Animal Use Protocol (AUP) (Application form Instructions) 1. Please refer to the NTU-IACUC Policies for Special Procedures before submitting this application. 2. In order to perform its obligations in accordance with the NTU-IACUC Policies for Special Procedures, it needs a complete description of the proposed animal use and care. Incomplete application will be returned to the applicant resulting in delay in the granting of approval. 3. Submit the completed form together with the soft copy of required documents. 4. Any changes to the approved Animal Use Protocol (AUP) (procedure, species, personnel, etc.) must be documented through submission of an Amendment Form and approved by the NTU-IACUC before implementation. a) Any SIGNIFICANT changes 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) require IACUC review (and approval) prior to initiation. b) Any NON-SIGNIFICANT changes to an approved project in progress (e.g. increase in number of animals used within 15%, changes in location of experiment or changes in personnel) require administrative review (and approval) prior to initiation. 5. Approval of AUP will be for a maximum of 3 years. Extension can only be granted one time for a maximum of 6 months with the approval of the NTU-IACUC. Following this, a new application is required. 6. For animal deliveries please ensure Facilities Director is informed. 7. NTU-IACUC reviews applications monthly so please ensure that this completed form, along with necessary supporting documents, are submitted by the first working day of a month so as to be reviewed the following month. All reviews by NTU-IACUC are on a first come, first served basis. 8. Manuscripts submitted to journals that are based on work done on an approved AUP must include reference to the approving institute and corresponding AUP number. Version: V06 Effective Date: 23 Aug 2018 Document Number: NTU-IACUC-SOP-1.1-F01 Document Name: NTU-IACUC AUP Application Form 1
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Animal Use Protocol (AUP) (Application form Instructions)
1. Please refer to the NTU-IACUC Policies for Special Procedures before submitting this application.
2. In order to perform its obligations in accordance with the NTU-IACUC Policies for Special Procedures, it needsa complete description of the proposed animal use and care. Incomplete application will be returned to the applicant resulting in delay in the granting of approval.
3. Submit the completed form together with the soft copy of required documents.
4. Any changes to the approved Animal Use Protocol (AUP) (procedure, species, personnel, etc.) must bedocumented through submission of an Amendment Form and approved by the NTU-IACUC before implementation.
a) Any SIGNIFICANT changes to an approved project in progress (e.g. the inclusion of new procedures involvingpotential pain, surgery or anaesthetization, or a change in Principal Investigator or course director) require IACUC review (and approval) prior to initiation.
b) Any NON-SIGNIFICANT changes to an approved project in progress (e.g. increase in number of animals usedwithin 15%, changes in location of experiment or changes in personnel) require administrative review (and approval) prior to initiation.
5. Approval of AUP will be for a maximum of 3 years. Extension can only be granted one time for a maximum of6 months with the approval of the NTU-IACUC. Following this, a new application is required.
6. For animal deliveries please ensure Facilities Director is informed.
7. NTU-IACUC reviews applications monthly so please ensure that this completed form, along with necessarysupporting documents, are submitted by the first working day of a month so as to be reviewed the following month. All reviews by NTU-IACUC are on a first come, first served basis.
8. Manuscripts submitted to journals that are based on work done on an approved AUP must include referenceto the approving institute and corresponding AUP number.
Version: V06Effective Date: 23 Aug 2018Document Number: NTU-IACUC-SOP-1.1-F01Document Name: NTU-IACUC AUP Application Form
If user(s) to be added is/are not available in selection lists, please provide the following details of user(s) to be added:
Provide the names of all personnel who will oversee or conduct work under this protocol. Click on "Edit selection" to add users to each category.
2a. Principal Investigator (PI)/Course Instructor
PI / Course Instructor here refers to the main person responsible for the care and use of animals in this
protocol. The PI must be the grant holder.
Last name First name
2b. Emergency Contact(s)
Animal facility staff will try to contact personnel listed in this section whenever an animal is found in moribund condition, in pain or distress. However, NTU-IACUC is not responsible for any animals that, in the professional opinion of the Attending Veterinarian, are suffering in excess of the pain and distress limits approved for this protocol, and which are subsequently euthanized for humane reasons to alleviate further suffering.
In the event that animals have to be euthanized by the Attending Veterinarian without the knowledge of the PI / emergency contact personnel, indicate on cage cards whether the Attending Veterinarian and his staff are to assist in collecting any of the following tissue samples:
a) Type of tissue samples to be collected
b) Harvesting procedures
c) Storage conditions of harvested tissue samples
2c. Co-Investigator(s)
List the names of all other individuals (besides the PI) authorised to conduct procedures involving animals under this protocol.
Last name First name
2d. Research Personnel
Research Personnel conduct research procedures using animals under this protocol.
2e. Pre-Reviewer
Pre-Reviewers are required to perform a pre-review of the protocol as assigned by the PI. The Pre-Reviewer is the ACR or ADR of the School or College respectively.
First name
2f. Request Addition of a New User
First Name Last Name e-mail Contact N0
Last name
First name Last name
First name Last name
Version: V06Effective Date: 23 Aug 2018Document Number: NTU-IACUC-SOP-1.1-F01Document Name: NTU-IACUC AUP Application Form
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Grant Type
Grant Title
Others
Funding Status
Awarded
Period of grant validity (DD-MMM-YYYY):
Pending
Others
Funding
Awarded Date Expiry Date
Version: V06Effective Date: 23 Aug 2018Document Number: NTU-IACUC-SOP-1.1-F01Document Name: NTU-IACUC AUP Application Form
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Species
4a. Animal Characteristics
Age at Acquisition
Weight at Acquisition (metric)
Number Housed at Any One Time
Strain
4b. Animal Source (Local/Overseas, Address)
4c. Housing Location
4d. Other Descriptions (State any other information relevant to animals allocated)
4e. Breeding Plan (Determine and describe the number of breeding pairs you need to supply sufficient mice for your experiments)
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(i
(i) Specific Objectives of the Project
(ii) Project Abstract
(iii) Attach a Flowchart of Experimental Procedures and Timelines * (*)
I have attached a flowchart
No flowchart attached
v) Significance of Expected Benefits to Humans or Animals
Lay Description
List in point form
250 words or less, using simple language, avoiding the use of scientific/medical jargon
The flowchart should give a clear overview of the experimental plan for the animals from entry into the project to proposal's endpoint, with timelines clearly indicated.
Version: V06Effective Date: 23 Aug 2018Document Number: NTU-IACUC-SOP-1.1-F01Document Name: NTU-IACUC AUP Application Form
Use one of the following web-sites to assist / justify your searches
1. National Centre for the Replacement, Refinement and Reduction of Animals in Research
2. USDA Animal Welfare Information Centre, Alternatives and Searches
3. Alternatives to Animal Testing on the Web
4. Johns Hopkins University Centre for Alternatives to Animal Testing
5. European Union Reference Laboratory for Alternative to Animal Testing
6. Fund for the Replacement of Animals in Medical Research (FRAME)
The IACUC requires “that animals should be used only if the researcher’s best efforts to find an alternative have failed”. The three Rs (Replacement, Reduction and Refinement) are the cornerstone of ethical animal research, and NTU-IACUC requires investigators to implement the 3Rs when they are preparing to use animals for scientific or teaching purposes.
6a. Describe the results of searches conducted to verify that the proposed work is not an unnecessary duplication of previous work done.
Search Data
Add literature search information.
Search Searched Keywords Searched to DB Title # of hits conducted from searched
Based on search results obtained, how will the experiments outlined in this protocol be tailored to avoid duplication of earlier work done?
6b. Alternative non-animal methods
Are there any alternative non-animal methods used by other investigators for the type of work proposed in this AUP (e.g. tissue cultures, in vitro monoclonal antibody, computer model, etc)?
Yes
No
6c. Why must animals be used in these experiments?
Select all that apply
This is a study of animal behaviour
This phenomenon under study cannot be reproduced in vitro
This is a pre-clinical study of the in vivo effectiveness of a treatment or procedure
This is for teaching
Others
Describe why these alternatives are not appropriate for this project.
Please elaborate:
IMPORTANT: Retain all search data on file for the duration of the AUP
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6d. Species Characteristics
What characteristics of this/these species make them appropriate for the proposed study? *
These might include structural, behavioural, physiological, biochemical, or other features or considerations (such as availability of species-specific reagents, or the use of well-established model) which make the model compatible with the research objectives. Cost is not a primary consideration.
6e. Reduction, replacement and refinement: How have you considered the 3Rs?
(i) Reduction: Describe how you have attempted to reduce the number of animals. *
(ii) Replacement: Have you considered statistical models in vitro work, instead of using animals etc.? Please describe. *
(iii) Refinement: Have you considered pilot studies, how have you refined any surgical procedures or other manipulations? Please describe. *
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NO pain, distress, or use of pain-relieving drugs. Routine procedures (e.g. injections, tattooing, blood sampling) should be reported with this classification.
Species Strain YEAR 1 YEAR 2 YEAR 3 Remarks
Accompanying pain or distress to the animals for which appropriate anaesthetics, analgesics, or tranquillising drugs will be used.
Species Strain YEAR 1 YEAR 2 YEAR 3 Remarks
Accompanying pain or distress to the animals for which the use of appropriate anaesthetics, analgesic, or tranquillising drugs would have adversely affected the procedures, results, or interpretation of the research, experiments, surgery
Species Strain YEAR 1 YEAR 2 YEAR 3 Remarks
Animals being bred, conditioned, or held for scientific activities at each facility but not yet used for research purposes.
Species Strain YEAR 1 YEAR 2 YEAR 3 Remarks
Complete the table below to indicate the total number of animals requested per year. This should be a sum of the annual number requested under the selected pain categories:
Species Strain YEAR 1 YEAR 2 YEAR 3 Remarks
Version: V06Effective Date: 23 Aug 2018Document Number: NTU-IACUC-SOP-1.1-F01Document Name: NTU-IACUC AUP Application Form
Explain the procedures producing pain or distress in these animals and justify why appropriate anaesthetic, analgesic or tranquilising drugs cannot be used.
*
If pregnant animals are used, describe how you will take care of the offspring. *
7c. Please justify the number of animals used as stated in section 7.a.
Provide a breakdown of how the total numbers are arrived at and explain the distribution of animal numbers across different procedures or experimental groups. *
If statistics are used (e.g. c2 test, Fisher’s Exact test, Student’s t-test, ANOVA), describe the statistical analysis.
7d. Use of pregnant animals
Version: V06Effective Date: 23 Aug 2018Document Number: NTU-IACUC-SOP-1.1-F01Document Name: NTU-IACUC AUP Application Form
In the field "Name of study segment", enter the AUP title. Expand the "Species" field and select all the species reflected under this application and click "Apply". Lastly, select "Add".
Version: V06 Effective Date: 23 Aug 2018 Document Number: NTU-IACUC-SOP-1.1-F01 Document Name: NTU-IACUC AUP Application Form
Will food/fluid or both be restricted for the animal(s) used?
Yes *
Provide additional details, including (i) duration of restriction, (ii) possible effects on the animal, (iii) how the animal’s health and well-being will be assessed during this period and (iv) why positive reinforcement is not a suitable substitute:
No
Will the animal be restrained conscious for a continuous period exceeding 15 minutes?
Yes *
Provide additional details, including (i) description of the restraint, (ii) duration of restraint and (iii) how the animal will be observed during this period.
If the duration exceeds 6 hours and the animal’s ability to access food and water is limited, provide the following information:
No
Click on "Add/edit procedures" below to select from a list of pre-determined non-surgical procedures. Each procedure will correlate to a training competency under the "Procedures Training Verification" module. If a non-surgical procedure is not listed, manually describe your procedure; note there is no automatic link to training and you will need to document additional training details under the "Procedures Training Verification" module.
List of Non-Surgical Procedures
Description Species
Description of Non-Surgical Procedures
In LAY LANGUAGE, describe each procedure listed above. The following must be included clearly for each procedure, where applicable: Liquids - Volume, Concentration, Route of Administration, Frequency Implants - Dimensions, Size
Others
(a) when food and water will be given (b) how body weight will be monitored (c) how hydration status will be monitored
Version: V06 Effective Date: 23 Aug 2018 Document Number: NTU-IACUC-SOP-1.1-F01 Document Name: NTU-IACUC AUP Application Form
Select the relevant training and vaccinations required.
AUP Training
RCULA Theory Certification
RCULA Practical Certification
Status
The selected training and vaccination are required for all personnel prior to protocol submission and approval. The matrix below shows the current status of these requirements for personnel added to this protocol.
Under Status, the availability of certification and vaccination information is reflected. If any item is indicated with a NO, provide relevant document/update to IACUC Secretariat so that the user’s profile can be updated.
Tentative Training and Vaccination Dates
Select the vaccinations applicable to this study
Tetanus (compulsory for work with rodents)
Hepatitis B (compulsory for work with rodents)
Measles (compulsory for work with primates)
All personnel listed must have undergone RCULA training and have the necessary vaccinations. If RCULA training or vaccination are not complete, state tentative completion dates:
RCULA Training
Personnel Tentative training date (dd/mm/yyyy)
Vaccination
Personnel Vaccination Tentative vaccination date (dd/mm/yyyy)
Version: V06Effective Date: 23 Aug 2018Document Number: NTU-IACUC-SOP-1.1-F01Document Name: NTU-IACUC AUP Application Form
Indicate any clinical conditions or abnormalities expected or that could arise as a result of the proposed study or teaching exercise
In terms of species-specific behavioural changes and physiological signs, list the criteria that will be used to trigger the decision to remove an animal from the teaching exercise or experiment, or to terminate the teaching exercise or experiment.
Select all the humane endpoints that apply:
Tumour size
Specify maximum growth (cm):
Weight loss (more than 20% of bodyweight over 1 week OR no more than 10% over 24 hrs)
Inability to drink or eat
Abnormal behavior such as vocalization, CNS signs, shivering, immobility, inactiveness
Vomiting of blood (Haematemesis): NOT applicable to rodents
Severe bleeding
Severe or chronic pain
Others:
e.g. behavioural changes such as increased grooming, vocalization or postural changes, or physical abnormalities such as anorexia, dehydration, diarrhoea, etc.
If necessary, consult the Attending Veterinarian for further advice.
10a. Clinical conditions or abnormalities expected from proposed study
When experimental procedures produce animals that may become ill, it is necessary to define an endpoint to ensure that an experimental animal's discomfort, pain and/or distress is terminated, minimized or reduced.
10b. Criteria for animal removal from study
10c. Humane Endpoints
Version: V06Effective Date: 23 Aug 2018Document Number: NTU-IACUC-SOP-1.1-F01Document Name: NTU-IACUC AUP Application Form
List all genetic constructs for Genetically Engineered Animals used on this protocol.
GMAC approval must be obtained and attached to this section tab
3rd generation lentivirus vector are exempt from BATA. Please select ABSL2 for biosafety level. Please note that a maximum of 100ml of Lentivirus culture may be brought to NTU and that there should be no storage within the facility.
The Singapore Biosafety Guidelines for Research on Genetically Modified Organisms (GMOs) can be accessed for further information: http://www.gmac.gov.sg/Index_Singapore_Biosafety_Guidelines_for_Research_on_GMOs.html Or please contact the GMAC Secretariat: [email protected]
List all Radiological Hazards used on this protocol.
Complete the table below and attach appropriate license(s) for the use of these radioactive substances.
List all Chemical Hazards used on this protocol.
Complete all the relevant sections below if this AUP uses any of the following material(s). Indicate the nature of the material(s) and biological safety levels where appropriate. Prior to the start of the experiment involving the use of potentially hazardous material at NTU, the PI and/or his /her staff must inform the room technician and ARF safety officer the nature of the work and material(s) used. All cages must be labelled with the material used, start date, estimated end date, and contact details of user. All activities within the AUP must be covered by a Risk Assessment (RA) and please provide the approved RA with the application.
1) Potentially Hazardous Materials
A. Genetically Modified Organisms (GMOs)
B. Radioactive Substances
(i) Are radioactive substances used?
No
Yes
Please select an item for each mandatory (*) section and provide a text where necessary.
(i) Are chemical substance(s) used?
Appropriate Safety Data Sheet (SDS) for the chemical(s) used must be included as well as SOP on waste management.
A copy of NTU’s Cytotoxic and ABSL2 Waste Handling SOP can be requested from NTU-LKCMedicine ARF safety officer for this AUP’s
waste management consideration.
Note: ONLY WORKING VOLUME MAY BE BROUGHT INTO THE ANIMAL FACILITY. NO STORAGE OF CHEMICAL IS PERMITTED
WITHIN THE FACILITY.
Yes
Isotope Half Life Maximum dose/volume Carcass waste generation (kg/month)
C. Chemical Substances and New Chemical Entities
GMAC reference number:Proposed category of GMAC works:
Cat. ACat. BCat. C
Version: V06Effective Date: 23 Aug 2018Document Number: NTU-IACUC-SOP-1.1-F01Document Name: NTU-IACUC AUP Application Form
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(b) Novel Compound or NCEs
Comment
Please justify and submit supporting document for alternative Waste Management Procedure for IACUC review and
consideration.
Justification:
Describe waste disposal methodology in the Comment box if different from NTU’s Cytotoxic and ABSL2 Waste Handling SOP. Please
also justify the reason for the different method used.
Chemical Name Risk Group Carcass waste generation (kg/month)
D. Human Derived Materials
New restrictions are in place with regards to the use of human derived materials under the Human Biomedical Research Act. All use of human material in animals will require you to file for IRB approval. IRB approval letter must be included in the AUP application or amendment. Restricted human biomedical research is defined in the Fourth Schedule of HBRA and these include:
1. Human biomedical research involving human eggs or human embryos 2. Human biomedical research involving —
(a) the following types of human-animal combination embryos:
(i) cytoplasmic hybrid embryos;
(ii) human-animal combination embryos created by the incorporation of human stem cells (including induced pluripotent stem cells);
(iii) human-animal combination embryos created in-vitro by using —
◾ human gametes and animal gametes; or ◾ one human pronucleus and one animal pronucleus; ◾ the introduction of human stem cells (including induced pluripotent stem cells) into a prenatal animal foetus or
animal embryo;
◾ the introduction of human pluripotent stem cells (including induced pluripotent stem cells) into a living postnatal animal;
◾ the introduction of human stem cells (including induced pluripotent stem cells) or human neural cells into the brain of a living postnatal animal; or
◾ any entity created as a result of any process referred to in sub paragraphs (b), (c) and (d)
For human-derived materials requiring IRB approval, please contact the Secretariat of the NTU Institutional Review Board (IRB) at (65)
65922495 or email at [email protected]. The IRB approval letter obtained should include the use of human cells in animals.
Research involving the use of commercial cell lines (excluding embryonic and other stem cells) will not require NTU-IRB approval
for exemption. Principal Investigators will only need to complete and submit an E-Declaration form to the NTU-IRB via email at
[email protected]. The E-declaration form is obtainable from NTU-IRB office. Please forward the E-declaration form and acknowledgement
letter from NTU-IRB to NTU-IACUC office for record keeping.
HBRA Regulatory Framework
FAQs on HBRA
A copy of NTU’s Cytotoxic and ABSL2 Waste Handling SOP can be requested from NTU-LKCMedicine ARF safety officer for this AUP’s
waste management consideration.
Please select ABSL2 for biosafety level (See section H – Overall Animal Biosafety Level).
Comment
Version: V06Effective Date: 23 Aug 2018Document Number: NTU-IACUC-SOP-1.1-F01Document Name: NTU-IACUC AUP Application Form
(i) Are Materials Defined as per Items 2a-2e Used?
Yes * IRB Application Number:
* IRB Application Approval Date:
Please list and provide information on all human derived materials (examples includes but not limited to ell lines, human stem cells, tissues, etc.) used in this AUP in the table below:
Name Source of Supply (e.g.: from)
E. Non-Human Derived Materials
Are Other Biologically-Derived Materials Used?
Yes
Complete the table below and attach relevant documents
Name of non-human biologically derived material used Source of Supply (eg,:from)
Carcass waste generation (kg/month)
List all human or animal derived Biologicals or Cell Lines used on this protocol.
Date, late MAP test IBC Protocol # IBC Approval Date Source
(i) Are Biological Agent(s) or Toxin(s) Used?
Yes
Please provide information on biological agent(s) or toxin(s) used in this AUP in the table below:
Name Strain / LD50
(for toxins) Supply Source
Select the schedule in Biological Agents and Toxins Act (BATA), if applicable:
First Schedule Part I
First Schedule Part II
Second Schedule
Third Schedule
Fourth Schedule
Fifth Schedule
Not applicable
No
No
No
F. Biological Agent and/or Toxins(s)
Biological or Cell Line
Version: V06Effective Date: 23 Aug 2018Document Number: NTU-IACUC-SOP-1.1-F01Document Name: NTU-IACUC AUP Application Form
Provide toxicity and disposal information; attach necessary documentation:
Complete the table below:
Provide information on other inorganic or organic materials used:
e.g. microchip implantation, gel matrix implantation, etc.
Name Toxicity Information Source of Supply
No
H. Overall Animal Biosafety Level
Overall Animal Biosafety Level and Relevant Chemical Safety Classification
ABSL1
ABSL2
ABSL3
Other Comments
Only applicable if GMOs / human derived materials / biological agents / toxins are introduced or used in this AUP. Attach a risk assessment for all the procedures mentioned in this application including transfer of animals, if any.
2) Staff Health
It is the responsibility of the Principal Investigator (PI) to inform NTU staff of any risks to staff health.
State whether the procedure(s) will result in the release of infectious or non-infectious organisms, and how you intend to handle spills and unused stocks:
Potential hazard to humans:
None
Present
Describe potential health risks to animals or humans
Have you attached a Risk Assessment? (RA)
Yes
No
Specify any special animal care required because of the hazard(s) involved, precautions to be taken by personnel and any special containment requirements (i.e. storage, waste/disposal requirements, etc).
Access the Workplace Risk Assessment System (WRAS) to complete a RA.
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3) Declaration of Biological Material / Animal Products for Use in Animals
Yes
No
If derived from humans, materials MUST be tested for Mycoplasma. Such tests must be performed before this application is submitted. Contact NTU ARF for assistance.
To prevent the inadvertent infection of research animals and NTU colonies, declare the origin and sterility status of any biological materials/animal products you intend to use in this AUP:
Complete the table below: *
The listed materials are PCR based or MAP/RAP/HAP/Mycoplasma tested:
Nature of Material (e.g. cell lines, antiserum, etc.)
Animal Source Supplied
Sterile? (Yes/No)
Supplied Attenuated? (Yes/No)
Version: V06Effective Date: 23 Aug 2018Document Number: NTU-IACUC-SOP-1.1-F01Document Name: NTU-IACUC AUP Application Form
By submitting this form, as the PI you agree to the following terms:
◦ Animals used in this research or teaching project will be cared for in accordance with the principles contained inGuide for the Care and Use of Laboratory Animal (8th Edition)
◦ You have considered alternative procedures that do not involve the use of living animals.
◦ You will use the minimum number of animals consistent with objectives of described research/teaching program.◦ You have carefully selected the species that you propose to use.◦ You will use techniques and facilities that are in accordance with the Guide for the Care and Use ofLaboratory Animal (8th Edition)
◦ You will notify the IACUC of any revisions to this AUP.◦ You will keep copies of all approved AUPs, revisions and amendments in an accessible file.◦ This project has been reviewed for scientific merit.◦ The consultant Attending Veterinarian has been consulted prior to AUP submission / consultation not considerednecessary.
◦ Animals housed in other Satellite Rooms. Animals from external sources need to be quarantined or housed accordingto the revised guidelines on animal quarantine.
◦ Animals will be housed in Animal Facility◦ Facility management has been consulted and has certified that indicated facility(ies) has(have) the resources tosupport this work
Version: V06Effective Date: 23 Aug 2018Document Number: NTU-IACUC-SOP-1.1-F01Document Name: NTU-IACUC AUP Application Form