NUS Institutional Animal Care and Use Committee 000/16

Date revised: Jan 2016

Form expiry date: 30Jun2016

Procedure B

Detailsof Procedures and Management of Pain and Distress

1. Details of Procedures

1.1 Administration of chemicalsubstances(including biochemical, drug, DNA and RNA); Pharmacologic /Toxicologic studies(other than immunisation).

Procedure B

NUS Institutional Animal Care and Use Committee 000/16

Note: Please read the IACUCGuidelines on the Use of Non-pharmaceutical-grade Substances on Animals

Procedure B

NUS Institutional Animal Care and Use Committee 000/16

Please note that all substances should be listed in the Risk Assessment Section B submitted to/approved by OSHE.

(a)Information on the chemical substanceand formulation for administration

Name of substance
(chemicals, drug, biochemicals, DNA and RNA) / Grade of chemical
(provide purity if non-pharmaceutical grade) / Source
(Name of supplier if commercial) / Name of Vehicle, diluents or any excipient
(if used) / Grade of vehicle, diluents or excipient
(provide purity if non-pharmaceutical grade) / Physical state of formulation
(e.g., solution, suspension)

(b)Please explain the selection of the grade of the substance (e.g. pharmaceutical grade, analytical grade, etc.):

Pharmaceuticalgrade. Concentration, formulation and chemical properties are appropriate for the species and route(s) of administration.

Non-pharmaceutical grade because the pharmaceutical-grade compound is not available in the appropriate concentration or formulation or the appropriate vehicle control is unavailable.

Non-pharmaceutical grade because the compound is required to generate data that are part of an ongoing study or that are comparable to previous work.

Non-pharmaceutical grade becausethe chemical properties of the compound and the route of administration are appropriate for the study (e.g., the purity, grade, stability in and out of solution, solution vehicle properties, pH, osmolality, and compatibility of the solvent and other components of final preparation). In some cases the reagent-grade of the compound may be as or more pure than the pharmaceutical-grade.

Other. Please explain the scientific justification for the selection of the grade of the substance:

N.A. The chemical administered is an unadulterated pharmaceutical grade formulation administered to an approved species by an approved route according to the instructions on the product insert.

(i) Sterility– describe how the preparation is sterilized:

(ii) pH –Is the pH of the preparation within the acceptable range (e.g., 7.3 -7.45 for i.m and s.c, 4.5 -8.0 for i.v and i.p)?

Yes.

No. Explain how the effect of pH is minimised:

(iii) Osmolality –Is the preparation isotonic?

Yes.

No. Explain how the effect of non-isotonic substance is minimised:

(iv) Stability – Is the stability of the formulation known?

Yes. The preparation will be used before expiry.

No. Explain how the integrity of the formulation may be maintained:

(v) Are there any known effects (e.g., from the literature, pharmacokinetics studies, etc) on animals due to (i) toxicity, (ii) pyrogenicity and (iii) compatibility of the substances?

Yes. Please address the effect(s) in Section 2 (2.4 a).

No.

(d)Method of administration

(i) Describe the method of administration

By surgical procedure as described in Procedure A (for administration using implant and physical device such as osmotic pump, catheter, etc)

As described below: (include route and size of needle if by injection).

(ii) Complete the table below:

Name of substances: (chemical, drug biochemical, DNA and RNA) / Location of procedure(Bldg, Room No) / Dosage
(amount per unit body weight) / Volume of admin.
(volume per unit body weight) / Route of admin.
(if i.v., indicate which vein) / Frequency of admin. per animal
(how often will the substance be administered) / Total number of admin.per animal

(e) Identification of hazardous substances(i.e., hazardous to animals and to human, such as animal handler and care taker).

Non-hazardous. Indicate which of the above substances are not hazardous and explain or attach supporting documents (e.g. MSDS).

Hazardous. Indicate which of the above substancesare hazardous and complete Procedure D

1.2Administrationphysical devices and implants.

(a)Method of administration

Surgical Method as described in Procedure A

Non-surgical method as described below:

(b)Sources of the device / implant.

(i) Name of supplier of each device / implant if commercially available:

Commercial. Name of supplier:

Home-made

(c)Is the device/implant acceptable for human and/or animal use? (i.e. FDA, Medicines and Healthcare products Regulatory Agency approved, British Standards institution certified or any other accreditation body)?

Yes

No. Please provide the following information:

(1) List the non-pharmaceutical-gradedevice / implant:

(2) Explain why non-pharmaceutical-grade is used:

(3) Described the biocompatibility of the device / implant:

(4) Described how the device / implant is sterilized:

(c) Identification of hazardoussubstances (i.e., hazardous to animals and to human, such as animal handler and care taker):

Non-hazardous. Indicate which device / implant is not hazardous and explain or attach supporting documents (e.g. MSDS).

Hazardous. Indicate which device / implant is hazardous and complete Procedure D

1.3Administration of human / animal blood, body fluids, normal/neoplastic tissues/cells:

(a) Describe the method of administration. If surgery is required for implantation of tissue, the method should be described in Procedure A,but the tissue administered should be listed in the table below.

Method described in Procedure A.

Method as described below:

(b)Please provide the following information and complete Procedure D:

Name of cells, tissues or fluids AND speciesof origin. / Location of Procedure(Bldg, Room No) / Dose
(number of cells) / Volume of admin. / Route of admin. / Frequency of admin.
(how often will the substance be administered) / Total number of admin.

(ii) Please describe how you would ensure that the cells/tissues are pathogen-free.

1.4Administration of biological agents (bacteria, viruses, parasites, prions, including genetically modified agents):

Procedure B

NUS Institutional Animal Care and Use Committee 000/16

Please read the recent multi-agency joint circular onBiosafety Guidelines for Research, Release and Importation of Genetically Modified Organismsand ensure compliance with the guidelines when using genetically modified organisms.

Procedure B

NUS Institutional Animal Care and Use Committee 000/16

(a) Describe the method of administration:

(b)Please provide the following information and complete Procedure D:

Name of agents / Location of Procedure (Bldg, Room No) / Dose
(number of agents) / Volume of admin. / Route of admin. / Frequency of admin.
(how often will the agent be administered) / Total number of admin.

1.5 Collection of blood / fluid (other than ascites) / tissuefrom live animals.

Note: Collection of tissues after animals are euthanized need not be provided. However, collection of tissues from deeply anaesthetised animals (e.g., cardiac puncture for blood collection) needs to be provided even if it is a terminal procedure.

(a)Describe the method of collection:

(b)Complete the table below:

Blood / fluid / tissue to be collected / Location of Procedure(Bldg, Room No) / Route of collection / Volume for each collection / Frequency of collection

1.6 Immunisation / antibody production.

Location of procedure (Bldg, Room No):

(a) Describe the method of immunisation and list the antigen(s) and the adjuvant(s) for initial and subsequent immunisation. If the antigens are hazardous, please submit Procedure D:

Method :
Antigen(s) and adjuvant(s) for Initial immunisation :
Antigen(s) and adjuvant(s) for subsequentimmunisation :

(b)Are the antigen(s) and adjuvant(s) of pharmaceutical grade? (i.e., they are approved for human and veterinary use)?

Yes

No. Please provide the following information:

(1) List the non-pharmaceutical antigen(s) and adjuvant(s):

(2) Explain why non-pharmaceutical grade is used:

(3) Described how they are sterilized:

(c)List or describe injection.List anatomic site, volume administered per injection site total volume administered at one time, injection frequency and total number of administrations. Note: maximum 0.05 ml CFA per site for mice and 0.1 ml per site for rabbits:

Site AND route / Number of sites / Volume per site / Total volume per admin / Injection frequency / Total No. of admin.

1.7 Dietary manipulations or food/fluid restriction.Describe the manipulations and/or restriction including the duration:

Note: Provide scientific justification if (1) neonates are fasted beyond 3 hours, (2) animals are provided with less than normal (ad libitum) food or drinking water, (3) food or fluids are withheld for more than 18 hours.

All animals under dietary manipulations and/or restriction should be monitored frequently for body weight loss (e.g., weighing animals 3 times a week). Proper record of animal weight should be maintained and animals must be euthanized when they have lost 20% of the body weight.

Location of procedure (Bldg, Room No):

1.8 Behavioural studies. Describe the behavioural tests or procedures (including any training of animals prior to the test)and the duration of the test:

Location of procedure (Bldg, Room No):

1.9 Physical restraint studies. Describethe followingif conscious animals will be restrained for more than 15 minutes. To minimize distress or pain associated with restraint,the period of restraint should be no longer than required to achieve the aims of the project.

Location of procedure (Bldg, Room No):

(a) Describe the procedure and the restraint device used to restrain the animals:

(b) Provide and justify the duration and frequency of restraint, and the observation intervals of restrained animals:

(c) Describe the training of animals to adapt to restraint. Please note that animals that fail to adapt should not be used for the study:

1.10 Imaging of live animals. Please complete (a)-(g) below:

Location of procedure (Bldg, Room No):

(a) Describe the imaging modality/procedure to be used (e.g., X-ray, CT Scan, Fluoroscopy, PET scan, MRI, IVIS, Ultrasound etc.), indicate which anatomical part of the animals to be imaged and why this imaging procedure(s) is required.

(b) Please provide details of the administration of imaging probe to be injected in-vivo including imaging contrast agents, fluorescent, bioluminescent imaging reagents and radiation probes (Note: The chemicals should be listed in Section 1.1). The details should include

(i) timing, dose and specific requirements for imaging probe.

(ii) injection method, e.g. using catheters, inhalant delivery, or other methods.

(i) the type of anaesthesia (e.g. induction, maintenance, ventilation – frequency and volume) or restraining devices to be used,

(ii) skin preparation/ hair removal techniques,

(iii) maximum duration under anesethesia/imaging per day, and

(iv) frequency and interval of imaging.

(d) Please specify if there are fasting, diet, light and environmental controls required to alter physiological conditions (e.g. if fasting is required–how many hours before imaging?)

(e) Describe how animals will be monitored during the imaging. Discuss how animals will be kept warm and hydrated if appropriate.

(f) Will blood sampling be required? (e.g. for plasma glucose concentration, metabolite analysis and probe concentration activity over time). If so, please indicate frequency and volume for each sample.

(g) Please identify (i) who will perform the data analysis, and (ii) team member(s) to be registered as a radiation worker and describe their experience with the stated irradiating apparatus/ handling of radioactive materials, if any.

Note: Individuals working with ionising radiation, operating irradiating apparatus/handle radioactive materials/animals must be registered as a radiation worker (i.e., with a valid R1 license).

1.11Other non-surgical procedures. Describe all other non-surgical procedures that will cause more than momentary pain and distress to the animals:

Location of procedure (Bldg, Room No):

2. Management of pain and distress

2.1Will any of the procedures you have described in 1.1 - 1.11above cause more than momentary pain and/or distress?

Yes. (Please describe the nature of pain and distress and how you will manage them in 2.4 below)

No. Please explain:

2.2Will the substance/device or cells administered in 1.1 – 1.4 and 1.6 above cause effects that will result in pain and/or distress at any time post-administration?

Note: The answer should be ‘Yes’ for substances with known toxicity, pyrogenicity, incompatibility or side effects.

N.A. No substance is administered to animals.

Yes.(Please describe the nature of pain and distress and how you will manage them in 2.4 below)

No. Please explain:

Procedure B

NUS Institutional Animal Care and Use Committee 000/16

2.3Some procedures are known to cause serious complications that will result in pain and/or distress if they are not carried out properly(e.g., improperoralgavageor intra-peritoneal injection may cause complications), isthis applicable to any of the procedures you have described in 1.1 -1.11above?

Procedure B

NUS Institutional Animal Care and Use Committee 000/16

Yes. (Please describe the nature of pain and distress and how you will manage them in 2.4 below)

No. Please explain:

2.4If you have indicated “Yes” for any of the questions 2.1 - 2.3 above, describe how you would manage the pain and/or distress as follows:

N.A.

(a) Indicate the procedure and the nature of the pain and/or distress.

Note: For administration of substances, the effects of compound and/or vehicle including intended effects and side effects should be described.

Procedure / Nature of pain and distress
Admin. of substances Admin. of cells/tissuesAdmin. of infectious agents Collection of tissues/ bloodImmunisation Dietary manipulations Behavioural studiesPhysical restraint Other procedure above
Admin. of substances Admin. of cells/tissuesAdmin. of infectious agents Collection of tissues/ bloodImmunisation Dietary manipulations Behavioural studiesPhysical restraint Other procedure above
Admin. of substances Admin. of cells/tissuesAdmin. of infectious agents Collection of tissues/ bloodImmunisation Dietary manipulations Behavioural studiesPhysical restraint Other procedure above
Admin. of substances Admin. of cells/tissuesAdmin. of infectious agents Collection of tissues/ bloodImmunisation Dietary manipulations Behavioural studiesPhysical restraint Other procedure above
Admin. of substances Admin. of cells/tissuesAdmin. of infectious agents Collection of tissues/ bloodImmunisation Dietary manipulations Behavioural studiesPhysical restraint Other procedure above

(b) What criteria will you use to monitor this pain and/or distress?

As described in 8.4 (b) / 3.2 (a) in Appendix 3 of amendment.

As described below, if different from or additional to 8.4 (b) / 3.2 (a) in Appendix 3 of amendment:

(d) What measures will be taken to manage the pain and/or distress?

2.5. If anaesthetic/analgesic is used for any of the procedures and/or for the restraint of animals, please complete the table below:

N.A.

As in Procedure A.

As described below:

Procedure requiring anaesthetic/analgesic / Agent / Dose and volume / Route of administration
Admin. of substances Admin. of cells/tissuesAdmin. of infectious agents Collection of tissues/ bloodImmunisation Dietary manipulations Behavioural studiesPhysical restraint Other procedure above / I.M.ImmersionInhalationI.P.I.V.OralTopicalS.C
Admin. of substances Admin. of cells/tissuesAdmin. of infectious agents Collection of tissues/ bloodImmunisation Dietary manipulations Behavioural studiesPhysical restraint Other procedure above / I.M.ImmersionInhalationI.P.I.V.OralTopicalS.C
Admin. of substances Admin. of cells/tissuesAdmin. of infectious agents Collection of tissues/ bloodImmunisation Dietary manipulations Behavioural studiesPhysical restraint Other procedure above / I.M.ImmersionInhalationI.P.I.V.OralTopicalS.C
Admin. of substances Admin. of cells/tissuesAdmin. of infectious agents Collection of tissues/ bloodImmunisation Dietary manipulations Behavioural studiesPhysical restraint Other procedure above / I.M.ImmersionInhalationI.P.I.V.OralTopicalS.C

-- End of Procedure B–

Procedure B