Instructions for Completing the MSU-IACUC Protocol Review Form

Forms, instructions, policies, and technical information regarding approval to conduct projects using vertebrate animals are available through the MSU Regulatory Compliance web site http://www.msstate.edu/dept/compliance.

The MSU Institutional Animal Care and Use Committee (IACUC) uses information you enter onto this form to evaluate projects using vertebrate animals in teaching, testing, and research. Careful review and completion of this form is essential if IACUC members are to understand accurately the many aspects of your study. The IACUC, in most reviews, does not use original proposals, standard operating procedures, or other sources to augment its review. Therefore, pre-submission review and a possible personal appearance at the IACUC meeting are the usual methods used to augment or clarify contents of the Protocol Review Form.

Name of Principal Investigator(s): At least one PI must be a faculty member at Mississippi State University and must have a significant role in the project.

Title of Project: The title should reflect the theme or content of the work intended. Short titles are preferred.

The Checklist: Read this list of items carefully. Failure to adhere to all rules or steps listed may prevent a review. All questions must be checked or answered.

Note: Submission deadlines and meeting schedules are available on the MSU Regulatory Compliance web site.

Note that (1) The Protocol Review Form must be reviewed by an IACUC member-representative prior to submission to the IACUC. (2) The questions require a literate answer. (3) An Occupational Health and Safety document must be submitted with the protocol (Risk Inventory Form) and to the Student Health Center (Health Questionnaire).

The paragraph at the bottom of the page invites you to discuss your protocol before the assembled committee. The usual format is for you to review briefly the project components and for IACUC members to ask questions to clarify points. You will not be present for the IACUC's discussion and vote.

PART I - PROJECT DESCRIPTION

1. Introductory Information: Provide this information as requested. Be certain that all participants having contact with living or dead animals or animal parts or data review and analysis are listed and their participation and training status indicated.

2. Project Information: Check if this is an initial (new, first time) submissions of this project to the IACUC, or if it a revision or rewriting of a previous protocol (give approval number), or if it is simply a renewal of an essentially unchanged but previously approved project (give approval number of earlier protocol).

Project Title: Repeat the title given on Page 1.

Indicate the primary category of protocol being submitted: research, teaching, or other [testing, generic (for techniques used often), or "umbrella" (including many specific protocols to be submitted)].

Note that research protocols must be rewritten on a current form every 3 years, unless significant revisions are needed or numerous amendments will not allow thorough presentation and understanding of the work intended.

Provide the project period (entire time period for the project's in vivo, in vitro, and the preparation and review periods) and then the actual time living animals will be used in this project.

Provide also the source of the funds and budgets to be billed.

3. Project Summary: This "lay" summary must describe, in terms understandable to a nonscientist (8th grade level), how you intend to use the animals in the project. Describe especially any procedures likely to cause pain and/or distress.

PART II - ANIMAL PROTOCOLS

1. Hazards: The IACUC cannot review a study using vertebrate animals unless the committee members are confident that both humans and other animals are protected from possible harm. Evidence that proper precautions will be in place requires definitive approval by one or more of the review committees listed.

Approval of the project by a hazard oversight committee will then allow IACUC review, but the IACUC may impose additional restrictions on hazard containment in accordance with regulatory and accreditation mandates.

2. Proposed Animals:

A. Species: Give the species or common name of the animal or animals involved in the study.

B. Source: Complete for each species. If more than one species, write the name of that species adjacent to the appropriate line.

A "dealer raised" dog or cat dealer breeds and raises the animals at his or her own facility. The animals' complete history is known. A "random source" dog or cat dealer obtains animals from many sources and has only to hold the animals for 5 business days.

Purchasing or otherwise obtaining animals from non (USDA) licensed sources poses certain concerns, so these intended sources must be reported to the IACUC.

Animals may be collected from natural habitats, but appropriate permits must be obtained before collection. Animals may be obtained also from MSU breeding colonies (e.g., farm animals, rodents, and fish).

Animals may be transferred among projects (recycled), but the IACUC is sensitive to the total history of an animal's use and will determine when multiple use impairs an animal's well being. At that point the animal will be removed from the research-teaching pool.

C.- L.Complete as requested. If the dimensions of the animal's primary enclosure (cage, stall, and/or pen) are not in accordance with the size, weight, and/or number of animals housed in that enclosure. See the Guide for the Care and Use of Laboratory Animals (1996) or the Guide for the Care and Use of Agricultural Animals in Agricultural Teaching and Research (1999) for allowed densities/cage sizes. Housing animals in "undersized" cages requires a request for an IACUC exemption. The use of wire bottom cages for housing rodents requires special IACUC permission/exemption.

3. Number of Animals: Place the optional number of live animals to be used in Category A if the animals are being held for anticipated future list. If more than one species is to be used, list all species and number of each to be used. You may indicate the number as total for the entire project duration or as number to be used per unit time, but in no case except by protocol amendment can the number(s) of animals be increased without prior IACUC approval.

Place the number of animals in Category B1 if animals are to be used in teaching, testing, or research and are to experience 1-3 instances of transient pain and/or distress within a period of hours to a few days only. Examples of Category B. procedures are:

a. Simple blood sampling.

b. Rectal examination.

c. Vaginal smear sampling.

d. Force feeding of innocuous substances.

e. Radiographing unanesthetized animals.

f. Terminal experiments under anesthesia.

g. Restraint in pens (but animals can stand or lie down).

h.Immunization using (non-Freund's) adjuvant.

i. Administration of drug.

j. Short term mechanical or manual restraint.

k.EEG, EKG, EMG administration.

l. Short term aversive stimulus application.

m. Early tumor growth.

n. Pithing frogs and fish.

o. Minor surgical procedures using local anesthesia and analgesia.

p. Short term food or water deprivation.

Place the number of animals in Category B2 if any procedures listed above are repeated more than 3 times.

Place the number of animals in Category C1 if the procedures intended may cause pain and/or distress and for which anesthetics (general or local), analgesics, or sedatives-tranquilizers are used to relieve that pain and/or distress or to facilitate restraint even though the procedure itself intended is not painful. Category C1 is for animals that experience single or widely spaced instances of pain and/or distress for which drugs are used. Examples of Category C procedures include:

a. Frequent blood sampling.

b. Pyrogenicity testing.

c. Insertion of indwelling cannulae/catheters.

d. Use of casts (plaster of Paris).

e. Immobilization/restraint (e.g., primate chairs, inhalation chamber).

f. Skin transplantation.

g. Cesarean section.

h. Recovery from anesthesia.

i. Immunization using complete Freund's adjuvant.

j. Surgical procedures with pain or recovery.

k. Moderate food or water deprivation.

Place the number of animals in Category C2 if the potentially painful procedures listed above are followed by a lasting (beyond one week) pain and/or distress not routinely alleviated with drugs. Examples of painful procedures that may be placed into Category C2 (or D) include:

a. Collection of ascetic fluid.

b. Total bleeding without anesthesia (excluding decapitation).

c. Production of genetic defects (e.g., muscular dystrophy, hemophilia)

d. Prolonged deprivation of food, water, or sleep.

e. Immobilization with muscle relaxants.

f. Some experimental infections.

g. Carcinogenicity research with tumor induction.

h. Application of painful stimuli, induction of convulsions.

i. LD50 and LC50 testing.

j. Immunizations of footpad or with adjuvant.

k. Shock or burn, organ transplant rejection studies.

l. Severe trauma.

Place the number of animals in Category D when the animals will experience unrelieved pain and/or distress for more than a transient period. Procedures inflicting continuing, unrelieved distress on vertebrate animals must be justified thoroughly in writing.

4. Experimental Design:

A. Explain clearly and concisely why living animals must be used in the study and why the species selected is/are the most appropriate for the study intended.

Where controls are appropriate, they must be described. Only in exceptional circumstances will the IACUC comment on experimental design, unless as the design indicates error or misjudgment.

B. The IACUC members must understand the basic design of your study and how experimental and control groups are constituted.

C. An appropriate number of animals is essential. Using too few or too many is wasteful. It is advisable to include extra animals (e.g., 5-10%) in case of death or experimental error; however, be prudent with numbers added. Any modification to the number of animals approved in the original protocol must be submitted as an amendment to the IACUC and approved prior to implementation. Some further concerns are:

1) If you are not competent with statistical analysis, a statistician must be consulted regarding required sample sizes. The statistician, if consulted, must sign and date the form. As an alternative, numbers may be based on published work that resembles closely the work you propose to do.

2) Be certain that the numbers of animals proposed are consistent throughout the protocol. If you discuss different treatments, be certain that the sample sizes in each treatment add to the total number of animals indicated elsewhere.

3) If you do not know the exact number of animals involved (such as in a field-oriented research project), then estimate the maximum number of animals you intend to use, test, or observe. Excessively high or low numbers of animals used should have a sound justification.

5. Care and Use of the Animals:

A. Technical Procedures

Describe concisely what experimental procedures (e.g., injections, surgery, sample collection, telemetry attachment, behavioral modification, or disease induction) will be done with the animals.

B. Assessing pain, stress, or distress

1) Restraint

a. Describe any restriction of an animal's movement (other than that imposed by its home cage, stall, or pen): manual, continuing, or chemical that is part of the study.

b. Some confinement-type restraint devices (e.g., metabolism cages, towel wraps, rabbit boxes, cat bags) may on initial use cause such stress in experimental animals that careful desensitization to the device is essential before the actual study procedures begin. This section asks for details of such procedures.

c. Monitoring animals regularly in longer-term (over 2 hours) confinement is essential to their well being.

2) Distress without pain

Pain is not the only criterion in humane considerations. Many distressful situations may not involve pain. Consider here distressing situations, if any, present in your study.

3) Minor Invasive Manipulations

a. Minor procedures were listed previously. This section asks you to describe if any means will be used to alleviate that minor pain and/or distress. It is not mandatory that substances be used in this case, but they should be considered because some minor procedures may be very painful.

b. Adjuvant Use: If you are going to use adjuvants to enhance an immune response, describe the products and procedure.

4) Substance Administration

a. IACUC members must be informed of the effects on an animal's well being of substances administered. The information you provide in this section supplies that information. The IACUC will not necessarily deny use of harmful substances, but the members must know the nature and probability of the harm.

b. If the substances' components for legal reasons cannot be provided to the IACUC, then this assurance of providing the confidential information to an appropriate member of the committee may satisfy the entire committee's need to know.

5) Nutrient Restriction

a. Describe if food or water will be restricted and why. Animals must not become excessively thin or obese. Food deprivation to increase motivation for food awards is allowed without special IACUC permission but only down to 80% of average maintenance intake, but such restriction still must be described.

b. Animal well being depends in part on proper nutrition. Indicate either if the diet will be nutritionally adequate or not, and, if not, explain the reasons for using the diet and possible adverse consequences.

6) Surgery: Anyone intending to complete this section must review the IACUC document "Surgery Guidelines for MSU Owned Animals" available on the Regulatory Compliance web site.

a. A "major" procedure involves entering a body cavity with any incision larger than that caused by standard sizes of hypodermic needles; conducting surgical procedures about the head, neck, perineum, or joints; or causing through surgery a residual disability.

b. The IACUC must determine that participating personnel (if observers of students) are qualified to conduct humanely the specific procedures involved.

c. Preoperative food and water restrictions must be noted here.

d. Multiple, major, survival surgical procedures are in most cases not allowed without good scientific reason. Economic considerations, such as "using fewer animals," is not an acceptable reason for multiple survival procedures on single animals. The sequential procedures must be related one to the other to fulfill the goals of the study.

e. The alleviation of pain and/or distress in animals undergoing major surgery (terminal or survival) is essential. Provide details regarding drugs used to alleviate pain and/or distress.

f. Non-drug methods (e.g., companionship, movement restriction, and heat) may be used also to make an animal less anxious and painful.

g. Describe presurgical considerations that would ensure a successful outcome for the surgery.

h. Describe in some detail the surgical procedure to be done. This may duplicate or expand on information given under Technical Procedures above.

i. State where the surgery will be done and where the (detailed) anesthesia and surgery records will be maintained (for ready access).

j. Neuromuscular blocking agents must be used carefully before and during surgical procedures.

1. Provide justification for the use of a neuromuscular blocking agent.

2. Describe how an animal that receives a neuromuscular blocking agent will be monitored for evidence of pain and/or distress.

k. Documentation of postoperative recovery is reviewed carefully by regulatory agencies, and provision of this information is essential.

7) Prior Use of Animals: The IACUC must ensure the well being of animals throughout their use by the university. The committee will determine from information provided in this question whether or not an animal may continue to be used as a research or teaching subject and in which ways.

8) Unintended Consequences: Unless the procedures proposed are unique and not related to other procedures, you should have some idea of what might "go wrong" with one or more of the techniques intended that would cause unintentional or undesired consequences, usually involving pain and/or distress. The IACUC should be informed of these possibilities so this and further studies of its type can be evaluated more accurately and the pain and/or distress category adjusted.

9) Euthanasia and Disposal:

a. Consult the 2000 AVMA document before describing how study animals will be killed. Conditionally acceptable methods must be justified in writing and an IACUC exemption requested. Describe drugs used and how they are administered. Describe how persons conducting euthanasia will be trained (and monitored, if need be).

b. If the animals in the project are likely to die or endure prolonged pain and/or distress, describe here the characteristics (endpoint) of an animal that will be killed before death or a state of extreme pain and/or distress is reached.

c. Describe what will become of the live animals after you are done with them.

10) Medical Care of Animals: Several veterinarians provide veterinary care for their research animals, whereas some researchers utilize others' expertise. Name the veterinarian(s) who will provide medical care for the animals on this project. Make certain that veterinarian is competent in the procedures to be done.

11) Reduction, Refinement, Replacement, and Literature Searches:

a. Database Search: This section must be completed with careful attention to questions asked and information required. Note that a detailed answer is required only for use of certain warm-blooded mammals (dogs, cats, rabbits, guinea pig, wild mammals, farm animals in biomedical research, hamsters, and some others) or for vertebrates on studies involving U. S. government funds when those animals are used in studies involving more than momentary pain and/or distress (even when anesthetized). You must, to comply with Federal law, describe in detail the literature consulted, the results or conclusions obtained from the search that verify that non-animal or less painful alternatives are not suitable or available, and that this work was not done previously.

b. Details of literature search: You must answer the four questions given under this section to assure IACUC members that you have considered alternatives to pain and animals and that you are not duplicating "excessively" other work.

c. Attending Veterinarian: The "attending veterinarian" is a legal entity which is Dr. Lucy Senter. This person may be contacted by email (lsenter@research.msstate.edu) as an alternative to obtaining an actual signature, but she must review protocols in the categories named in Item a. just above.

PART III - PERSONNEL TRAINING

1. Personal Qualifications: This section also requires special attention. Persons participating in the protocol's procedures must be competent in their tasks/techniques or supervised directly by someone who is competent in the tasks/techniques. Answer this set of questions separately for each person on the project.

Training has several components, including attending the basic survey class, reviewing educational materials available from the Office of Regulatory Compliance, and "hands-on" and "on-site" training by individuals trained previously. Statements that an individual holds a certain academic degree or diplomate status is not considered adequate evidence of qualification to do the specific tasks in this study.

2. Occupational Health and Safety: Follow the steps required, which include annual submission directly to the Student Health Center of a Health Evaluation Form for each participant and with each protocol a Risk Inventory Form for each person involved.

3. Certification: The Principal Investigator(s) must read the conditions and certify adherence to those conditions with a signature.