Purpose

Many Tg/GT animals are being constructed that require monitoring by the IACUC to comply with various federal regulations. When reviewing protocols, the IACUC has the responsibility to ensure the necessity for the use of live vertebrate animals and to ensure the appropriate use of species and strains for the proposed work.

Action

1. Any Tg/GT animal to be constructed and/or used at UConn Health must be named and described in an approved animal care and use protocol. This includes animals destined for use at either the UCHC or an outside institution.

2. All Tg/GT will be assigned a Genetically Modified Organism (GMO) number by the BSO or IACUC coordinator.  This GMO number must be reflected on the cage cards housing Tg/GT animals.

3. It is the responsibility of the PI to inform the IACUC if there are any phenotpyic pain or distress associated with the line, including unexpected lethality.  If this is not known at the time of initial use, it must be reported to the IACUC immediately upon discovery.

4.  Protocols must clearly indicate the recombinant lines to be used in that protocol.

 

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

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

Purpose

In order to ensure appropriate use of Tribromoethanol (Avertin), UConn Health’s Institutional Animal Care and Use Committee (IACUC) has implemented a policy on the use of this drug. The IACUC encourages the use of pharmaceutical grade compounds such as ketamine/xylazine and isoflurane rather than Tribromoethanol.

Background

Tribromoethanol (Avertin) is an anesthetic that provides rapid induction and recovery for single use, short duration (approximately 15-20 minutes) surgical procedures in rodents. Tribromoethanol has been commonly used in the production of transgenic animals to facilitate procedures such as embryo transfer, vasectomy, or distal tail snip for DNA analysis. Tribromoethanol is not commercially available and investigators who wish to use it as an anesthetic must make their own solutions. Improper preparation, storage, or use of Tribromoethanol can result in high mortality losses. In particular, Tribromoethanol degrades in the presence of heat and light, producing toxic by-products that are potent gastrointestinal irritants. Adverse effects are common in mice following any second exposure to Tribromoethanol regardless of the dosing interval; therefore, this anesthetic is approved for one administration only as a survival anesthetic in mice.

Any proposed deviation from these guidelines must be fully explained and justified to the IACUC in the research proposal.

Action

Preparation

Tribromoethanol must be prepared as follows:

• Dissolve 2.5 grams of 2,2,2-tribromoethanol in 5 ml of tert-amyl alcohol.  This requires heating to approximately 40°C and stirring vigorously.
• Add sterile PBS, stirring continuously, up to a final volume of 200 ml
• Filter sterilize through a 0.5 micron filter
• Aliquot the final solution into sterile light protected containers.  Containers must be labeled:
  • Tribromoethanol
  • 12.5 mg solution
  • Date of preparation
  • Date of expiration (two weeks from reconstitution)
  • Initials of person who prepared the solution

Storage

Tribromoethanol must be stored at 2-8°C in light protected containers.

Use

Tribromoethanol may be approved for use after scientific and/or medical justification has been provided has been provided in the animal care and use protocol and after IACUC review for a single, survival administration to adult mice. If a second administration of tribromoethanol is given, the animals must be euthanized prior to awakening from the anesthetic. The anesthetic should be administered at a dose of 250 mg/kg given IP.

Dispose of any solution that is past the two-week expiration date, has crystals, or has changed from a clear solution to a yellow solution.

References

1. Papaioannou, VE and Gox, JG. Efficacy of Tribromoethanol Anesthesia in Mice. Laboratory Animal Science, 1993. April, 43(2): 189-192.

2. Zeller, WM; Burki, G; and Panoussis, B. Adverse Effects of Tribromoethanol as Used in the Production of Transgenic Mice. Laboratory Animal Science, 1998. October, 32(4): 407-413.

3. Kohn, DF; Wixson, SK; White, WJ; and Benson, GJ. Anesthesia and Analgesia in Laboratory Animals, 1997.

4. Efficacy and Safety of Stored and Newly Prepared Tribromoethanol in ICR Mice. Contem Top Lab Animal Sci, 2005. 44(1): 17-22.

5. An Evaluation of Preparation Methods and Storage Conditions of Tribromoethanol. Contem Top Lab Animal Sci, 2005. 44(1): 11-16.

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

Purpose

There are many circumstances that involve partnerships between collaborating institutions or relationships between institutional animal care programs. OLAW and USDA-APHIS agree that review of a research project or evaluation of a program or facility by more than one recognized IACUC is not a federal requirement.

It is imperative that institutions define their respective responsibilities. PHS Policy requires that all awardees and performance sites hold an approved Animal Welfare Assurance. OLAW negotiates Inter-institutional Agreement Assurances of Compliance when an awardee institution without an animal care and use program or IACUC will rely on the program of an Assured Institution. Assured institutions also have the option to amend their Assurance to cover non-assured performance sites, which effectively subjugates the performance site to the Assured institution and makes the Assured institution responsible for the performance site.

If both institutions have full PHS assurances, they may exercise discretion in determining which IACUC reviews research protocols and under which institutional program the research will be performed. It is recommended that if an IACUC defers protocol review to another IACUC, then documentation of the review should be maintained by both committees. Similarly, an IACUC would want to know about any significant questions or issues raised during a semi-annual program inspection by another IACUC of a facility housing a research activity for which that IACUC bears some responsibility or exposure. This policy defines when UConn Health will accept another institution’s IACUC approval and/or require submission to the UConn Health’s IACUC.

Action

1. Work performed at UConn Health with the funding administered through another institution:

• The UConn Health’s IACUC will review and approve all animal care and use activities performed at this institution. The work must have a UConn Health faculty member sponsor and the Principal Investigator must complete the UConn Health’s animal care and use protocol form. All personnel involved in the study performed at this institution must comply with all IACUC policies and procedures.

2. Work performed at another institution with the funding administered through UConn Health:

• UConn Health’s IACUC will accept the review and approval of the institution performing the work if the following are met:
  • UConn Health’s IACUC receives a copy of the protocol approval letter from the institution.
  • UConn Health’s IACUC receives a copy of the IACUC approved protocol from the institution.
  • UConn Health’s IACUC receives a copy of the latest semi-annual inspection report to the IO.
  • Additional documentation may be requested at the discretion of the IACUC chair.
• If any of the above requirements are not met, then the PI will be required to submit a protocol to the UConn Health’s IACUC for review.

3. Work performed at UConn Health and another institution with the funding administered through UConn Health:

• This will be handled for each institution as described in 1 and 2 above.

4. Work performed at UConn Health under an agreement which may be for pay or collaboration for an institution or company:

• If the institution or company has an IACUC, the UConn Health’s IACUC will accept the review and approval of the institution performing the work if the following are met:
  • UConn Health’s IACUC receives a copy of the protocol approval letter from the institution.
  • UConn Health’s IACUC receives a copy of the IACUC approved protocol from the institution.
• If the institution or company does not have an IACUC, the work will be performed as described in 1 above.

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

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

Purpose

In order to comply with government regulations (PHS and USDA), the Association for the Assessment and Accreditation of Laboratory Animal Care International (AAALAC), and the Guide for the Care and Use of Laboratory Animals, the University of Connecticut’s Institutional Animal Care and Use Committee (IACUC) has implemented a policy regarding semi-annual program review and facility inspections.

Action

1. Animal study area is defined as “any building, room, area, enclosure, or other containment outside of a core facility of centrally designated or managed area in which animals are housed for more than 12 hours if it houses a USDA regulated species and more than 24 hours for other species.”

2. The Office of Laboratory Animal Welfare (OLAW), NIH, has issued a document which states “when considering IACUC responsibilities for semi-annual review, it is important to keep in mind that the institution, acting through the IACUC, is responsible for all animal-related activities of the institution, regardless of where animals are maintained or the duration of their stay.”

3. In May and November of any calendar year, the IACUC will conduct a facility inspection to include all CCM areas including, but not limited to: animal holding rooms (including any satellite facilities), animal study areas, cage wash areas, food and bedding storage areas, and any other area deemed appropriate. In addition, all laboratories in which animals are listed as being used will be inspected along with laboratories where animals are held for more than 12 (USDA-regulated species) or 24 hours (all other species).

4. Once the inspections are completed, the IACUC will hold a special meeting to review the animal care and use program and the results of the facility and laboratory inspections. A report will be generated by the IACUC coordinator which will be signed by a majority of a quorum of the IACUC and will include any minority views. This report will then be submitted to the institutional official.

Effective Dates:   December 1, 2022 through December 31, 2025

Purpose

Varieties of acceptable and recognized methods are available for individually identifying research animals. These include, but are not limited to, ear tags/notches, tattoos, sutured beads, subcutaneous transponders, colored stains, dyes, and freeze brands. The Guide for the Care and Use of Laboratory Animals states “As a method of identification of small rodents, toe-clipping should be used only when no other individual identification is feasible.  It may be the preferred method for neonatal mice up to 7 days of age as it appears to have few adverse effects on behavior and well being at this age, especially if toe clipping and genotyping can be combined.  Under all circumstances, aseptic practices should be followed.  Use of anesthesia or analgesia should be commensurate with the age of the animals.”

Toe clipping is the removal of the last phalangeal (toe) bone of a digit from one or more limbs. It is usually done for the purpose of identification and genotyping of animals. The Food and Drug Administration also discourages the use of toe clipping as a means of animal identification. The UCHC Institutional Animal Care and Use Committee (IACUC) typically does not approve the use of toe clipping when performed only for the purposes of identification.

The Interagency Research Animal Committee considers toe clipping to be a painful procedure; however, it may be done only after IACUC approval with proper justification:

a) Toe clipping is done for unique and highly individualized animal identification that is permanent and unambiguous and is used for the long-term identification of potential breeding mice;

b) If the neonates need to be identified before day 7, toe clip is preferred;

c) If the same animal must be genotyped, then the animal may only undergo one procedure for both tissue harvest or genotype and permanent identification;

d) When no other individual identification method is feasible (such as ear notching, ear tags, tatooing, or subcutaneous implantation of a transponder identification chip);

e) Toe clipping for the sole purpose of obtaining DNA is not acceptable; and

f) The toe clip procedure should be detailed in the IACUC protocol.

Action

1. Toe clipping can be performed on pups up to 12 days of age; the preferred age is less than 8 days of age.  Toes may be removed without anesthesia on mice up to 8 days of age.  Toes must be removed with anesthesia on mice 8 days of age or animals whose eyes have opened and appropriate post-operative analgesics must be used.

2. The procedure should be performed with sharp, disinfected scissors.  Scissors must be disinfected between animals.  Research presonnel should confirm that there is no bleeding prior to returning the animal to its cage.

3. Toe clipping is limited to no more than one per foot.

4. Since the rodents hold food with their forepaws, the IACUC will only allow this procedure on hind paws, one toe per paw, and permits the removal of just the first bone.

5. The procedure is not recommended for rodents used in psychology research as it may affect the gait or grip strength of the animal.

References

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

2. IACUC Guidebook, 2nd Edition, NIH, Office of Laboratory Animal Welfare, 2002.

3. Good Laboratory Practice Regulations; Minor Amendment, Federal Register, VOl. 54, N. 75, 1989. (www.nal.usda.gov/awic/legislat/fdaclip.htm)

4. National Institutes of Health, ARAC, Guidelines for Toe Clipping of Rodents, Revised 5/12/04. (http://oacu.od.nih.gov/ARAC/).

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

Purpose

The Guide for the Care and Use of Laboratory Animals states that newly received animals should be given a period for physiological, psychological, and nutritional stabilization before their use. This allows animals to recover from shipping stress and permits them to adapt to their new surroundings. The length of time for stabilization will depend on the type and duration of animal transportation, the species involved, and the intended use of the animals. This policy will guide the amount of time an animal should be allowed to acclimate to the facility after their arrival from another institution.

Action

Non-Terminal Procedures

1. Rodent species should have a minimum acclimation period of 72 hours.

2. Non-rodent species should have a minimum acclimation period of 7 days.

3. Aquatic species should have a minimum acclimation period of 72 hours.

Terminal Procedures

1. For rodent species, an acclimation period of 48 hours is recommended, but is optional.

2. For non-rodent species, an acclimation period of 72 hours is recommended, but is optional.

3. For aquatic species, an acclimation period of 48 hours is recommended, but is opotional.

Terminal Training Procedures

No acclimation period is required for any species.

The PI should take into consideration the stress-induced variable in the research outcomes. For requirements for other unidentified  species, please contact the Center for Comparative Medicine Attending Veterinarian.

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

Purpose

In order to comply with government Regulations (USDA Animal Welfare Act and Animal Welfare Regulations), the UConn Health’s Institutional Animal Care and Use Committee (IACUC) has implemented a policy implementing a training form for individuals who are performing research, testing, or training on USDA-regulated species.

Action

1. All individuals who perform research, testing, or training on a USDA-regulated species are required to fill out, and maintain, an IACUC animal training records form.

2. This form is to be maintained by each individual using the regulated species.

3. Training forms must be attached to the protocol record if the protocol is an InfoEd Animal Use protocol submission.

Effective Dates: February 26, 2015 thru January 31, 2018

Purpose

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 (IACUC) has established a policy on requirements for reporting unusual or unexpected morbidity and/or mortality in a research project.

Action

1. If, during the course of a research project, any unusual animal deaths or significant health related problems occur, the Principal Investigator must report the incident within 72 hours from time of discovery to the IACUC (via the IACUC Coordinator) and the Director of the Center for Comparative Medicine (CCM). The Attending Veterinarian should also report any incidences of unusual animal illnesses and deaths to the IACUC via the IACUC Coordinator when they are discovered during review of animal morbidity and mortality reports.  This includes the discovery of any unanticipated pain, distress, or lethality of transgenic/gene-targeted animal lines.
2. It is at the discretion of the Attending Veterinarian what action, immediate or otherwise, is needed to prevent, reduce, or minimize any further animal suffering. This may include removal of the animal(s) from the study and/or euthanasia of the remaining animal(s).
3. There should be a combined effort of the IACUC, the Attending Veterinarian, and the Principal Investigator to arrive at a workable plan to prevent further morbidity and mortality for the duration of the research project.

Effective Dates:   December 1, 2022 through December 31, 2025

Purpose

In order to ensure appropriate use of Complete Freund’s Adjuvant (CFA), the UConn Health’s Institutional Animal Care and Use Committee (IACUC) has implemented a policy on the use of CFA.

Background

CFA is an oil-in-water emulsion containing mycobacterial cell wall components. It is used as an agent to potentiate antibody response to injected immunogens by eliciting a local inflammatory response. CFA should be used only when absolutely necessary (e.g., weak antigens, induction of auto-immune response) and may only be used for the initial immunization. Incomplete Freund’s Adjuvant (IFA) or other adjuvants that produce less intensive inflammatory responses (e.g., RIBI^® or TiterMax^®) should be used whenever possible. Improper use of Freund’s adjuvant can cause undesirable and painful side effects such as excessive inflammation and swelling, abscess formation, ulceration, and tissue necrosis.

Any proposed deviation from these guidelines must be fully explained and justified to the Animal Care Committee in the research proposal.

Occupational Health and Safety

Due to the fact that CFA contains killed mycobacteria, accidental inoculation into personnel handling this agent can result in subsequent positive tuberculin titers and/or local inflammatory lesions, which have poor response to antibiotic therapy. These lesions can be severe and result in granulomas, abscesses, and/or tissue necrosis. Extreme caution should be taken when using CFA.

Personnel Protective Equipment (PPE), especially gloves and eye protection, must be worn when handling CFA. Needles should not be recapped, Luer lock syringes should be used, and anesthesia or sedation of animals being injected is recommended to minimize the risk of accidental exposures to personnel.

Action

Procedure

CFA may be used only for the first (priming) immunization. IFA should be used for any subsequent, booster immunizations. Inoculum should be free of extraneous microbial or chemical contamination. Filtration of the antigen before mixing with adjuvant is recommended.

Sites of administration of the CFA should be chosen so as to avoid anatomic sites used for handling or restraint of the animals being inoculated. Intravenous administration of CFA is prohibited. Intramuscular, intradermal, and footpad administrations (rodents only, one footpad only) are permitted only with scientific justification and approval of the IACUC.

Injection sites should be clean and free of debris as contamination is likely to result in local infections. Clipping of hair and surgical preparation of the injection sites will reduce the potential for the development of infection and/or abscess formation. Sterile Luer lock syringes and needles should be utilized. Needles should be 25 to 31 gauge in size. The number of injection sites should be limited to the minimum necessary to accommodate the volume of material being administered. Injection sites should be sufficiently separated to prevent coalescing of any inflammatory responses from different injection sites.

Animals may receive booster immunizations of IFA or another adjuvant, as required, to develop the desired immune response as long as no adverse effects from prior immunizations are present. There should be a minimum interval of two weeks between initial and subsequent immunizations.

The following is a list of acceptable routes of administration and volumes of CFA for the indicated species:

Species	Subcutaneous	IM	SubQ via Footpad	ID	IP
Mouse	0.1 ml/site 4 sites maximum	NR (Not Recommended)	Requires Scientific Justification	0.05 ml/site 4 sites maximum	0.25 ml maximum
Rat	0.2 ml/site 4 sites maximum	NR	Requires Scientific Justification	0.05 ml/site 4 sites maximum	0.5 ml maximum
Rabbits	0.25 ml/site 4 sites maximum	0.25 ml/site 4 sites maximum Requires Scientific Justification	NR	0.05 ml/site 1.0 ml total	Not permitted

Animals must be closely monitored for complications following CFA administration. Daily observation of injection sites must be documented in written records for the initial 2 days post immunization and a minimum of once weekly thereafter. Injection sites should be closely monitored for the development of severe lesions. When the adjuvant has been administered intraperitoneally in rodents, the animals must be monitored closely for excessive abdominal distension.

References

1. ARAC Guidelines: Recommendations for Consideration in the Research Use of Inflammatory Agents. NIH Office of animal Care and Use. May, 1996.

2. Jackson, LR and Fox, JG. Institutional Policies and Guidelines on Adjuvants and Antibody Production. ILAR Journal, Vol. 37(3): 141-152, 1995.

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

Purpose

In order to comply with NIH-PHS government regulations which state that all vertebrate animal procedures described in a PHS grant must be approved by the institutional animal care and use committee, the University of Connecticut Health’s Institutional Animal Care and Use Committee (IACUC) has implemented a policy regarding new animal care and use protocol submissions.

Action

1. All animal care and use protocol submissions must be submitted electronically through the use of the InfoEd lab animal module.  The InfoEd electronic Research Administration (eRA) portal is web-based tool available at https://ovpr.uchc.edu/services/rics/animals/iacuc/protocol/.

2. Protocols must be submitted by deadlines which are on the IACUC website.  Only complete protocols will be reviewed by the IACUC; simply submitting a protocol by the deadline does not mean that the protocol is ready for committee review.  If your protocol is returned for any reason, you must resubmit the revised protocol by the deadline date to be considered at that month’s meeting.  Extensions of deadlines must be approved by the IACUC Chair.

3. Training on the use of InfoEd is highly recommended.  Information on how to receive training can be obtained from the Animal Care Committee office.

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