylabones™

Rabbits – most animals housed at CCM are male rabbits that are on surgical studies.

• Social Housing: All male rabbits are singly housed at CCM animal facility.
• Environmental Enrichment: Elevated shelves, toys (balls, bells, etc.)
• Dietary Items: Food items (e.g., alfalfa cubes/timothy hay/lettuce)
• Human Interaction: At least once a week, each rabbit becomes part of the human socialization program. The socialization consists of petting, brushing, to get the animals acquainted with human handling. Brushing reduces potential for hairballs.

Hamsters

• Social Contact: Group housed as much as possible; pair – or group-housed hamsters of same gender, except for pregnant females and incompatible animals.
• Nesting Material: Nestlet, tissue papers
• Other: Igloo, Jolly Ball™

Xenopus

• Being a prey species, Xenopus leavis are provided with shelter objects. The enrichment for X.leavis colony is the provision of PVC pipes. For the group housed X.leavis the number of enrichment (PVC) pipes depends on the density of the animals in the tank.
  • 1 to 5 frogs – 1 PVC pipe
  • 5 to 10 frogs – 2 PVC pipe

Fish

• The primary method of environmental enrichment is to be socially housed.  The use of artificial plants or other objects has been shown to have an effect on various aspects of fish metabolism.  If any artificial object is requested for the enrichment of fish requires the approval by the CCM Attending Veterinarian.

Implementation and Assessment

A successful well-being program depends on not only implementing but also on evaluating its effectiveness, and constant update to reflect our current knowledge. The principal investigators, research staff, veterinary staff and animal care technicians are involved at all times in decisions regarding psychological well-being and environmental enrichment.

References

1. Environmental strategies for laboratory animals. ILAR, Vol 46 (2), 2005.
2. “An Enriched Environment For The African Clawed Frog (Xenopus laevis)”: Lab Animal Magazine, May 1993, Vol. 22, Number 5, pp. 25-29.
3. Environmental enrichment program for hamsters. Institutional Animal Care and Use Committee, Cornell University, Ithaca, NY.
4. Guide for the Care and Use of Laboratory Animals – 8th edition. National Research Council, Washington DC. http://grants.nih.gov/grants/olaw/Guide-for-the-care-and-use-of-Laboratory-animals.pdf
5. Public Health Service Policy on Humane Care and Use of Laboratory Animals, OLAW, NIH. http://grants.nih.gov/grants/olaw/references/PHSpolicylabanimals.pdf

Effective Dates:      December 7, 2023 through December 31, 2026

Purpose

UConn Health’s Institutional Animal Care and Use Committee (IACUC) and the Attending Veterinarian are responsible for ensuring that all research animals are provided adequate veterinary care. This policy details requirements regarding tumor production to include implantable/inducible tumors, evaluating tumor growth, ascites produced by tumors, and endpoints and exemptions.

Action

Implantable/Inducible Tumors

1. Tumor implantation sites should be chosen to minimize damage to adjacent normal structures. The IACUC recommends implanting tumors on the dorsum or flank of an animal, as these areas will likely have minimal or no site-related morbidity.
2. Sites involving the face, limbs, or perineum should be avoided as there is little to no space for tumor growth and expansion.
3. Intramuscular implantation should be avoided as this is considered to be painful due to the distention of the muscle by the tumor.
4. Tumor implantation on the ventral surface of the body should also be avoided due to the risk of irritation to the tumor site in contact with the bedding and floor of the cage.

Evaluating Tumor Growth

1. Animals that are on a tumor production study must be monitored by the PI staff at least once a week during the time when the tumor is not yet detectable, to observe when tumor growth has begun.
2. After a visual or palpable tumor is evident, the animals must be monitored by the PI staff at least twice weekly. More frequent observations may be necessary based on tumor growth rate, study parameters, and general condition of the animals and will be detailed in the approved animal protocol.
3. The general physical condition of the animals is the most important factor in effectively following the progression of tumors in rodents.
4. When on the dorsum or flank of an adult rodent, tumors may be allowed to grow to mean diameters of 2.0cm in mice and 4.0cm in rats as long as the rodent remains otherwise healthy.
5. Evaluating tumors in central areas of the rodents (e.g., bone, lung) can be challenging. Tumor size will likely not be useful and, because of the sensitivity of areas, a small tumor may cause severe clinical signs. For tumor models studying central body tumors, clinical evaluation of the animals take priority over the measured size of the tumor. The expected clinical signs and the humane endpoints of those signs must be clearly described in the protocol. The evaluation of clinical signs in an animal with a tumor burden of this type should include consultation with the attending veterinarian.
6. Multiple tumors present other challenges. The concern of size for individual tumors is related to central necrosis, ulceration of skin overlying tumors, and abrasions. Multiple tumors that are individually smaller than the single tumor limit may not have the same negative sequelae as a single tumor. Multiple tumors may be allowed to grow up to 150% of the volume compared with the volume of a single tumor. Please note that the limitation on a single tumor will still be valid.
7. Tumor ulceration may not necessarily require euthanasia, but it does potentially require more frequent monitoring and treatment of the ulcerated tumor. The care for ulcerated tumors is based on both the size of the ulceration and the clinical judgment of the veterinarian.
   • Pinpoint (<=2mm) ulcerations at the site of tumor injection must be monitored at least 3 times per week for worsening of the ulceration site.
   • Ulcerations up to approximately 25% of the surface area of the tumor must be monitored at least 3 times per week and must be reported to the CCM veterinary staff for evaluation and potential treatment. If animals with ulcerated tumors of this size are not under veterinary care, these animals must be euthanized.
   • Ulcerations of greater than 25% of the surface of the tumor must be euthanized.

Species	Single tumor (max volume)	Multiple tumors (max volume)
Mouse	4.2cm3	6.3cm3
Rat	33.5cm3	50.25cm3

Ascites Produced by Tumors

1. In cases where tumors are expected to grow with an accumulation of ascites, rodents must be weighed prior to inoculation and subsequently be followed by weight measurements at regular intervals and this must be described in the approved protocol and based on the expected rate of fluid accumulation.
2. When the body weight exceeds 120% of the initial weight, the rodents must be euthanized or the animal tapped. Ascites pressure should be relieved before abdominal distention is great enough to cause discomfort or interfere with normal activity.
3. The abdominal tap should be performed by trained personnel using proper aseptic technique, with manual restraint or anesthesia, and by using the smallest needle possible that allows for good flow. This procedure must be described in the approved protocol.

Endpoints and Exemptions

1. Endpoints must be determined and specified in the experimental protocol.
2. If there is a strong scientific justification for maintaining tumors exceeding any of the guidelines described above, this must be detailed in the animal protocol and approved by the IACUC.
3. For studies that involve some degree of morbidity – Pain/Distress category D or E should be checked and the applicable sections filled out in the Animal Care and use Protocol.

References

1. The Guide for the Care and Use of Laboratory Animals, 8th Edition, 2012. National Research Council

2. IACUC Guideline on Rodent Tumor Production, University of Pennsylvania.

3. Ulman-Cullere MH and CJ Foltz. 1999. Body Condition Scoring: A Rapid and Accurate Method for Assessing Health Status in Mice. Lab Anim Sci 49(3): 319-323.

Effective Dates:      December 7, 2023 through December 31, 2026

Purpose

The Animal Welfare Act, Animal Welfare Regulations, and PHS Policy state that no animal work may be performed on protocols without IACUC approval which includes protocols that have expired. In addition, all approved protocols must completely describe the disposition of animals.

Action

1. All Principal Investigators (PI) who wish to transfer animals from an approved protocol to an approved protocol of another PI must have, in the original protocol, a statement that the disposition of animals includes the transfer of animals to another approved protocol.

2. The PI will need to inform the Center for Comparative Medicine (CCM) and the IACUC office of the request to transfer animals. The PI will need to give the following information: species and number of animals to be transferred, the name of the PI to whom the animals will be transferred, and the new IACUC protocol number. The IACUC office will confirm that the new protocol is approved.

3. This process may require that the original protocol be modified to include transfer of animals as a disposition method; this modification will require the approval of the Institutional Animal Care and Use Committee (IACUC) according to established procedures.

Effective Dates:  June 2, 2022 through June 30, 2025

 

Purpose

In order to ensure adequate review of animal care and use protocols, and to be compliant with both PHS Policy and recent clarification from the USDA, UConn Health’s IACUC has established a policy on assigning reviewers to new protocols.

Action

1. All protocols are submitted to the IACUC via the current electronic animal protocol and development system by the Principal Investigator.

2. The IACUC office will initially assign a primary (scientific) reviewer to the protocol. The primary reviewer assignment will be on a rotating basis with consideration of a reviewer’s particular area of expertise. In addition, the Attending Veterinarian (AV) and the Biological Safety Officer (BSO) and/or Chemical Safety Specialist (CSS) will also review all submitted protocols. If there are more than one AV, BSO and/or CSS on the IACUC, this assignment will be performed on a rotating basis.

3. The primary reviewers will remain for the life of the protocol; the initial primary reviewers will serve as the reviewers for all subsequent modifications to the protocol. If the initial primary reviewer is no longer a voting member of the IACUC when a future modification is received, the primary reviewer will be the Chair or Vice-Chair of the IACUC.  If the initial AV, BSO, or CSS reviewer not be a voting member of the IACUC when a future modification is received, these reviews will be performed by the current AV, BSO, or CSS on the IACUC committee.

Effective Dates: June 1, 2023 through June 30, 2026

This policy has been approved by a majority vote of the IACUC members.

Purpose

The Animal Welfare Act, Animal Welfare Regulations, and PHS Policy state that no animal work may be performed on protocols without IACUC approval which includes protocols that have expired. The Center for Comparative Medicine (CCM) has holding protocols on which animals may be temporarily transferred from a previously approved protocol (which has expired) so that the maintenance of these animals can be assured.

Action

1. All Principal Investigators (PI) are informed when any of their protocols expire. The PI is expected to comply with federal law that no experimental procedures may be performed on animals when the approved protocol expires. CCM is also informed when any protocol expires.

2. All animals under an expired protocol will automatically be transferred to the institutional holding protocol by CCM. While animals are being housed under this protocol, no experimental procedures- this includes breeding- are allowed. The holding protocol provides for routine husbandry and any veterinary care actions that may be required.

3. Once animals are transferred to the institutional holding protocol, there will be a card placed over the cage card which identifies the cage as being housed under the holding protocol. This should serve as a reminder that no experimental procedures may be performed on any animal in the cage.

4. The transfer of animals to the holding protocol will be allowed for three months ONLY. This will allow the PI time to write a new animal care and use protocol and have it approved by the Institutional Animal Care and Use Committee.

5. Animals transferred to the institutional holding protocol will be euthanized, or otherwise appropriately disposed of, once the three months expiration date is reached unless the PI of the original expired protocol requests a one-time extension of an additional three months.

Effective Dates:  June 2, 2022 through June 30, 2025

Purpose

In order to ensure a safe environment for both employees of the Health Center, and the laboratory animals, the Institutional Animal Care and Use Committee (IACUC) has implemented a policy regarding the transport of animals in the Health Center.

Action

1. Animals must be transferred into clean caging prior to removal from the animal facility.

2. Animals must be transported in closed caging (either microisolator lids or soft bonnet type covers) and covered with an opaque material (e.g., sheet, towel, etc.).

3. If more than 1 cage of animals is being transported, transport must be on a cart because of the possibility of dropping the cages.

4. Animals may not be transported on a common-use elevator. CCM elevators only are to be used to transport animals between floors.

5. Animal caging (with or without animals) may NOT be placed, unsupervised, in the hallways. All caging must be kept in the laboratory areas.

6. Animals are to be transported in commercial or CCM-operated vehicles ONLY unless a modification of an approved protocol to transport animals in personal cars has been approved by the IACUC.  Please refer to the IACUC Policy on Mouse/Rat Transportation Outside UConn Health for transport of animals outside the institution.

 

Effective Dates: June 6, 2024 thru June 30, 2027

Purpose

In order to comply with government regulations and to simplify the approval process, the IACUC has established a policy on the ability to utilize the designated member review (DMR) for approval of new protocols.

Action

IF THE ORIGINAL FULL COMMITTEE REVIEW RESULT IS “MODIFICATIONS REQUIRED”, THEN:

1. The changes requested to the protocol are sent to the Principal Investigator (PI) of the protocol through the InfoEd eRA portal Animal Use / IACUC module.

2. When the PI has responded to the request for changes, the PI’s changes are sent to the reviewers (Primary, veterinarian, research safety) of the protocol via InfoEd with a request to review the changes by a specified date (generally one week).

3. All reviewers must agree to the changes made. If there is no consensus to the changes, the protocol will go back to the full committee.

4. When all three (3) reviewers have indicated approval of the change(s) in InfoEd, the IACUC office will approve the protocol.

Effective Dates:  June 1, 2023 through June 30, 2026