Office of the Vice-President, Research and Innovation Animal Use Protocol Form (AUPF) Instructions Under the Ontario Animals for Research Act, (Revised Statues of Ontario, 1990, Chapter 22 as amended by 1994, c27, s9; 1997, c41, s115; 1999, c12, Sch. A, s3; 2005, c2, s2 and Regulations 16, 17, 18, 19. Revised Regulations of Ontario, 1980, March 1990), and the University of Toronto Animal Care Policies and Guidelines, it is a requirement that all researchers who will be conducting research, testing or teaching projects at the University of Toronto involving the use of animals obtain research ethics approval before commencing the project. The approval process starts with the submission of an Animal Use Protocol Form (see www.research.utoronto.ca/forms/aupf/ ) to the appropriate Local Animal Care Committee (LACC) (see www.research.utoronto.ca/about/boards-and-committees/local-animal-care/ ). Please note that the AUPF was updated in January, 2012. Older versions of the protocol form will not be accepted after May 1, 2012. The version date is indicated on the bottom right corner of the AUPF. General Instructions 1. The protocol must be typed. Handwritten protocols will be returned. 2. Please complete all sections of the protocol. Insert N/A where the requested information does not apply. Incomplete protocols will be returned. 3. Submit one signed original and one PDF copy for review to your LACC. Review Local Animal Care Committees (LACC) for meeting dates and submission deadlines (see www.research.utoronto.ca/about/boards-and-committees/local-animal-care/ ). 4. If a proposed project involves more than one (1) animal species, a separate protocol for each species must be completed (the only exception may be for the use of invertebrate species contact the Chairperson of your LACC or the University Veterinarian prior to the preparation and submission of a protocol involving invertebrates). 5. There must be an approved protocol for all procedures involving the use of vertebrate and higher form invertebrate animals (e.g. cephalopods, etc.) being used by faculty members, graduate and undergraduate students, research associates, and all other personnel regardless of source of funding. Protocols for non-faculty members must be submitted under the name of the senior faculty member supervising the project. 6. Any animal research or teaching project administered by the University which is to be conducted in the field or in a facility not owned by the University of Toronto must be covered by an approved protocol. 7. The form must be submitted under the name of a faculty member. Animal Use Protocol Form (AUPF) Instructions Page 1 of 8
Modifications a) Any MAJOR modification to an approved project in progress (e.g. the inclusion of new procedures involving potential pain, surgery or anaesthetization, or a change in Principal Investigator or course director) must be described in a University of Toronto Animal Use Protocol (which can be completed but not saved on your computer). This protocol must be signed and approved in the manner described. b) Any MINOR modification to an approved project in progress (e.g. changes in number of animals to be used, location of experiment or changes in personnel) can be requested by completing the University of Toronto AUPF Amendment Form (available from www.research.utoronto.ca/forms/ ) and submitting it to the Local Animal Care Committee. The form is reviewed by the Local Animal Care Committee and then forwarded to the Office of Research Ethics for processing and approval. Hospital-Based Research In the case of a research or teaching project where the funding is administered by the University and the research is to take place at a hospital or related institution, approval for use of animals is obtained through the Animal Care Committee connected with the hospital or related institution University approval is directly obtained by representation of the University on the hospital Animal Care Committee. The Office of Research Ethics requires a complete copy of the current, valid hospital-approved animal protocol, and the assurance that one member of the hospitalbased Animal Care Committee (who may be cross-appointed or affiliated with the University of Toronto) has reviewed and approved the protocol. Detailed Instructions for Filling out the AUPF form These instructions are numbered to correspond with the numbered sections of the AUPF. 1) Fill out personal information in full, ensuring that; Personnel number includes any leading zeroes. The designated alternate faculty contact is not the same as the primary investigator (PI). 2) The title should be concise and succinct; it serves to identify the project for administrative purposes and need not be overly descriptive. All teaching protocols must include the Teaching Protocol Appendix and must also undergo Pedagogical Review (both forms are available from www.research.utoronto.ca/forms/ ). Keywords are used as part of the annual reporting to regulatory agencies. Please select up to six keywords from the dropdown menus that apply to the proposed project. Animal Use Protocol Form (AUPF) Instructions Page 2 of 8
3) The section detailing the lay description of the research requires two pieces of information to be provided. First, the primary objectives of the research project should be described. Secondly, the benefits to human health and scientific knowledge obtained from the experiment should also be described. It is very important that this section be filled out using language appropriate for a reader not familiar with the experimental procedure(s). Many protocols are sent back to the investigator for revisions owing to the technical language used in this section. Use of clear and simple language in this section can potentially save time during the review process. As a guideline, it is suggested that a National Geographic level of writing be used. 4) The categories of invasiveness scale is used to determine the degree of invasiveness of a procedure on an animal. The scale ranges from A-E (see below), with Category A being minimally invasive and Category E causing severe pain or discomfort. Category E studies are not permitted at the University of Toronto. The categories are delineated as follows: Category A: Experiments on most invertebrates or on live isolates Possible examples: the use of tissue culture and tissues obtained at necropsy or from the slaughterhouse; the use of eggs, protozoa or other single-celled organisms; experiments involving containment, incision or other invasive procedures on metazoa. Category B: Experiments which cause little or no discomfort or stress Possible examples: domestic flocks or herds being maintained in simulated or actual commercial production management systems; the short-term and skillful restraint of animals for purposes of observation or physical examination; blood sampling; injection of material in amounts that will not cause adverse reactions by the following routes: intravenous, subcutaneous, intramuscular, intraperitoneal, or oral, but not intrathoracic or intracardiac (Category C); acute non-survival studies in which the animals are completely anesthetized and do not regain consciousness; approved methods of euthanasia following rapid unconsciousness, such as anesthetic overdose, or decapitation preceded by sedation or light anesthesia; short periods of food and/or water deprivation equivalent to periods of abstinence in nature. Animal Use Protocol Form (AUPF) Instructions Page 3 of 8
Category C: Experiments which cause minor stress or pain of short duration Possible examples: cannulation or catheterization of blood vessels or body cavities under anesthesia; minor surgical procedures under anesthesia, such as biopsies, laparoscopy; short periods of restraint beyond that for simple observation or examination, but consistent with minimal distress; short periods of food and/or water deprivation which exceed periods of abstinence in nature; behavioral experiments on conscious animals that involve short-term, stressful restraint; exposure to non-lethal levels of drugs or chemicals. Such procedures should not cause significant changes in the animal s appearance, in physiological parameters such as respiratory or cardiac rate, or fecal or urinary output, or in social responses. Note: During or after Category C studies, animals must not show selfmutilation, anorexia, dehydration, hyperactivity, increased recumbency or dormancy, increased vocalization, aggressive-defensive behavior or demonstrate social withdrawal and self-isolation. Category D: Experiments which cause moderate to severe distress or discomfort Possible examples: major surgical procedures conducted under general anesthesia, with subsequent recovery; prolonged (several hours or more) periods of physical restraint; induction of behavioral stresses such as maternal deprivation, aggression, predator-prey interactions; procedures which cause severe, persistent or irreversible disruption of sensorimotor organization; the use of Freund s Complete Adjuvant (see CCAC Guidelines on Acceptable Immunological Procedures ). Other examples include induction of anatomical and physiological abnormalities that will result in pain or distress; the exposure of an animal to noxious stimuli from which escape is impossible; the production of radiation sickness; exposure to drugs or chemicals at levels that impair physiological systems. Note: Procedures used in Category D studies should not cause prolonged or severe clinical distress as may be exhibited by a wide range of clinical signs, such as marked abnormalities in behavioral patterns or attitudes, the absence of grooming, dehydration, abnormal vocalization, prolonged anorexia, circulatory collapse, extreme lethargy or disinclination to move, and clinical signs of severe or advanced local or systemic infection, etc. Animal Use Protocol Form (AUPF) Instructions Page 4 of 8
Category E: Procedures which cause severe pain near, at, or above the pain tolerance threshold of anaesthetized conscious animals This Category of Invasiveness is not necessarily confined to surgical procedures, but may include exposure to noxious stimuli or agents whose effects are unknown; exposure to drugs or chemicals at levels that (may) markedly impair physiological systems and which cause death, severe pain, or extreme distress; completely new biomedical experiments which have a high degree of invasiveness; behavioral studies about which the effects of the degree of distress are not known; use of muscle relaxants or paralytic drugs without anesthetics; burn or trauma infliction on unanaesthetized animals; a euthanasia method not approved by the CCAC; any procedures (e.g., the injection of noxious agents or the induction of severe stress or shock) that will result in pain which approaches the pain tolerance threshold and cannot be relieved by analgesia (e.g., when toxicity testing and experimentally-induced infectious disease studies have death as the endpoint). 5) The type of experiment should be indicated. If the proposed protocol is for teaching purposes, then the teaching checkbox should be ticked. If it is a research project, then the research checkbox should be ticked. If the project is for testing purposes (e.g. efficacy test of a new drug) then the testing checkbox should be ticked. More than one box may be ticked if appropriate. If a project involves surgery, then please indicate whether it s acute or chronic (survival). Likewise, for projects that do not involve surgery, indicate if the study is acute or chronic. More than one descriptor may apply. 6) The proposed start date of the research described in the protocol should be provided. Please note that the start date cannot be prior to the date of submission to the LACC, and the research may not commence until the protocol has been approved in full. If the proposed research is a continuation of a previously approved protocol or the project is expected to continue for more than a year, please indicate ongoing for the relevant start/completion dates. 7) The past history section is intended to indicate that scientific progress was made on prior iteration(s) of the protocol. If the investigator does not have a currently approved protocol which is similar to (or the same as) the protocol being submitted, then the N/A (New Protocol) checkbox should be ticked. Otherwise, the investigator should answer the two questions pertaining to the recently/currently valid protocol and provide a brief summary of the study s scientific progress to date. Animal Use Protocol Form (AUPF) Instructions Page 5 of 8
8) A detailed description of the proposed project and procedures should be included. Copies of complete grant applications will not be accepted. If the space provided on the AUPF form is insufficient, then additional pages may be appended. Diagrams, flowcharts and tables should be included if appropriate. Clear and concise descriptions which are easily understandable to those with a general science background are desirable. If the members of the LACC cannot understand a procedure or experimental protocol, it may require additional submissions to the committee which may delay the review process. 9) The endpoints section should describe the necessary measures that will be taken in the event of an experimentally-induced negative impact on the health or welfare of the animal (i.e. a disease or life-threatening condition). Appropriate measures include careful ongoing monitoring and consultation with veterinary staff for appropriate treatment or euthanasia should there be any negative impact on animal health. Animals should be monitored by trained laboratory staff, generally on a daily basis. This monitoring should be in addition to any monitoring that is done by vivarium staff. Laboratory staff responsible for monitoring should be specifically named. 10) The University publishes a list of Standard Operating Procedures (SOPs) that govern the performance of many tasks and procedures. All procedures must be performed in accordance with an approved SOP. A list of commonly used SOPs can be found with the Animal Use Protocol Form (see www.research.utoronto.ca/forms/aupf/ ). Please indicate the SOPs that will be used in conjunction with the protocol. It is not necessary to append the SOPs to the protocol if they are listed on the protocol form. 11) All research staff who are working with the live animals should be listed, along with their academic rank or designation. All staff are required to take the University s Short Course on Animal Care (see http://www.medresearch.utoronto.ca/dcm_short_course.html ) which is presented by the Division of Comparative Medicine. Any additions or deletions of research staff must be communicated to the respective Local Animal Care Committee. Additions of research staff will require submission and approval of an AUPF Amendment Form (available from www.research.utoronto.ca/forms/ ) by the LACC. 12) The animals to be used for the research project should be listed in this section. A different protocol must be filled out for each species used (i.e. rats, mice, etc). Different breeds or strains of a single species can be covered by a single protocol, provided they are used for the same study. Information must be provided regarding the source(s) of the animals (e.g. the number of animals that are obtained from suppliers, the number that are to be bred inhouse and the number that are obtained through other means). If multiple breeds or strains of animals are being used, then total numbers should be indicated for each of the three sourcing options. Animal Use Protocol Form (AUPF) Instructions Page 6 of 8
13) The justification section requires that the type of animal and the numbers requested be justified for scientific merit. The choice of species for the study and the rationale for the number of animals requested should be fully explained. Cost is never an acceptable justification for using a given species. When justifying the number of animals to be used, it can be helpful to include statistical calculations or power analyses. 14) Consideration of the 3 Rs (replacement, reduction and refinement) should be undertaken before a study using animals is proposed. The CCAC publishes an excellent online resource at http://3rs.ccac.ca dedicated to the 3 Rs, which has many helpful links to alternatives that can remove or reduce the need to use animals. For assistance in performing a 3Rs search, see the CCAC s 3Rs Search Guide at http://3rs.ccac.ca/en/searches-and-animal-index/guide/. Please complete all three parts of this section. 15) All drugs used for anaesthesia and analgesia should be indicated. The name of the drugs used, their dosages (expressed in mg/kg as appropriate) and the route of administration (e.g. intraperitoneally, IP; intramuscularly, IM; subcutaneously, SC; inhalation) should be provided. Please consult veterinary staff if you are unsure of the proper dosages. Appropriate procedures for monitoring the animals while under anaesthesia and during recovery should be followed, as well as proper intra-operative and post-operative care (see www.research.utoronto.ca/forms/aupf/ for standard operating procedures). Please note that the frequency (number of doses per day) and duration (number of days) of analgesia administration must be indicated. 16) The use of any hazardous agents ( see the AUPF page for more details: http://www.research.utoronto.ca/forms/aupf/ ) if the study requires that they be detailed in this section. Any biological agents (e.g. viruses), chemical agents, carcinogens (e.g. N- ethyl-n-nitrosurea (ENU)) or radioisotopes must be listed. If radioisotopes are to be used, then the radiation permit number and its expiry date must be provided. For the hazardous agents listed, specific details regarding the amounts, routes and half-life must be provided in the relevant section. Any necessary precautions to be taken to prevent harm to humans and/or animals outside the study subjects must be provided. Protocols will be sent back to the researcher for revisions if this section is not completed in full. 17) The provision of euthanasia upon termination of the study must be described. The method of chemical or physical euthanasia should be provided, and strong justification should be given in the event that a physical method of euthanasia will be performed without the prior use of a general anaesthetic. In the event that the animals are not euthanized, the disposition of the animals should be provided (i.e. released back to the wild for certain field studies). Animal Use Protocol Form (AUPF) Instructions Page 7 of 8
18) Please indicate the source of funding. Sources can be external (e.g. NSERC, CIHR, Canadian Diabetes Association) or internal (e.g. faculty/departmental funding, start-up funding). In either case, please indicate whether the grant has been awarded or whether it is still pending. University of Toronto and CCAC regulations stipulate that all research must be peerreviewed for scientific merit. Please see http://www.research.utoronto.ca/faculty-andstaff/research-ethics-and-protections/animals-in-research/ for more information on peerreview requirements. Studies funded by major external granting agencies are considered to have undergone successful peer review during the granting process. Studies that are either not peer-reviewed as part of the granting process, or are internally funded studies, require peer review for scientific merit before the LACC can approve the protocol. Please contact the Office of Research Ethics for further information. 19) Studies that involve field work or trapping of wild animals require that the Field Research Appendix be filled out (available from http://www.research.utoronto.ca/forms/ ). Permit information for the collection of wild animals should be indicated, along with the name of the relevant authority that granted the permit. Detailed information regarding the method of capture, containment methods and transportation of animals from the field, potential capture of secondary/non-target species and any potential injuries that may arise during the capture process should be provided in the relevant sections. Upon completion of the study, the provision for disposal or release of the animals should be described. More Information Please direct any questions regarding the AUPF that are not covered in this guide to Rhain Louis at rhain.louis@utoronto.ca. Animal Use Protocol Form (AUPF) Instructions Page 8 of 8