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2017

Animal Care and Resource Center Animal-Use Research IAEC Project Proposal

& animal imports Guidelines.

All relevant Forms and documents need to be downloaded from the

ACRC website: https://www.ncbs.res.in/research-facilities/acrc

Internal Animal Users Committee

NCBS AND INSTEM, BENGALURU, INDIA

ACRC - https://www.ncbs.res.in/research-facilities/acrc

1 2017_ NCBS Animal Care Resource Center_IAUC & IAEC project proposal detailed guidelines

Contents

I. Animal related research project proposal, approval and commitment process: .................. 3

II. Mandatory resubmission, review, and IAUC & IAEC approval in any modifications of

the initial project proposal terms ................................................................................................. 9

III. Yearly Progress Report .................................................................................................... 9

IV. The IAEC meeting day requirements ............................................................................... 9

V. Ordering and importing Animals from other facilities / labs ......................................... 10

VI. AF Internal User Fees .................................................................................................... 15

VII. Publication Requirements .............................................................................................. 15



I. Animal related research project proposal, approval and commitment

process:

Prior the use of animals for a given project can be authorized: all research projects involving the

use of vertebrate animals and the use of the NCBS animal facility need to be submitted to and

approved by the IAUC (Internal Animal Users Committee) and to the IAEC (Institutional

Animal Ethics Committee) following strict guidelines and requirements.

All proposals should meet the Governmental requirements decided by the Committee for the

Purpose of Control and Supervision of Experiments on Animals (CPCSEA).

(see http://envfor.nic.in/division/committee-purpose-control-and-supervision-experiments-

animals-cpcsea for further information).

Basically: all Animal-Use research projects should be proposed following these different steps:

A. The CPCSEA “Form B (per rule 8(a)”

The CPCSEA “Form B (per rule 8(a)” needs to be filled and submitted to the IAUC and then to

the IAEC.

- The NCBS or Instem IAEC FormB can be found on the ACRC website:

https://www.ncbs.res.in/research-facilities/acrc

- For external institutes please refer to the CPCSEA form:

http://www.moef.nic.in/sites/default/files/Revised_Form_B.pdf

The “Form B” Proposal should include:

Part A-6): Duration of the project (can NOT exceed THREE years; projects will need to

be submitted for extension at the end of their 3rd year. Please make sure that the dates in

A-6 and B-5 are identical- Projects can only be started once approved by the IAEC.

Part B- 1) project title needs to match the title given on the relevant Institutional

BioSafety Committee (IBSC) project proposal.

Choose project title with care. Extra mural grants agencies (DBT, Wellcome-DBT, DST etc)

require certificates of IAEC approval. For this, the approved proposal title and grant application

title must match. If more than one grant application is to be based on a single IAEC approved

project, include subtitles in IAEC proposal.

http://envfor.nic.in/division/committee-purpose-control-and-supervision-experiments-animals-cpcsea

http://envfor.nic.in/division/committee-purpose-control-and-supervision-experiments-animals-cpcsea


http://www.moef.nic.in/sites/default/files/Revised_Form_B.pdf



Note: Any Hazardous substances or genetically engineered reagent to be applied or injected to

live animals or embryos need to be pre-approved by the IBSC (Institutional BioSafety

Committee – please contact Harleen ([email protected]) for information on the IBSC

proposal deadlines and requirements).

Part B- 2 & 3) List of names of all individuals authorized to conduct procedures under

this proposal. (The list can be updated at the next IAEC project review session if

required)

Please note that only InStem or NCBS PIs or facility heads need to be listed as IAEC project PIs.

For grant purposes: post-docs or students will need to be listed as co-guides if required.

Part B- 4) Source of funding for the project: grant / funds details and documents need to

be provided as well as the Institutional Budget Code to which the Animal Housing fees

are to be charged to.

Part B-5): Duration of the project (can NOT exceed THREE years; projects will need to

be submitted for extension at the end of their 3rd year. Please make sure that the dates in

A-6 and B-5 are identical- Projects will only be started once approved by the IAEC.

PartB-6 & 7) Study plan and Objectives

Please give a short outline of the project mentioning the previous data supporting the use of the

specified animal models and planned surgical procedures or reagents to be applied/injected to the

animals.

Detailed study plan must be given in newspaper language (layman terms), which can be easily

understood by the socially aware nonscientific members of the committee.

Note: if your project involves the use of the NCBS Mouse Genome Engineering Facility

(MGEF) services to generate novel Transgenic, KI, KO cKO mice: you should provide the

justification of using these techniques and services for your project and the corresponding

research study plan involving the use of these animals once the first founders will be delivered to

you.

The NCBS Mouse Genome Engineering Facility can only accept projects once they have been

approved by the IAUC & IAEC and the IBSC.

mailto:[email protected]



Part B-8) Animals Required.

Please detail the number of experimental animals required for each study group per year. Respect

the given IAEC table format indicated the different study group categories in columns 1, 2, 3, 4,

etc. Only the actual number of animals to be sacrificed/used for actual terminal experiments

need to be accounted for here. Do NOT indicate the number of animals required for breeding

and expansion of your colonies- and do NOT account for the number of Non-Required animals

which will be produced as result of the breeding scheme but which will not be used for

experiments because they do not match required experimental genotype/ gender/ fitness criteria.

If requesting the services of MGEF to generate new mouse models: you do NOT to account for

the numbers of donor animals required by the MGEF to generate the new Strains but you should

account for the number of experimental animals resulting from the new strain to be used once the

new model will be obtained.

- Statistical power analysis:

o Justifying the required number of animals in experimental groups is a must.

Include power analysis.

o In cases where power analysis does not make sense, such as mouse genetics

experiments involving mutants etc., justify the required number by a bottom-

up estimation. For example, number of mutants needed per experiments,

number of experiments etc.

Please note: The concern for IAEC is only the total number of experimental ANIMALs – NOT

the required number of housing cages. Allocation of cage space for your lab is User committee’s

(IAUC) concern; for this, breeding and additional details on the different animal and cage

requirements should be given in the <ACRC cage space request Form>.

Due to space limitations, expansion of cage space over 200 cages requires approval from the

IAUC and the deans of NCBS or inStem. Additional cage space above 200 cages will only be

granted on a fixed-term basis, not to exceed 6 months.

- Breeding and additional details on the different animal and cage requirements should

be given in the <ACRC cage space request Form> (to download from the ACRC

website). Table 1 of this cage space request form can be added as an annexure to the

IAEC form B if required-

https://www.ncbs.res.in/research-facilities/acrc-IAEC

https://www.ncbs.res.in/research-facilities/acrc-IAEC



Part B-8f: Source of Animals: please give the laboratory / repository from which the animals will

be imported if not yet housed at NCBS- Otherwise: please give NCBS Animal Facility as

<source of animals>.

Rational for the animal usage

Procedures and surgeries to be used- All procedures / surgeries to be operated on animals

need to be clearly detailed with corresponding measures of anesthesia and live surgery

procedures if applicable. The IAEC will mostly focus on the procedures, surgeries and

drugs or reagents that are going to be applied to animals. All of these aspects should be

detailed ensuring that animal pain and welfare are properly assessed, controlled and

procedures justified. Typical anesthesia used ta NCBS facility are Isoflurane Vaporiser or

eventually Xylazine/Ketamine or Avertine injections but these need to be justified with

our Veterinarians.

Part B- 15) Methods of animal disposal. NCBS can either euthanize animals via C02

overdose, anesthesia overdose (Isoflurane) or cervical dislocation. Carcasses are disposed

in deep-freezers followed by Incineration-

Part B- 16) Details on eventual bio-materials to be injected or applied to the animal

(hazardous agents, radio-nucleotides, drugs, chemicals, pathogens, microorganisms,

recombinant DNA/RNA/proteins or cells / other.

Please NOTE: As mentioned in the IAEC – “formB”:

- All substances, (DNA/RNA/protein/drugs, viruses, chemicals, cells, reagents,

irradiation procedures etc.) to be applied or injected into live animals or live embryos

need to be clearly described. Relevant doses and frequency of administration need to

be detailed and any Hazardous substances or genetically engineered reagent need to

be pre-approved by the IBSC (Institutional BioSafety Committee – please contact

Harleen ([email protected]) for information on the IBSC proposal deadlines and

requirements).

NOTES before sending the FormB for review:

a) Yearly Progress report if required (see chapter III);

b) your updated IBSC certificate approving the procedures/reagents referenced in your

IAEC proposal (the project title mentioned on the IBSC certificate needs to match the

IAEC project title);

c) proof of funding source with relevant budget code.

d) any associated wildlife /CPCSEA permit if required for wildlife/field studies;

e) updated cage space request form if applicable (for mice and rat studies).




f) Please date and sign the Cover page and final page of Form B.

B. Details on ALL animal requirements, necessary breeding schemes and

TOTAL number of CAGES required: the “IAUC_Cage Space Request”

document (downloadable from ACRC website)

In addition to the basic animal requirements asked in “form B”, the IAUC asks for more detailed

information regarding the experimental animal groups needed for the pursuit of the project as

well as a detailed breeding scheme required to meet the experimental needs.

The aim here is for the IAUC and the PI to have a more accurate notion of the number of cages,

the space and timeline requirements for each proposed project as well as the estimated Animal

Facility Use costs involved.

In order to estimate yur final required number of cages per day: you wuld need to estimate the

following:

Details on the required experimental animals

- Species / strain /line /genotype

- Age / weight /or size

- Gender (F/M)

- Specific generation or phylogenic relationship if required (F1, F2 …/ littermate

requirements?)

- Number of animals per experimental group to be used simultaneously and frequency of

usage (daily / weekly / monthly / yearly / other)

- Special housing requirements (VivariumV1, V2, V3 or V4; SPF.1 /SPF.2 / non SPF /

ABSL2 /ABSL3 / desired number of animals (breeding and experimental) per cage -

especially if animals are required in solo cages ( = one animal per cage).

- Details and information on the ACRC different SPF level requirements, pathogen

monitoring results and associated housing fees can be found on the ACRC website:

https://www.ncbs.res.in/research-facilities/acrc-useful-links

https://www.ncbs.res.in/research-facilities/acrc-useful-links



Breeding scheme outline necessary to meet the needs in experimental animal numbers

and specifics

- Breeding type (inbred / outbred / outcrossed / backcrossed / hybrid/ other)?

- Number of Breeding duos (= 1 male + 1 female) / trios (= 1 male + 2 females)?

- Continuous breeding or timed and prescheduled breeding?

All these different breeding requirements will impact the end number of cages required

for breeding and expanding your colony: you should therefore have a clear idea of these

requirements before you start your animal protocol. The ACRC can help you in figuring

out what best meets your experimental needs: please do contact us if required.

The IAUC will offer help in filling the “IAUC_Cage Space Request Form” to any PI who

requests it but at least 1.5 weeks PRIOR the submission deadline.

C. Estimates of the total required number of cages per animal holding room

type per year

Once the IAUC has received the “Form B” (appendix F) and the " IAUC_Cage Space Request

Form”; together with the PI, the committee will conclude on the estimated space and timeline

project requirements based on the following numbers:

Breeding cage numbers / animal holding room type / duration time

Gestation / weaning cage numbers / animal holding room type / duration time

Experimental cage numbers / animal holding room type / duration time

If a projects evolves and the PI/Lab needs additional cage space. This request for additional cage

space can be formulated to the IAUC any time during the year by filling out a revised

“IAUC_Cage Space Request Form”. The IAUC will review the request and decide on the

additional number of cages and vivarium type that can be allocated to the PI.

It is important that PIs go through the IAUC approval before expanding their cage numbers as

the IAUC needs to assure that the space is available in the proper vivaria type and decide in

which room these additional cages can be given.

Labs who exceed their IAUC allocated cage numbers without prior “additional cage space

request” to the IAUC will be fined accordingly (see AF user fees paragraph).



Please NOTE: Due to space limitations, expansion of cage space over 200 cages requires

approval from the IAUC and the deans of NCBS or inStem. Additional cage space above 200

cages will only be granted on a fixed-term basis, not to exceed 6 months.

II. Mandatory resubmission, review, and IAUC & IAEC approval in any

modifications of the initial project proposal terms

The final approved proposal should be strictly respected by the PI and its lab members. In case

the project evolves and requires different or additional animals, new strains, adjusted cage

numbers, additional procedures, new lab members to be added on the project or extension in

duration of the project: the new project requirements should be submitted to and approved by the

IAUC and IAEC as amendments to the initial proposal.

In this case, an updated/modified “project proposal FormB” needs to be submitted by the PI to

the IAUC and IAEC. These proposed project modifications or extension dates request should be

clearly highlighted in different color text and distinguishable from the rest of the original project

proposal content.

III. Yearly Progress Report

- The IAEC approves projects for given period (usually maximum 3 years) and an IAEC yearly

progress report form (downloadable form the ACRC website) needs to be submitted every year

attached with the original approved FormB in order for IAEC to allow the project to be pursued.

- PIs who wish to renew or extend their project beyond the original IAEC approved finished date

need to resubmit a new IAEC-FormB as an extension request before the end date of the project

(see above paragraph).

IV. The IAEC meeting day requirements

The IAUC (INTERNAL Animal Users Committee) meets regularly (up to 2 times a months or

whenever necessary) to discuss different aspects of Animal Facility management related issues,

to review users AF related requests or comments and to pre-review all project proposals (FormB)

before they are submitted to the IAEC.



However the NCBS and InStem IAEC (INSTITUTIONAL Animal Ethics Committee) only

meets twice a year (usually December-January and June-July ) to review and approve the

different submitted FormB-project proposals.

Please note that other Institutes might have different IAEC meeting policies. Each Lab should

refer to their home Institute IAEC policies and requirements.

As mentioned earlier: only Faculty can be associated as project PIs. External collaborators (if

any) co-directing the project have to be listed as Co-PIs / Co-guides.

In the context of the NCBS and InStem IAEC meetings; Students and post-docs cannot be listed

as project PIs and they are not the ones to be sent to present the project at the IAEC project

submission review meeting. PIs or Co-PIs need to be the ones presenting their project in front of

the IAEC following these guidelines:

PIs who have submitted their projects for review (either for extension or modification/addition of

new procedure or as new proposals) are expected to make a short presentation based on the

following guidelines:

i) Presentations must be made by the PI 'in person'.

ii) Time of presentation will be strictly limited to 15mins, which will include 5-10

minutes speaker's time and 5 mins discussion.

iii) Presentations should be made with the aid of 3 slides, including experiment

design/flowchart, statistical analysis to justify numbers of EXPERIMENTAL

animals (not number of cages nor breeding requirements) proposed, and any changes

from previous proposals.

iv) Make sure to include copy of relevant approvals from other statutory committees

(IBSC, etc…) or any associated wildlife /CPCSEA permit if required for

wildlife/field studies.

V. Ordering and importing Animals from other facilities / labs

The NCBS Animal Facility will NOT order or receive animals from any Indian commercial

vendors / collaborators which are not registered under the Indian CPCSEA.



ACRC will guide you in any animal or cryosample import process- Briefly: the procedures are

the following:

Any international animal import need advance import license issued from Directorate General of

Foreign Trade, New delhi (DGFT) and ‘NOC’ from regional quarantine officer. (see paragraph

below for specific import license procedures).

External animal requests and imports must be submitted as part of a specific research

proposal approved by the IAUC and the IAEC.

For all animal imports to the facility, it is made mandatory that PIs submit their Animal use

research project proposal justifying their mice imports before the animals can be shipped to the

facility.

As for all other Animal-use research project proposal, the <Animal Import Request> and

eventual associated <cage space request> forms submitted to the IAUC should include the

specifics Housing, Breeding and Cage requirements details for each newly imported strain.

The cage requirements for “Animal Imports” should be detailed in THREE distinct phases:

- Phase 1: The PI should fill the “Animal Import Request form” detailing the actual

number of animals which are being imported, as well as the gender and age group - how

should the imported animals be segregated in each cage during the quarantine period.

(See the “quarantine procedures” paragraph below for more details – importantly: no

breeding is allowed during quarantine).

- Phase 2: the post-quarantine NON-experimental Animal Housing period - providing the

basic colony maintenance breeding scheme details and cage number requirements for

each of your imported strains once they exit the quarantine and before animals are set up

of colony expansion breeding & experimental breeding needs.

Phase 2 (colony maintenance) breeding information should be provided as a separate

“colony maintenance phase” entry of the "IAUC_Cage Space Request Form discussed

previously.



- Phase 3: actual experimental breeding and cage number requirements as detailed

“experimental” entries of the "IAUC_Cage Space Request Form” discussed previously in

paragraphs I-B &I-C.

If the imported animals are not to be experimentally used immediately after their quarantine exit.

Phase 3 details can be given after the animals are imported. However Phase 1 and Phase 2 details

are required by the IAUC to ensure that the facility has the available cage space to welcome and

maintain the animals upon arrival.

Phase 3 details however will have to be submitted and approved by the Institution before any

colony expansion and experimental cage space can be allocated to the lab for all newly imported

strains.

Providing the Budget Code / funding source to which the import fees will be charged

Who is going to pay for the import fees and the future Animal Housing fees once the animal

arrive at the NCBS Animal Facility? The funding source and/or the Institutional Budget Code to

which these fees will be charged to need to be provided.

Checking for any required MTA approvals and obtaining the correct institutional MTA

approval

It is critical that PIs check the MTA (Material Transfer Agreement) requirements for ALL the

animal strains they wish to use for their research. It should also be verified that the MTAs

obtained for a given animal strain is specific to the Institute (NCBS or InStem or C-Camp) that is

going to use this specific strain.

This is obviously true for all animals that need to be imported from external facilities/labs to the

Animal Facility. But importantly: this may also be true for animals already housed within the

different Animal Facility Vivarium. MTAs are generally bipartite contracts signed by TWO

institutes, labs or repositories and should be looked at carefully.

In case of JAX mice: we have recently obtained an agreement allowing the sharing of JAX mice

between InStem and NCBS labs. But this might not be the case for mice coming from other

repositories.



Publishing data with animals under invalid MTAs conditions can have serious legal

consequences for the PI and for the hosting Institute.

Obtaining research animal import license

The following documents are required for obtaining advance import license:

1. Duly filled “Animal import request form” approved by IAUC (downloadable from the

ACRC website).

2. Proforma invoice (if you are purchasing animals from a commercial vendor) or donor

letter (if you are receiving animals as a gift from collaborator).

3. Approval from IAUC and IAEC.

4. Budget code for obtaining DD towards application fee.

Obtaining the Specific Pathogen monitoring Veterinarian certificate from the lab of

origin.

The NCBS facility requests the latest three quarterly sentinel health reports (third party

certification is a must) of the animal room from where the animals meant for shipping are

originally being maintained in the sender’s vivarium facility.

If the Health reports are compatible with import of animals in our high barrier facility: the

animals will be housed in quarantine as detailed below.

If the Health reports do not meet our SPF standards or if Health monitoring form the lab of origin

is not possible- Then the animals will need to be re-derived via surgical embryo-transfer. This

will need to be coordinated with the MGEF facility.

Quarantine procedures (= one month quarantine period after arrival of animals - even if

imported from JAX – without animal breeding possibilities)

Animals are taken in charge by the Animal Facility staff immediately upon arrival

(international shipments arrive in night time) and the transport boxes are taken inside the

facility through its service corridor to the quarantine room and are sprayed with 70% ethanol

before opening the box.

All the paper works attached to the box are collected; details on the labels are copied on to

the cage card.



Transport boxes are taken in and opened inside changing station of quarantine room and

animals are transferred to clean sterile cages using forceps and little feed from the box is

transferred to the fresh cage.

Animal are transferred to Individually Ventilated Cages (IVC) with fresh water and feed and

the empty transport boxes with bedding material are sent back to washing area.

Concerned PI and the Purchase department will be intimated on the animal condition.

Animals will be monitored in the quarantine room daily for any abnormalities or mortality

and are reported to the concerned PI.

After 27 days in quarantine, the institute veterinarian will submit the health status report of

imported animals to the regional quarantine officer and based on that report final quarantine

clearance certificate will be issued by the regional quarantine officer (RQO).

If there is any animal mortality, it will to be reported to RQO and a post-mortem examination

has to be done immediately and its report submitted to RQO for further action.

After 27 days, samples from sentinel animals (on contact with dirty bedding) from the

quarantine room is sent out for detail health report, based on the health report quarantine

animals are released for use.

Please note that no animal breeding is permitted during the quarantine period.

Estimated delays and timeline

- IAUC meets twice a month and can review proposals and proposal amendments as they

are submitted to its members

Note: FormB and <animal cage space request form> should be discussed with and approved by

the IAUC before complete project proposal submission to the IAEC.

- IAEC currently meets twice a year for final proposal review and approval (December and

June sessions)

- Estimated time for obtaining import license 10-12 weeks.

- Estimated time for obtaining health report after quarantine period is 10-12 days.



VI. AF Internal User Fees

VII. Publication Requirements

A. Acknowledging the NCBS Animal Facility in your publications and

conference presentations.

The facility is mostly financed through grants – these are dependent on publication records /

justifying the use of our facility to the funding agencies.

Publications not mentioning the NCBS High Barrier Animal Facility (NaMoR project) in their

acknowledgments therefore cannot be justified as publications having used the AF services and

that can directly impact the financial situation of the facility.

Please do acknowledge NCBS Animal Facility in your publications and oral presentations.

Our Recommended publication statement is:



< Animal work in the NCBS/inStem Animal Care and Resource Center was partially supported

by the National Mouse Research Resource (NaMoR) grant

(BT/PR5981/MED/31/181/2012;2013-2016) from the DBT >

We can provide you with HD versions of our NCBS AF logo as well. (See last page of this

document).

International authors Guidelines to publishing Animal Based research data and articles: Please do

read the one page ARRIVE guidelines (https://www.nc3rs.org.uk/arrive-guidelines) before

submitting your papers for publications. The ARIIVE guidelines have now been endorsed by

most of the major scientific journals, funding bodies and learned societies and describe all the

different items to include in your scientific research paper when publishing work involving the

use of laboratory animals.

B. Including an Animal care usage motion in your material & methods

Your publications should also clearly state in their materials and methods that:

< All the procedures that required the use of animals complied with Indian legislation concerning

vivisection and the use of genetically modified organisms, and the CPCSEA and IAEC Ethics

Committees on Animal Experimentation approved the protocols.>

Some Journals will also ask for the exact IAEC approved project ID# here.

C. Using Proper International animal strain Nomenclatures

Your material and methods should also refer to the mouse strains used for the study according to

the nomenclature defined by the International Committee on Standardized Genetic Nomenclature

for Mice.

Increasing number of journals are requesting that authors refer to their strains according to full

International Scientific nomenclature standards at least once in their material and methods. This

https://www.nc3rs.org.uk/arrive-guidelines



allows the international community to better understand which strains a given lab has used to

obtain their published results.

Please note that ACRC has imported many mouse stocks from Jackson Laboratories and we are

collaborating with JAX to obtain specific MTAs to allow Instem and NCBS labs to freely inter-

share some of these stocks from one institute to another- However this should be requested and

validated by the IAUC in advance in order for ACRC to make sure that the institute respects all

legal agreements and specific MTA clauses agreed with JAX for each particular stock.

Also note that original JAX or other imported stocks which have been bred at ACRC for more

than 5 generations can no longer be named and referenced as [/J] stocks bred from the JAX

facility- These stocks now need to referenced as being bred at NCBS and the ILAR approved

[Ncbs] animal facility code needs to me added to the stock full nomenclature in the publication

Material and Methods section. Please contact Lily for more clarifications regarding this.

There are still debates on how the new generation of Crispr targeted strains should be named but

a specific consortium is actually working on this and should deliver us a “Crispr nomenclature”

protocol in the coming months.

This mouse strain scientific nomenclature is also made mandatory in case any lab wants to

deposit their strains in any one of the international mouse repositories (JAX, EMMA, MGI,

RIKEN, and soon NCBS....).

The AF and IUAC can help you if you have trouble figuring out which correct nomenclature is

valid for the strains you are publishing with. JAX has also some useful information and tutorials

on this topic.

(https://www.jax.org/jax-mice-and-services/customer-support/technical-support/genetics-and-

nomenclature)

https://www.jax.org/jax-mice-and-services/customer-support/technical-support/genetics-and-nomenclature

https://www.jax.org/jax-mice-and-services/customer-support/technical-support/genetics-and-nomenclature



Contact & info: https://www.ncbs.res.in/research-facilities/acrc

Dr. Aurélie Jory (Lily): Head of the AF / Mouse Genome Engineering Facility:

Phone extension: 6563

Email: [email protected]

Dr. Mohan GH : AF - Head Veterinarian :

Phone extension: 6432

Email: [email protected]

Pr. Colin Jamora : IAUC chairman

Phone extension : 6759

Email : [email protected]

By the way: these awesome logos were designed by Sonal Kedia! Merci Sonal.





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