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The IACUC Unanticipated Adverse Event Form must be completed for reporting any adverse or unanticipated event affecting animals used in research, testing or teaching.

Principal Investigators should seek assistance from the Attending Veterinarian when adverse events or protocol deviations occur.  S/he can assist in assessing the situation, seeking resolutions, and helping with the report.  Consultation with the Attending Veterinarian MUST occur when pain or distress is beyond the level anticipated in the protocol description or when interventional control (such as administration of analgesics) is not possible.

Definition of an unanticipated or adverse event:  Any event not consistent with routine expected outcomes that results in unexpected animal welfare issues (death, disease, distress).

Examples of events that MUST be reported include, but are not limited to the following:

  • Animal death or illness from spontaneous disease not related to activities approved on a protocol.
  • Unexpected animal death or injuries related to approved animal activities (e.g., allergic reactions, broken limbs, complications during or recovering from surgery, sudden death). Unexpected death includes an increased number of deaths over what was stated in the approved protocol.
  • Death, disease or distress due to equipment failure or natural disaster.

The Adverse Event Form should be completed and submitted to the IACUC within 24 hours of observing the event.

Email a copy of the report to the IACUC Program Manager: Lauren Cantamessa,, (406) 994-6821.

Adverse events affecting USDA regulated species require a separate report for each affected animal.

Questions regarding the use of this form should be directed to the IACUC Chair, or the IACUC Program Manager.

Animal Adoption

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I. Purpose

The purpose of this policy is to allow research animals that are no longer assigned to protocols to be adopted by individuals as companion pets. The IACUC recognizes that adoption of research and teaching animals raises the potential for human health and safety concerns. While it is impossible to reduce health and safety risks to zero, animals selected for adoption must be healthy and present minimal risk of transmitting zoonotic infection (PI).

II. Background

The Animal Resource Center and Montana State University recognizes the invaluable contribution of research animals in the advancement of biomedical knowledge. The Institutional Animal Care and Use Committee (IACUC) endorses and promotes the responsible, humane, and appropriate use of research animals while complying with the concept of “refinement, reduction, and replacement” in our use of research animals. The IACUC also recognizes that sometimes animals finish a research or teaching project in good health but are not suitable or needed for any other projects at the institution. In such circumstances, the Animal Resource Center may consider finding adoptive homes for the animals. It is understood and widely accepted that adoption programs not only enhance the quality of life for healthy research animals that are no longer needed by the University, but can also decrease stress and raise morale for both the research and animal care teams. We believe that responsible and compassionate researchers utilize animals only when necessary; therefore it follows that we should do our best to ensure good lives for those animals whose sacrifice is not required by science.

III. Procedure

The following conditions must be met in order to place an animal for adoption:

1. Adoption privileges are granted only with the approval of the Attending Veterinarian.

2. An animal will be considered for adoption only if:

a. The purpose for which the animal was acquired no longer exists

b. The animal will not be needed in another IACUC approved study

c. The animal has been returned to the Animal Resource Center

d. An adopter has been identified to adopt the animal.

3. Before being released for adoption, animals must be examined by the Attending Veterinarian and found to be in good health and of suitable temperament appropriate for a pet. Cats and rabbits must be neutered and receive all age-appropriate vaccinations, as is accepted by standard veterinary practice. Cats may be declawed if requested by the adopting party with the understanding that the cat will remain an indoor pet.

4. The individual adopting an animal must sign a waiver which states the University is not liable for any injury or damage to persons or property by the adopted animal. The waiver further states that no warranties, guarantees, or promises of any kind have been made or can be made with regard to the adopted animal’s physical condition or temperament. The owner assumes all further responsibilities associated with responsible companion animal ownership.

5. An animal purchased from a Class A dealer (purpose-bred) may be eligible for adoption as long as the above conditions are met.

6. At the time of transfer of ownership, the adopter shall assume all financial responsibility for housing, care, and medication of that animal.

7. For each animal being adopted an “Animal Adoption Agreement” will be completed and signed by the attending veterinarian and the adopting party. This record should be maintained by the Animal Resource Center for a period of 3 years from the date of signature by the new owner.

8. Any deviations from the provisions of this policy must be approved by the Director of the Animal Resource Center and a simple majority of a quorum of the IACUC.  

IACUC Approval Date:  04/ 09/2014

Review Date: 04/ 09/2014

Issue Date:  04/ 09/2014

Animal Users and Principal Investigators Training Requirements

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I. Purpose

The purpose of this document is to describe training requirements for personnel working with animals and Principal Investigators (PI’s).

II. Scope

This document applies to all persons working with animals, or directly responsible for the conduct of animal research.

III. Process

A. Animal Users and PIs Prior to preforming any work on animals, the following training curricula must be completed.

1. Animal Care & Use Orientation

a. Occupational Health & Safety for Personnel Working with Animals

b. Biosafety for Personnel Working with Animals

c. Introduction to the IACUC

d. Animal Care at MSU

2. CITI species specific modules

3. Protocol specific procedure training and assessment

Note: Protocol specific procedure training and assessment must be documented and maintained by the PI.

B. Personnel working with animals housed in the ARC or personnel performing animal procedures in a laboratory.

1. Hands on species specific training as applicable

2. Procedure training as needed (e.g. injections, gavage, monitoring, etc.…).

C. Personnel Working with Nonhuman Primates must attend additional training covering:

1. Safety, risks and zoonoses associated working with NHPs

2. Herpes B virus and exposure kit procedures

 3. Tour of NHP facility, review of management and handling techniques and NHP behavior

D. Personnel Working with animals housed in JFLF

1. Facility orientation

2. Animal handling as applicable

E. PI Additional Requirements

1. Writing a Protocol- Core Elements

2. Protocol Review Procedures

3. PI responsibilities

4. PAM process overview

IACUC Approval Date: 04/18/2018

Review Date: 04/18/2018

Issue Date: 04/24/2018

Authority of the Attending Veterinarian

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I. Purpose

Montana State University (MSU) is committed to observing Federal policies and regulations and the Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC) International standards for the humane care and use of animals. This policy describes the authority of the attending veterinarian.

II. Scope

This policy covers all animals on MSU premises used for research, teaching, training, breeding, and related activities, hereinafter referred to collectively as “activities”, and is applicable to all persons responsible for conducting activities involving live vertebrate animals at or under the auspices of MSU.

III. Responsibility/Authority

The Institutional Animal Care and Use Committee (IACUC) is responsible for overseeing implementation of and ensuring compliance with this policy.

IV. Process

The Attending Veterinarian (AV) has been delegated authority to implement MSU’s veterinary care program, and to oversee the adequacy of all other aspects of animal care and use. Adequate veterinary medical care is essential to support a comprehensive animal care and use program that ensures the health and well-being of animals. The University has empowered the Institutional-appointed A (AV) with full authority to provide medical care or euthanasia, at their discretion, to animals on MSU premises. The duties of the AV may be shared or delegated to other veterinarians who directly communicate with the AV, but ensuring program compliance with federal regulations and institutional policy remains the responsibility of the AV.

The AV (or authorized designee) has unrestricted access to all areas where animals are used or housed, including any satellite facilities not directly managed by the Animal Research Center (ARC), and is authorized to immediately suspend research activities that are inconsistent with acceptable research conduct or for protocol deviations that have a negative impact on the animal. The AV will promptly forward incidents to the IACUC for further review.

The AV (or authorized designee) will make every reasonable attempt to consult the Principal Investigator (PI) prior to implementing veterinary care actions. However, in emergent situations, the AV is not required to obtain approval from the PI or other parties to provide appropriate care in an expedited fashion if such actions are deemed necessary by the AV to protect the interests of the animal or the integrity of the University’s animal care program. In such cases, the AV will notify the PI as soon as reasonably practicable.

If the responsible research staff member and the AV (or authorized designee) are not in agreement regarding the need for medical treatment or euthanasia, the AV (or authorized designee) has authority to act to protect the health and well-being of the animal and will make the final decision based on their medical judgment.

IACUC Approval Date: 10/17/2018
Review Date: 10/ 17/2018
Issue Date: 10/ 23/2018

Food and Water Restriction

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Background: Behavioral research often requires that an animal perform a task for which it receives food or fluid reward. This situation is not unlike conditions in the wild, in which animals must forage, travel distances, solve problems, or otherwise work to obtain food and water. Performing a task for rewards may also be behaviorally enriching for laboratory animals, especially nonhuman primates. However, a fundamental concern with studies that may involve food or water restriction is that animals are maintained in a healthy state and that they do not experience pain or distress. This document provides an overview of the issues and explains the IACUC’s policy regarding food or water restriction.

The IACUC referenced the following documents in setting this policy:

Guide for the Care and Use of Laboratory Animals (Eighth Edition). Washington DC: National Academy Press.

• NRC (2003). Guidelines for the Care and Use of Mammals in Neurosciences and Behavioral Research. Washington DC: National Academy Press.

The federal Animal Welfare Regulations (Code of Federal Regulations, Title 9).

• Toth, LA and Gardiner TW. Food and Water Restriction Protocols: Physiological and Behavioral Considerations. Contemporary Topics, 39(6). pg 9-17; 2000.

Definitions (Guide pg.30):

• Scheduled access: the animal consumes as much as desired at regularintervals.

• Restriction: the total volume of food or fluid consumed is strictly monitored and controlled.

• Fasting for surgical procedures: usually a period of less than 12 hours. This is not considered restriction and the following guidelines do not apply.


1. Individual animals may vary in their physiologic response to food or fluid restriction. Therefore it is imperative that animals be closely monitored on a daily basis to ensure that they are healthy, adapting normally and consume sufficient food and water to maintain their health status. Monitoring and if necessary, early intervention are the most important objectives of this policy.

2. It is the obligation of the investigator to demonstrate to the IACUC that food or water restriction is the only effective way to accomplish the scientific goals of the study. The objective when these studies are being planned and executed is to use the least restriction necessary to achieve scientific objective while maintaining animal well-being

3. It is the investigator’s ongoing obligation to continue to attempt methods of positive reinforcement that do not involve food or water restriction and to use restriction only when other methods fail.

4. The use of restriction to motivate behavior must be specifically discussed, adequately justified, and approved in each protocol in which it is used.

5. Criteria for monitoring animal health must be defined in the protocol (i.e. body weight, urine and fecal output, urine specific gravity, etc.). Criteria must be defined for temporary or permanent removal of an animal from the experimental protocol.

6. Investigators must maintain water consumption and food intake records in a form acceptable to the IACUC and readily available for inspection.

7. At the discretion of the IACUC, Investigators may be required to file updates or reports regarding the status of the animals on the restriction protocol.

8. At the discretion of the veterinarian, a particular animal’s food or water ration must be increased if:

a. The animal becomes ill or requires medical treatment.

b. The animal exhibits weight loss of greater than 10-15% of its initial body weight.

c. The animal shows evidence of clinical dehydration.

d. A young animal fails to show reasonable weight increase during a time when it should be growing.

e. The animal undergoes any procedure requiring anesthesia.

f. Under any other circumstance deemed necessary by the veterinarian.


Humane Experimental Endpoints

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The Guide for Care and Use of Laboratory Animals (8th Edition) defines the humane endpoint as the point at which pain or distress in an experimental animal is prevented, terminated, or relieved. The use of humane endpoints contributes to refinement by providing an alternative to experimental endpoints that result in unrelieved or severe animal pain and distress, including death.

The PI, who has precise knowledge of both the objectives of the study and the proposed model, is required to identify, explain, and include a study endpoint that is both humane and scientifically sound. The PI is also required to define the criteria for humane intervention for the study, listing how often the animals will be monitored to evaluate if they have reached the humane endpoint, and provide information on training of personnel performing the assessment.

When evaluating humane endpoints, the following specific sequela should be considered:

1. The procedure should not interfere with the ability of the animal to ambulate, eat, drink, urinate and defecate.

2. The procedure should not result in a net weight loss of more than 20% of the body weight.

3. The procedure should be ended if the investigator has conclusive evidence that untreatable organ failure has occurred, and the animal exhibits signs associated with the failure of the organ system.

4. Animals should be euthanatized if unrelated health conditions develop that make their experimental use of no value to the investigator.

5. Obvious signs of illness should serve as alternatives to death as an experimental endpoint.

The IACUC defines the moribund condition as a severely debilitated state that precedes imminent death. The use of death as an endpoint requires strong scientific justification in the Protocol and full committee review and approval by the IACUC.


IACUC Training & Continuing Education

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I. Purpose

The purpose of this document is to describe training requirements for IACUC members.

II. Scope

This document applies to all IACUC members.

III. Process

A. New Members

Upon appointment to the IACUC,   new members will be required to complete the following training.

1. CITI Training Essentials for IACUC Members

2. Protocol Review Process (provided by IACUC program manager)

3. Facility Inspection/Program Review (provided by IACUC program manager

4. Post Approval Monitoring (provided by IACUC program manager)

Note: IACUC members fulfill facility access requirements as applicable.

B. Continuing Education of IACUC Members

1. The IACUC Program Manager, IACUC Chair, or Attending Veterinarian are responsible for identifying and communicating educational opportunities available to the committee members such as webinars, workshops, and professional meetings sponsored by organizations (e.g.NIH/OLAW, PRIM&R, AALAS, SCAW, etc.).

2. In-house Continuing Education sessions will be offered periodically (typically quarterly). Continuing Education sessions may take place as part of a regularly scheduled IACUC meeting Examples of training topics include, but are not limited to:

a. Webinars: Mock protocol review and discussion.

b. Presentation and discussion of special topics, and/or emerging trends.

c. Presentation by IACUC members regarding sessions of interest attended at national or regional meetings.

d. Training sessions provided by outside consultants.

IACUC Approval Date:  04/18/2018

Review Date:  04/18/2018

Issue Date:  04/08/2018

Inter-Institutional Collaboration Requirements 

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Montana State University (MSU) is accountable for ensuring all work conducted meets applicable regulations. The intent of this policy is to provide guidance for review of projects involving collaborative work with live animals at outside organizations or in the field for the performance of research, testing, breeding or teaching.


This guideline applies to Principal Investigators, working with live vertebrate animals at an outside organization (Collaborating Organization) or in the field through an award, sub-award, contract or sub-contract from MSU.


Inter-Institutional Agreement (IAA): An agreement between MSU and the collaborating organization for the performance of animal research, testing, breeding or teaching.

Principal Investigator (PI): An MSU employee responsible for a proposal (i.e., “animal use protocol”) to conduct research and for the design and implementation of research involving animals.

Process Procedure:

1. PI is to provide the IACUC Program Manager with the following information:

a. Name of collaborating institution

b. Name of PI at collaborating institution, email and phone number

c. Research Project (grant/contract) title

d. Sponsor or funding agency if applicable

e. Sponsor’s award number if applicable


2. IACUC Program Manager responsibilities:

a. Initiation of the IAA through the PI at the collaborating institution. Assessment of the completed IAA for compliance and obtaining additional documentation as required.

1) Completion of an OLAW Interinstitutional Assurance (as applicable) if the collaborating institution does not have an OLAW approved Animal Welfare Assurance.

b. Distribution of the fully executed IAA and associated documentation as applicable to MSU PI and collaborating PI.

IACUC Approval Date: 03/27/2019

Review Date: 03/27/2019

Issue Date: 03/29/2019

Maximum Dosing Volumes

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  Adult Mice Rats(>250 grams)
Intramuscular (IM) 0.1 ml 0.3 ml
Subcutaneous (SC) 3.0 ml 10 ml
Intraperitoneal (IP) 2 ml  5 ml
Intravascular (IV) 0.25 ml 2 ml
Oral (PO) 0.5 ml 4 ml

Institutional Animal Care & Use Noncompliance Policy

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Montana State University’s (MSU) Institutional Animal Care and Use Committee (IACUC) ensures that animals used for research, teaching, training, or testing are treated humanely and that research is performed with the highest scientific and ethical standards. This policy establishes guidelines to address instances of regulatory or policy noncompliance by individuals engaged in the care or use of animals used for research, testing, training, or teaching. This policy is intended to address compliance issues that in the determination of the IACUC and in some instances, the Institutional Official (IO), go beyond a minor oversight.

Research and Activity Requiring IACUC Oversight

Federal laws and regulations require that all research, teaching, training, or testing involving live vertebrate animals have oversight by MSU’s IACUC. These animal related activities are subject to oversight by the United States Department of Agriculture (USDA) and the National Institutes of Health (NIH) Office of Laboratory Animal Welfare (OLAW) and must meet the standards set forth by the USDA Animal Welfare Act and Animal Welfare Regulations and the NIH Guide for the Care and Use of Laboratory Animals.

Reporting Suspected Noncompliance

MSU is committed to operating with integrity and in full compliance with all university policies and federal regulations. Suspected noncompliance violations may be reported by Principal Investigators (PI), laboratory staff, support staff, or the general public. MSU provides a number of avenues to individuals reporting a suspected noncompliance violation involving animal related activities. The phone numbers of contact persons including the IACUC Chair, Attending Veterinarian, and Director of the Office of Research Compliance (ORC) are posted in all animal facilities. Also, MSU has selected a private contractor, EthicsPoint, 855-753-0486, to provide an independent avenue for confidential reporting of a suspected noncompliance violation. All concerns will be treated as suspected noncompliance when initially reported, treated as confidential to protect all parties involved, and will be investigated promptly. MSU will not tolerate retaliation against individuals who report suspected noncompliance violations in good faith.

Examples of Noncompliance

Noncompliance with university policies or federal regulations can be classified as serious or moderate. Serious violations are the result of willful or malicious incidents relative to animal welfare, federalregulations, or violations that pose a real or potential threat to the health and welfare of animals. Moderate violations include instances where university policies are unclear, and which do not pose a threatto the health or welfare of animals.

Examples of violations include:

• Conduct animal-related activities without appropriate IACUC review and approval;

• Conditions that jeopardize the health or welfare of animals, including natural disasters, accidents, and mechanical failures, resulting in actual harm or death to animals;

• Failure to adhere to the standards set forth by the USDA Animal Welfare Act and Animal Welfare Regulation and the NIH Guide for the Care and Use of Laboratory Animals.

Investigation of Suspected Noncompliance

MSU will use the highest standards to investigate suspected noncompliance. Reported suspected noncompliance must be reported to the IACUC Chair and the ORC Director who will promptly initiate an investigation to gather facts to allow determination of the nature and extent of the concern,whether the issue presents a potential immediate animal health or welfare risk, and if the concern involves noncompliance with university policy or federal regulations. The involved individual(s) will be informed of the noncompliance investigation that is being conducted. If the IACUC Chair, in consultationwith the ORC Director, concludes that the noncompliance is serious or complex, a subcommittee may be appointed to conduct the investigation. The following considerations are evaluated during the investigation of the suspected noncompliance:

• Whether the reported actions resulted in jeopardizing the health or welfare of animals;

• Whether the animal-related activities were conducted without appropriate IACUC review and approval;

• Whether the reported violations constitute serious or continuing noncompliance with university policies or federal regulations.

When the investigation deems that noncompliance has occurred with university policies orfederal regulations, or that there is a past, present, or future threat to the health and well-being of animals, the noncompliance investigator(s) will providea report to the IACUC, ORC Director, and the IO. The reportshall include:

• A description of the noncompliance violation and whether the violation resulted in any adverse events.

• A summary of the records and evidence reviewed during the investigation.

• Identification of university policies or federal violations under which noncompliance occurred.

• Corrective actions that should be implemented to avoid noncompliance in the future and an appropriate date by which the corrective actions will be implemented.

Formal Determination of Noncompliance

When determination that a violation of university policy orfederal regulation has occurred, the IACUC Chair will notify the involved individual(s) in writing of thenoncompliance violation and corrective actions. In cases where the noncompliance is ongoing and presents risk to the health or well-being of the animal(s), the IACUC can suspend the research activity. If corrective actions are required, a timeline will be established in which the individual(s) must implement corrective actions. The individual(s) will have the opportunity to work with the IACUC, the AV, and the ORC Director to modify the corrective actionsif deemed appropriate by the IACUC. The Office of the Provost and the PI’s Department Head, College Dean, and the Office of Sponsored Programs may be notified of the noncompliance violation.

Examples of Corrective Actions After Determination of Noncompliance

Most moderate noncompliance violations that are not a result of willful or malicious intent and that do not pose a threat to animal health or welfare or violate federal regulations can be resolved administratively. For serious noncompliance violations, the IACUC maymandate remedial corrective actions. Such corrective actions may include, but are not limited to:

• Requiring specific training or retraining of the individuals involved in the proper care and use of animals;

• Additional monitoring by the IACUC, AV, or animal care staff of the animal-relatedactivities that pertain to the noncompliance violation;

• Requiring submission and approval of a IACUC protocol or a modification to an already approved IACUC protocol priorto continuation of the research for which noncompliance was reported;

• Restricting or limiting the scope of activities in which the individual(s) may engage;

• Suspending approval or terminating an approved IACUC protocol.

If any animal-related noncompliance is identified associated with an activity supported by the Public Health Service (PHS), the IACUC, through the IO, must promptly notify OLAW and the PHS funding agency, per PHS Policy, IV.F.3.

Reporting guidelines can be found at

Approved by IACUC 12-14-16 

Non-terminal Blood Withdrawal

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Species Maximum Blood Withdrawal
Mouse 0.1 ml per 10 grams body weight
Rat 1 ml per 100 grams body weight
Hamster/Gerbils 0.6 ml per 100 grams body weight
Rabbit 10 ml per kg
Nonhuman Primate 10 ml per kg


These stated volumes are the maximum blood to be withdrawn in a single bleed. At maximum, it can be repeated at two-week intervals. If more frequent bleeding is needed, the volume should be proportionally reduced. If more blood than this maximum volume is needed, the investigator must consult with the Veterinarian and receive IACUC approval.


Physical Restraint

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Physical restraint can cause both physical and psychological distress. According to the Guide for the Care and Use of Laboratory Animals (Eighth Edition), prolonged physical restraint should be avoided unless it is essential for achieving research objectives and is specifically approved by an Institutional Animal Care and Use Committee. If an animal will undergo prolonged restraint, training of the animal to the restraint device should be conducted to assure acclimation of the animal to the restraint device. Restraint devices are not considered normal methods of housing. As such, the MSU IACUC has adopted the following guidelines regarding the use of physical restraint in animals.

Definition: The use of manual or mechanical means to limit some or all of an animal’s normal movement for the purpose of experimental manipulation.


Restraint devices should be of suitable size, design, and operation to minimize discomfort or injury to an animal. The total time that an animal will have to be manually or mechanically restrained should be minimized. Methods for animal acclimation to the restraint device or activity must be defined. Animals that fail to adapt to the restraint device will be removed from the study. Animals must be observed at appropriate intervals, as approved by the IACUC. The veterinarian has the authority to temporarily or permanently remove any animal from the restraint procedures.

The following must be included in the animal care and use protocol: 

• Justification of the use of physical restraint in the context of the research objectives.

• The duration of restraint, which should be the minimum required to accomplish the research objectives.

• A description of the method and duration of time needed for training or acclimation to the equipment and personnel. Positive reinforcement should be used whenever possible.

• A description of monitoring methods.

• A description of endpoints for temporary or permanent removal from restraintmethods.


Post Approval Monitoring 

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I. Purpose

The purpose of this policy is describe methods for continuing IACUC oversight of animal activities as required by federal laws, regulations and policies. The Post Approval Monitoring program will enhance the IACUC’s ability to monitor approved procedural activities, provide information about programmatic risks and associated issues and convey practical corrective plans for the identified risks.

II. Scope

This policy applies to Principal Investigators (PI) and all individuals involved in research animal care and use at MSU.

III. Definitions

PAM: Post Approval Monitoring

IV. Process

PAM will be performed regularly to ensure that ongoing animal work conducted at MSU is conducted in accordance with IACUC approved protocols, institutional policies, and procedures. PAMs may be conducted for the following reasons;

A. Requested by the IACUC

B. Risk-based

C. Issue-based

D. Random selection

V. Procedures

PAM may be conducted utilizing a variety of methods depending upon the nature of the activities and goals of the monitoring. Typical methods for PAM may include but are not limited to all of the following activities:

A. Advance review of an approved protocol

B. Interviews with PIs, research, veterinary and husbandry staff

C. Comparison/verification of animal numbers

D. Reviews of training methods/records

E. Breeding colony management

F. Reviews of records for consistency with the protocol and institutional policies

G. Review of animal/use study areas 

H. Inquiries about unanticipated adverse events

I. Confirmation of appropriate medical surveillance participation

The findings of any type of PAM review will be shared with applicable individuals, the IACUC and regulatory oversight agencies as needed.


ILAR Journal, Volume 49, Issue 4, 1 January 2008, Pages 402–418

IACUC Approval Date:  05/16/2018

Review Date:  5/16/2018

Issue Date: 05/30/2018

Principal Investigator Criteria and Responsibilities

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 I. Purpose

The purpose of this guideline is to define the criteria for permitting individuals to fulfill the roles and responsibilities of a Principal Investigator (PI).

II. Scope

This guideline applies to PIs, and management responsible for identifying a new or replacement PI.

III. Definitions

Principal Investigator (PI): An MSU employee responsible for a proposal (i.e., “animal use protocol”) to conduct research and for the design and implementation of research involving animals.

IV. Qualifications/Eligibility

A. To be a PI, an individual must:

1. Be a faculty, adjunct faculty, or permanent staff of MSU who have earned a Ph.D. or other professional degree such as MD, DVM, DMD, DDS or similar.

2. The IACUC may utilize its professional judgement to approve an individual who does not meet the criteria above to serve as a PI.

3. Have an adequate understanding of the scientific rationale for the proposed work.

4. Have completed the appropriate animal facility access requirements.

5. Have completed required PI training.

V. Accountability/Responsibility

A. The PI is accountable for:

1. The development and obtaining IACUC approval of a protocol describing required experimental procedures and associated animal care and use, and ensuring active protocols are current through amendments and annual renewals.

2. Consultation with the Attending Veterinarian regarding husbandry, handling, medical treatment, immobilization, sedation, analgesia euthanasia, surgical planning, perioperative care and all procedures involving pain and/or distress for the development of protocols, .

3. Consultation with the Attending Veterinarian for amendments involving pain and/or distress. 

4. Ensuring the conduct of the research, testing and/or teaching involving the care and use of animals, following the methodologies described in the IACUCapproved protocol.

5. The care and use of animals in ways judged to be scientifically, technically and humanely appropriate (e.g. minimized pain and distress).

6. Reporting of events impacting or potentially impacting animal care, use and welfare.

7. Reporting deviations from approved protocols to the AV or IACUC.

8. The oversight of animal census associated with the approved IACUC protocol. This is accomplished in collaboration with the Animal Resources Center, the IACUC Administrator and/or any other appropriate Animal Resources personnel. Activities may include:

a. Transferring animals to another protocol (e.g., holding, training or active protocol)

b. Discontinuing, delaying or confirming standing orders for animals

c. Tracking animal numbers

d. Updating cage cards

e. Euthanizing animals

B. PIs must have the authority to provide the following assurances:

1. All activities proposed in the protocol:

a. Are in accord and consistent with the ILAR Guide for the Care and Use of Laboratory Animals and the Animal Welfare Act regulations (as amended), as applicable.

b. Comply with MSU policies, procedures and guidelines regarding the humane care and use of laboratory animals in research; unless appropriately justified and approved by the IACUC.

2. No activities involving live vertebrate animals will begin without an approved IACUC animal use protocol, or amendment as applicable.

3. All IACUC approved protocols will be kept accurate and up-to-date and will be submitted in a timely manner to ensure annual review and approval.

4. All experiments involving live animals will be performed under the PI’s direct or delegated supervision. The PI may delegate the hands-on animal work to qualified members of the research team. However, the PI remains ultimately accountable for the conduct of the research, testing and teaching procedures performed on animals as outlined in the IACUC-approved protocol.

5. All personnel conducting activities with the protocol will be trained in their respective responsibilities within the protocol.

6. All personnel conducting activities with the protocol must promptly report animals showing evidence of pain, distress or clinical health issues to a veterinarian.

NOTE: PIs must recognize that emergency veterinary care may need to be administered to animals showing evidence of pain or illness, or that euthanasia may be ordered by the AV for such animals.

7. A good faith effort has been made to search for alternatives to painful procedures in research animals, the literature search has been examined for potential alternatives and a written analysis of the utility of any cited relevant alternatives is provided in the IACUC protocol.

C. The PI is also responsible for:

1. Ensuring maintenance of complete and current animal study records in a secure accessible location. Records must be available at all times for review by the Attending Veterinarian, the IACUC, the USDA, FDA, and/or AAALAC.

2. Assuring all animals in the protocol are identified using appropriate cage cards.

3. Adhering to the IACUC Protocol Review Process.

4. Justifying the specific need for the use of animals (species and numbers) in the IACUC approved protocol.

5. Working with individual(s) during an internal audit of their protocol (e.g. IACUC Post Approval Monitoring). The PI is also responsible for completing any corrective action(s) assigned to him/her as a result of an audit within the given timeframe.

6. Providing information requested by the IACUC during the semi-annual Facility Inspection/Program Review. The PI is also responsible for completing any corrective action(s) assigned to him/her within the given timeframe.

7. Providing information requested by regulatory official(s) or AAALAC and answering questions if requested. 

IACUC Approval Date:  03/29/2018

Review Date:  03/29/2018

Issue Date:  05/08/2018

Prompt Reporting to OLAW under PHS Policy 

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The MSU IACUC will follow the guidance issued by OLAW, Office of Extramural Research on February 24, 2005. This guidance is intended to assist IACUCs and Institutional Officials in determining what, when, and how situations of noncompliance should be reported under IV.F.3 of the PHS Policy on Humane Care and Use of Laboratory Animals. PSH Policy, IV.F.3 requires that:

• “The IACUC, through the Institutional Official, shall promptly provide OLAW with a full explanation of the circumstances and actions taken with respect to:

o Any serious or continuing noncompliance with this Policy;

o Any serious deviation from the provisions of the Guide for the Care and Use of Laboratory Animals; or

o Any suspension of an activity by the IACUC.”

This guidance is available at: 

IACUC Approval Date: 09/30/2015

Review Date: 09/30/2015

Issue Date: 09/30/2015

Protocol Review

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I.      Purpose

To describe MSU’s IACUC procedures for reviewing new Animal Care and Use Protocols, changes to existing protocols (amendments), annual reviews (renewals); and to describe the process for submitting  a new Animal Care and Use Protocol and securing approval prior to expiration.

II.     Definitions

  • Protocol – A detailed plan or proposed scientific plan, treatment, or procedure to be conducted using animals.
  • Amendment – Any modification to an approved protocol that does not change the original scope of the protocol; classified as minor or significant.
  • Full Committee Review (FCR) – A quorum of the IACUC will review the protocol and vote to approve, return for modification to secure approval, or withhold approval.
  • Designated Member Review (DMR) – Following the opportunity to call for FCR, appointed IACUC member(s) reviews and determines the outcome for the assigned protocol/amendment.  Possible outcomes are return for modification, approval or request for FCR.
  • Expedited Review – Review of a protocol/amendment in a swift and thorough process due to critical time dependent events.
  • Administrative Approval – For minor amendments including, but not limited to, change to protocol personnel, and animal number increases up to 10%, the IACUC Program Manager and/or IACUC Chair determine the outcome.
  • Conflict of Interest (COI) – No IACUC member having association with the protocol under review or other COI may participate in the IACUC review or approval of a protocol/amendment, except to provide specific information requested by the IACUC.  A member who has a COI cannot contribute to the constitution of a quorum.

III.   Procedure

Protocol Review:

Principal Investigators (PIs) are required to consult with the Attending Veterinarian (AV) for all protocols, including protocols being submitted to replace a protocol that is expiring. After consultation with the AV, protocols can be submitted to the IACUC at any time using the IACUC Online Protocol system (IACUCOP).  Note: Every three years, an expiring protocol must be submitted as a new protocol. 

Depending on the nature of the protocol, some protocols can be reviewed soon after they are received by the IACUC office using the designated member review (DMR) process.  Any IACUC member may request FCR of a protocol at any time during the review process.

1.    Veterinary/Administrative Review

  1. Protocols can be processed through veterinary/administrative review prior to or concurrent with IACUC review. Allotted time: One calendar week
  2. Veterinary/administrative review of the protocol may be returned for Modification (RFM) to the PI by email notification prior to committee review. PI’s can revise and resubmit protocol.
  3. If the PI chooses not to address reviewer comments, a response to the RFM email including an explanation/rationale may be sent to the IACUC program manager.  The IACUC program manager will forward the email to the reviewer(s).

2.    Full Committee Review

Typically, the following protocols will undergo FCR:

  • Category E animals
  • Survival surgeries
  • Animal housing areas that are not part of the animal program overseen by the IACUC
  • Impact to personnel safety
  • Protocols involving USDA covered species
  1. Protocols scheduled for FCR are placed on the IACUC meeting agenda. Committee members receive electronic notification.
  2. All members have access to the protocol.  Comments/questions on the protocol may be submitted electronically.
  3. The protocol may be RFM to the PI via email prior to the meeting.
  4. The PI may be asked to present the protocol to the IACUC during a convened meeting (quorum required).  The presentation should include a brief overview of the study objective and address the recommended modifications/questions suggested by the committee. At this time, the IACUC can ask the PI any additional questions/ concerns, after which and the PI is excused from further meeting deliberation.
  5. The protocol will be discussed by the quorum of the IACUC.  After discussion the IACUC has the following voting options:
    • Approve
    • Return for Modification (RFM) and send to DMR (see process below) when resubmitted
    • Return for Modification and review during convened meeting
    • Withhold approval

3.    Designated Member Review

  1. The full committee is notified that a protocol will be assigned to DMR.  All IACUC members are given two business days to view/assess and comment on the protocol and the opportunity to request FCR. Allotted time: Two business days
  2. An IACUC member(s) will be assigned as the DMR(s) by the IACUC Program Manager following consultation with the Chair.  IACUC member(s) who are associated with the protocol or have a conflict of interest will not be assigned as the DMR(s). Allotted time: One calendar week
  3. The DMR(s) is required to review the protocol. If a protocol is assigned to more than one designated reviewer, the reviewers must be unanimous in any decision. They must all review identical versions of the protocol and, if modifications are requested by any one of the reviewers, the other reviewers must be aware of and agree to the modifications. Once the review of the protocol has been completed, the DMR(s) has the following voting options:
  • Approve
  • Return for Modification (to secure approval).  When a protocol is Returned for Modification, the following process occurs:
    1. PI is notified via e-mail of requested modification(s).
    2. PI revises and resubmits the protocol.
    3. Protocol is reassigned to the DMR(s) to verify the requested modification(s) have been addressed appropriately. The DMR(s) may RFM the protocol as many times as necessary to secure approval or may call for FCR (see IVA.2.b above). Allotted time: Two business days

B.    Amendment Review:

Amendments may be submitted at any time.  Amendments are reviewed in context to the original approved protocol which may necessitate review of the entire protocol but is not required to secure approval. Amendments may be reviewed by FCR (See III.A.2) DMR (See IV.A.2.b), Administrative Review or Expedited Review.

1.    Administrative Review

Minor amendments may be approved by the IACUC Program Manager for the following changes:

  • Addition or deletion of protocol associates
  • Up to 10% increase in non USDA covered species

Allotted time: Two business days

NOTE: This list is not inclusive.  IACUC leadership (Chair and Attending Veterinarian) may determine an amendment addressing other minor changes is acceptable for administrative approval.

2.    Expedited Review

Expedited review of an IACUC protocol amendment may be necessary, due to critical time-dependent events.

  1. The PI must obtain Department Head approval to request expedited review (email). To ensure that the expedited review process is executed in a swift and thorough fashion, the following items must be addressed prior to protocol amendment submission:
    1. If the proposed changes involve an increase in pain and distress, an appropriate literature search for alternatives to painful/distressful procedures must be conducted and included in the submission, and a consultation with the Attending Veterinarian or designee is required.
    2. When applicable, corresponding biohazard, particularly hazardous substances (PHSs) and radiation safety protocols must be in place.
  2. An email describing the rationale for expedited review; including the Department Head authorization memo must be submitted to the IACUC Chair.
  3. The IACUC Chair will notify the PI if the protocol amendment will undergo expedited review process as described below.
    1. Notification is sent to all committee members that the protocol will be assigned to DMR.  All IACUC members will be given the opportunity to view/assess and comment on the protocol, and the opportunity to request FCR. Allotted time: 24 hours
    2. Concurrently a DMR will be required to review the protocol. Allotted time: 24 hours

C.   Annual Review/Protocol Expiration:

Annual renewals  should be submitted at least 14 days prior to the renewal date to allow enough time for review and approval.

Protocols that are expiring must be submitted as an “original” protocol at least 30 days prior to protocol expiration to allow enough time for review and approval.

The IACUC provides reminder notifications of protocol expirations coming due at approximately 90, 60 and 30 days.  If a protocol has not been submitted 14 days prior to the protocol renewal date, the PI will receive a final reminder notification that if the protocol is not approved by the renewal date animal work described in the protocol cannot occur.

D.   Suspension

The IACUC may suspend an activity that it previously approved if it determines that the activity is not being conducted in accordance with the description of the activity by the principal investigator as approved by the committee.  The IACUC may suspend an activity only after review of the matter at a convened meeting of a quorum of the IACUC and with the suspension vote of a majority of the quorum present.

If the IACUC suspends an activity involving animals, the IO, in consultation with the IACUC, shall review the reasons for suspension, take appropriate corrective action, and report that action with a full explanation to APHIS and any Federal Agency funding that activity (if applicable).

Rodent Cage Density

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Guide for the Care and Use of Laboratory Animals, 8th edition TABLE 3.2 Recommended Minimum Space for Commonly Used Laboratory Rodents Housed in Groups*

Animals Weight (g) Floor area/Animala (sq. in) Heightb (in)
Mice in groupsc <10 6 5
  Up to 15 8 5
  Up to 25 12 5
  ≥25 ≥ 15 5
Female + litter   51 5

*The interpretation of this table should take into consideration the performance indices described in the text beginning on page 55. aSingly housed animals and small groups may require more than the applicable multiple of the indicated floor space per animal. bFrom cage floor to cage top. cConsideration should be given to the growth characteristics of the stock or strain as well as the sex of the animal. Weight gain may be sufficiently rapid that it may be preferable to provide greater space in anticipation of the animal’s future size. In addition, juvenile rodents are highly active and show increased play behavior. dOther considerations may include culling of litters or separation of litters from the breeding group, as well as other methods of more intensive management of available space to allow for the safety and well-being of the breeding group. Sufficient space should be allocated for mothers with litters to allow the pups to develop to weaning without detrimental effects for the mother or the litter.


Housing available at the MSU ARC for mice:

  Weight (g) Max # mice Tecniplast IVC (82 in2 ) Max # mice Alt Design IVC (75 in2 )
Mice in Groups ​<10  13 12
  Up to 15 10 9
  Up to 25 6 6
  >25 5 5
Female + Litter   1 1


 Breeding Schemes and Management:

Mice can be set up in trios, harems or pair breeding. Only 1 male is allowed per breeding cage. Multiple females may be bred to a single male; however, the total mice per cage may not exceed the cage capacity.

For trios and harem breeding the pregnant female should be separated and placed into her own cage (with appropriate nesting/enrichment materials) prior to giving birth. This is usually accomplished before E15 (15 days of gestation) when pregnancy is identified, or after mating when a plug is identified. If a female gives birth with in trio or harem cage, the male and remaining females should be removed to a separate cage leaving the female with her litter undisturbed. When pregnant females are separated, the male may remain with only one female or he can be moved to another breeding cage or housed separately based on the needs of the colony/protocol.

When breeding in pairs (one male to one female), the dam and sire may remain togethe r throughout gestation and lactation. Breeding pairs often breed during postpartum estrus immediately following parturition) so pairs with litters near weaning age must be monitored closely for the arrival of a new litter. Ideally, the current litter should be weaned just prior to the birth of the new litter; however if the new litter arrives early, the older litter must be weaned; even if it is not yet 21 days old.


Mice are usually weaned between 19-23 days of age with 21 days of age being the most common. They may be weaned as early as 17 days of age when made necessary by the death of the dam or the birth of a post‐ partum estrus litter. Litters may also be left with the dam for an extended time when underweight or small of stature, as long as the dam doesn’t give birth to another litter. Some transgenic, inbred, or specialty strains do not mature as quickly as normal wild type mice and require an extended nursing period. The weaning age is extended until they are mature enough to be weaned, and a notation is made on the cage label. When strains commonly require an older weaning age, this exception is noted in the animal use and care protocol. The weanling mice are separated by sex and housed in a density appropriate for the facility and caging. When genotyping, the IACUC policy Rodent Genotyping and Identification must be followed.


IACUC Approval Date:  03/29/2018

Review Date:  03/29/2018

Issue Date:  05/7/2018

Rodent Genotyping and Identification Methods

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Collecting Tissue for Genotyping and Identifying Purpose Bred Mice and Rats

The Institutional Animal Care and Use Committee (IACUC) must approve all methods for tissue collection prior to performing procedures on animals. Tail clipping and ear punching are acceptable methods of tissue collection for the purpose of genotyping mice and rats. The specific method(s) must be described and approved in the Animal Care and Use Protocol. Under all circumstances, aseptic method should be followed. Reference to this document is sufficient description, provided the guidelines are adhered to as described below.

Tail Clipping

This method involves amputating a very small segment of the distal tail. At 21 days of age or animals requiring a second tail sample must be appropriately anesthetized using ketamine/xylazine, ketamine/dexmedetomidine or isoflurane or local anesthetic. Animals >35 days of age that require tail clipping must be under general anesthesia using ketamine/xylazine, ketamine/dexmedetomidine or isoflurane during the procedures and administered a systemic analgesic (i.e., buprenorphine, meloxicam, carprofen) given at least once following the procedure. If multiple tail clippings are required a maximum of 1 cm total tail length can be amputated, with all tail clippings combined.

Ear Punch

This method involves punching a hole or making a notch in the ear pinna. Commercial ear punches are available and inexpensive. Ear notching using an ear punch is a permanent form of identification. Ear notch remnants can usually provide enough tissue for DNA sampling during the initial PCR screening. Ear punch samples collected on animals do not require the use of anesthesia or analgesics, however, for identification purposes the animal must be appropriately restrained to ensure proper technique. The ear punch device used must be disinfected between cages of animals. These devices can be autoclaved. 


Other Identification Methods

1. Micro Chipping: Injecting a small microchip transponder subcutaneously between the scapulae of the rodent. The microchip is detected by use of a reader.

2. Micro-tattooing: A permanent mark make using a needle and ink which is applied to the tail, toes, and foot pads.

3. Ear tagging: A metal tag with a unique identification number is attached to one ear of the rodent.

4. Non-toxic markers: Sharpies can be used to mark the tail or fur of rodents, however the mark must be reapplied every 24 hours to ensure the mark is still visible. Animal Marker is another product available which can be used on rodent’s fur. Animal Markers can last between 6-12 weeks.

IACUC Approval Date:  03/ 29/2018

Review Date:  03/29/2018

Issue Date:  05/07/2018


1. Guide for the Care and Use of Laboratory Animals, 8th Edition pg. 75

2. Hankenson FC Laire, Garzel LM, Fischer DD, Nolan B, and Hankenson KD. Evaluation of tail biopsy collection in laboratory mice (Mus musculus): Vertebral ossification, DNA quantity, and acute behavioral responses. J Am Assoc Lab Anim Sci 47:10-18, 2008

Reporting Animal Welfare Concerns (Whistle-Blower Policy)

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I. Purpose 

The purpose of this guideline is to describe the methods for reporting animal welfare concerns. The use of animals in biomedical research is necessary and appropriate. Montana State University is committed to the humane treatment of animals and strives to ensure that institutional facilities and procedures adhere in all respects to NIH guidelines for care and use of laboratory animals.

II. Scope

The IACUC wants to be informed of any concerns that individuals of the campus community may have with respect to the care and use of laboratory animals and feels it is important that these concerns be addressed on an individual basis.

III. Process for Reporting Animal Welfare Concerns

Principal Investigator (PI): An MSU employee responsible for a proposal (i.e., “animal use protocol”) to conduct research and for the design and implementation of research involving animals.

Any individual who witnesses what he/she perceives to be inappropriate treatment of research animals has several options to report the inappropriate treatment.

a. Contact any member of the IACUC and report the concern.

b. File a concern by filling out an “Animal Concern Form” (posted at all locations where animals are housed), which will be submitted to the IACUC.

c. MSU has instituted an anonymous Compliance Hotline via a third party company, Ethics Point that allows the reporter to remain anonymous. A report can be submitted electronically or by phone 855-753-0486.

Every effort will be made to keep the identity of the person raising the concern confidential. Retaliation against a person who raises a good-faith concern will not be tolerated. The IACUC will follow-up on all complaints and gather additional information as necessary and appropriate. If indicated, corrective action will be initiated and the results of the investigation will be communicated to the individual who raised the complaint.

IACUC Approval Date:  09/30/2015

Review Date:  09/30/2015

Issue Date:  09/30/2015

Use of Non Pharmaceutical Grade Compounds 

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I. Purpose

The purpose of this IACUC policy is: To communicate the IACUC’s expectations regarding the use of materials in or on animals. To provide guidance to endusers to enable them to be compliant with these expectations. To prevent or minimize potential toxic or unwanted side effects associated with the administration of non-pharmaceutical grade materials to laboratory animals.

II. Scope

Investigators are expected to use pharmaceutical-grade compounds whenever they are available4, 5,6,7,8, to avoid toxicity or side effects that may threaten the health and welfare of the animals and/or interfere with the interpretation of research results. Non-pharmaceutical grade compounds should only be used after specific review and approval by the IACUC of the written justification for use in an animal protocol. 

III. Definitions

A. Pharmaceutical grade compound - an active or inactive drug, biologic or reagent for which a chemical purity standard has been established by any recognized pharmacopeia, such as the: US Pharmacopeia (USP)/National Formulary (NF), British Pharmacopeia (BP), or Pharmacopoeia of the Council of Europe (EP). These include, but are not limited to, pharmaceutical compounds approved for human or veterinary use approved by the U.S. Food and Drug Administration (FDA).

B. Chemical Purity Grades – the majority of chemicals are manufactured to comply with the International Organization for Standardization (ISO) regulation ISO 9001:2008, and laboratory chemicals manufactured to standards set by the American Chemical Society (ACS).

C. Biologics – biological molecules, obtained either by collection or extraction and purification from living systems, or by production in recombinant expression systems, or by de novo chemical synthesis. In terms of FDA licensed products, examples include antitoxins, antivenins, blood, blood derivatives, immune serums, immunologic diagnostic aids, toxoids, vaccines, and related articles.

D. New Investigational Compound - supplied by its manufacturer for testing in an experimental setting only and for this reason would not have chemical purity standards established; by default is considered a nonpharmaceutical grade compound.

IV. Process

The appropriate chemical properties of a non- pharmaceutical grade compound should be considered by the Investigator for the proposed study and route of administration. The grade/purity, potency, concentration, pH, osmolality, stability, formulation (buffer or solvent), and potential contaminants (e.g. chemical, biological, and microbial, including pyrogenic substances), as well as handling and storage procedures, are among the properties and practices that impact quality of the compound for achieving the scientific aims of the study.

Scientific justification for the use of non-pharmaceutical compounds may include the following reasons9:

A. Non-availability of an equivalent veterinary or human drug

B. Specific exception to an available veterinary or human drug, for example:

1. Insufficient strength of the active compound in an available formulation

2. Presence of an excipient or preservative is unacceptable for proposed studies (e.g. toxic via planned administration route, nature of excipient would affect experimental model or compromise data collection)

3. Use of an available formulation requires further change (e.g. dilution or other addition) and offers no advantage over formulation from a high-quality reagent

C. Scientific necessity for comparability to previous research or to replicate specific experimental model

4 9 CFR Chapter1, Subpart A, sections 1, 2, and 3 5 US Department of Agriculture /Animal and Plant Health Inspection Service - Animal Care Policy #3 Veterinary Care, March 25, 2011. 6 PHS Policy on Humane Care and Use of Laboratory Animals, Guide for the Care and Use of Laboratory Animals, 8th edit. 2011. 7 Office of Laboratory Animal Welfare: LAW Position Statement: Non-Pharmaceutical Grade Substances: 8 Association for Assessment and Accreditation of Laboratory Animal Care International, FAQ 9 Cost savings is not included as an appropriate justification for use

IACUC Approval Date:  04/18/2018

Review Date:  04/18/2018

Issue Date: 04/25/2018

Xenopus and Zebrafish Larvae Policy

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Does the PHS Policy apply to larval forms of amphibians and fish?

Yes, larval forms of fish and amphibians have vertebrae and are covered by the PHS Policy. As noted in FAQ A4, the PHS Policy applies to the offspring of egg-laying vertebrates only after hatching. Zebrafish larvae, for example, typically hatch 3 days post-fertilization.


Zebrafish (Danio rerio): General Guidelines (Adapted from NIH Guidelines):

Current OLAW interpretation of PHS policy considers aquatic species as "live, vertebrate animals" at hatching. Although this is an imprecise stage for zebrafish it can be approximated at 72 hours post fertilization. For purposes of accountability all stages of development greater than three days of age should be described in an approved MSU IACUC Animal Use Protocol. An estimate of the number of larval zebrafish from day 4-7 days post fertilization (dpf) should be included.

Since these early stages (4-7 dpf) do not to feel pain or distress, it is preferable that their numbers be separated from zebrafish ≥8 dpf. This number can be listed as Column C in the Pain and Distress Category as a separate number from zebrafish ≥8 dpf.

The pain and distress categorization of the ≥8 dpf fish should be determined by the investigator based on the specific procedures described in the protocol. The number of animals used may need to be provided as an estimate, particularly with these young larvae, considering their size and normal housing conditions.

African Clawed Frogs (Xenopus spp.)

For xenopus, the beginning of the hatching process for most begins at about 2 dpf (50 hours post fertilization). The hatching process is completed over the next 48 hours and they begin to feed (4 dpf). This corresponds approximately with Nieuwkoop and Faber stage 45. For purposes of accountability all stages of development starting at 4 days of age should be described in an approved MSU IACUC Animal Use Protocol.

Since the brain is not fully developed until stage 53 (approximately day 24) it is likely that they cannot perceive pain. Larvae prior to this stage can be listed as Column C in the Pain and Distress Category.

The pain and distress categorization of the ≥stage 53 xenopus should be determined by the investigator based on the specific procedures described in the protocol. The number of animals used may need to be provided as an estimate, particularly with these young larvae, considering their size and normal housing conditions.


IACUC Approval Date:  09/19/2018

Review Date:  09/19/2018

Issue Date:  09/21/2018



  1. University of Oregon (2008) Final Report to OLAW on Euthanasia of

  2. National Institutes of Health (2009) Final Report to OLAW on Euthanasia of


    a. Guidelines for Use of Zebrafish in the NIH Intramural Research Program

  4. Babošová, P. Vašeková, J. I. Porhajašová & J. Noskovič (2018) Influence of temperature on reproduction and length of metamorphosis in Xenopus laevis (Amphibia: Anura), The European Zoological Journal, 85:1, 151-158,

  5. Xenbase

  6. Carlana Ramlochansingh, Francisco Branoner, Boris P. Chagnaud, Hans Straka

    a. Efficacy of Tricaine Methanesulfonate (MS-222) as an Anesthetic Agent for Blocking Sensory-Motor Responses in Xenopus laevis Tadpoles; Published: July 1, 2014