How to write an application for animal experiments22/03/2017

Ethical Committee for Animal Experiments

• In Dutch: Ethische Commissie Dierproeven - ECD

• Regulated by law Royal Decree (Koninklijk Besluit) 29/05/2013

• Assignment

– Evaluation of the planned and executed animal experiments: i.e. application & retrospective analysis

– Formulate criteria for the ethical use of laboratory animals

– Offer advice to the labs concerning ethical aspects of animal experiments and concerning supervisory authority

Ethical Committee for Animal Experiments

• Composition:

– Min. 7 members

– Currently: 18 members + 1 advisory member + 1 secretary

• Several different competences & expertise:

– Ethics

– Alternative methods to animal experiments

– Research & research methods

– Statistics

– Animal wellbeing

– Veterinary

• Not all experts/researchers => No technical terminology

General evaluation criteria

• Scientific value and relevance

– Is the project required on scientific or educational grounds or legally

imposed?

– but NOT scientific project application

• Do the objectives of the project justify the use of animals?

• Is the project designed as humanely as possible?

• 3 R’s

General evaluation criteria

• Relevance & justification

• Implementation of the principles of 3 R’s: Replace, Reduce, Refine

• Methods (experimental, observational, statistical) used to reduce the number of

animals and their pain, suffering and fear

• Preventing non-justified duplication of procedures

• Re-use of animals & the impact on the degree of severity

• Expected degree of severity

• Use of anesthesia, painkillers, other means of reducing the animal’s inconvenience

and suffering, from birth to death

• Care and housing of the animals

• Use of humane end points

• Methods for killing animals

• Proficiency of the persons involved in the project

Application procedure• 2 documents:

Application form

+ Non-Technical Summary (Niet-Technische Samenvatting – NTS)

• Download latest version from ECD webpage for each new application

• Submit signed paper version + electronic version to ECD secretariat

• 1 deadline each month, 4 weeks prior to ECD-meeting

– Next: 18 April

• Approval => Licence (“Vergunning”)

– only valid for approved experiments.

– changes to the project => should be reported & approved by ECD

Application Form

P. 1Preliminary Information

Project number

• e.g. 18-220-1

• to be used in all

communication, ordering

animals & cage labels

Part 1.General Project Information

Part 1. General Project Information• Title Project vs Title NTS

• Responsible Project Manager (“Proefleider”) responsible for design, execution and follow-up of experiments + Contact person for the ECD

• Sponsoring / Funding, relevant to assess the scientific value

• Duration:

– Max. 5 years

– Expected start date not prior to ECD meeting, earliest possible start is the date of the ECD approval

– End date: no procedure can take place after this date

• Severity:

– highest level of all procedures; severity per experiment in part 6

– No P-scores

– Annex 5 of Royal Decree for definitions and examples

Part 2.Users

• Coordinates of the responsible lab director with LA permit (“LA erkenning”)

• Partner lab:

if 1 or more procedure is done in another lab than the main LA lab, e.g. in GFAN

NOT if animals are only being kept elsewhere

Part 3.Staff

• VUB staff should be mentioned on the LA permit

• All persons involved (also external) need the appropriate certificates, Royal Decree Annexes 8-11

• For certificates obtained abroad, the Department of Animal Wellfare (“DienstDierenwelzijn”) can approve the equivalence of the training

• Follow-up: it’s possible these are the same persons. Contact details required

Part 4.Project

• General description

≠ overview of experiments, part 6

• In non-technical terms for the entire committee to understand

• Purpose:

Multiple choices are possible

Fundamental research + Translational/Applied research

Students observing?

=> Higher education or training

Part 5.Animals

• Exact number, no ranges

• Justification for the number of animals is to be specified for each experiment in part 6

• Breeding only possible with specific LA breeding permit (LA2230)

Part 5.Animals - Exemption

Exemption required for

animals not specifically

bred for use in experiments

Part 6.Experiments & Procedures

Diagram example

Title project

Experiment 1: Stimulation of tumor-specific immune responses in vivo

upon immunization with in vitro treated tumor cells.

Procedure 1: Immunization of C57BL/6 mice at

left flank

Procedure 2: Challenge with

viable tumor cells at the right flank,

5 days after immunization

Procedure 3: Tumor growth

will be followed on a daily basis

Experiment 2:…

Procedure 1:…

Part 6.2.b.Description of experiments

Description of experiments - Example

Title of experiment:

Stimulation of tumor-specific immune responses in vivo upon immunization with in vitro treated tumor cells.

Relevance & Justification:

Tumor cells will be treated in vitro. These will be used for immunization of C57BL/6 mice (left flank). When genuine ICD is induced upon treatment of the tumor cells, a tumor-specific immune response should be elicited. To evaluate this, mice will be challenged with viable tumor cells at the right flank, 5 days after immunization. Tumor growth will be followed on a daily basis. This experiment will be performed twice to ensure reproducibility (n = 2).

Description of experiments - Example

Number of experimental groups and animals per group:

The treatment groups are mice receiving an intradermal injection of 0.5 x 106 tumor cells (in 50 µl PBS) treated with:(1) classical RT (positive control)(2) β-particle TRNT using 177Lu-Nbs (3) α-particle TRNT using 211At-Nbs(4) non radioactive labeled Nbs (negative control 1)(5) β-particle TRNT using 177Lu-Nbs (negative control 2)(6) α-particle TRNT using 211At-Nbs (negative control 3)

Description of experiments - Example

Justification of the number of animals. => Power analysis!

Number of animals per test condition is 5, based on the following parameters: valueto compare against = tumor size of 1500 mm3 in mice treated as described incondition (4-6) (expected to be reached at day 20), expected tumor size in the testconditions (1-3) is 1000 mm3 with a standard deviation of 250 mm3 (the conditionused for comparison = equal variance), 95% confidence interval (with Bonferronicorrection for 6 groups), resulting in a power of 80%. Survival will be plotted in aKaplan-Meier curve and analysis will be performed using the pair wise Mantel-Cox(log rank) test. Thus for this experiment we require: 5 (mice per condition) * 6(treatments) * 2 (number of repeats) = 60 mice C57BL/6 mice.

Describe in detail all the procedures/actions; e.g. volume and frequency of sampling, etc.Procedure 1: Immunization of C57BL/6 mice at left flank (SOPxxx). Procedure 2: Challenge with viable tumor cells at the right flank, 5 days after immunization (SOPxxx). Procedure 3: Tumor growth will be followed on a daily basis (SOPxxx).

Part 6.2.c.Severity & Animal Wellbeing

• Severity: highest degree

• No long-term severe degree

• Approval of vet required for re-use & only possible in case of max. moderate severity

Part 6.2.c.Severity & Animal Wellbeing

Painkillers & sedatives:

• literature and/or vet

• dose, frequency

Part 6.2.d.End of experiment

Both options (euthanasia & keeping alive)are possible after the same experiment

Part 7.3 R’s: Replace

• Also in NTS

• Demonstrate that researcher is aware of and has considered alternative methods

Part 7.3 R’s: Reduce & Refine

• Justification of the number of animals => part 6

• Species: lowest possible.

• Avoiding duplication through collaboration & staying up-to-date on the latest developments

• Duplicate = exact copy of experiment with the exact same group/condition

• Own experiments or from other labs

Part 8.Safety

Signatures

Non-Technical Summary NTS

Why?

• Imposed by decree

• Information on the objectives of the project, including the predicted harm and benefits, animal species and number of animals

• Proof that the replacement, reduction and refinement have been considered.

• Intended to inform broad public

• Published on website of authorities

• Needs to be submitted and updated together with ECD application

Target audience

• To inform in correct and concise way why the research with animals is necessary

• Not to convince the opponents of animal research

NO technical terminology!

Imagine someone from your family or friends:

• Without pronounced opinion about animal experimentation

• Eats meat, has a pet

• Has no (bio-)medical background

• Has education level at which VRT news focuses

General rules• Languages accepted: Dutch, French, German

– NO English

– Possibly translation needed of the project title

• Use the template provided & submit in excel

• Respect the indicated maximum of characters

• Duration: max. 5 years

• Format dates: dd/mm/yyyy

• Severity: no P-scores, use EU categories (in Dutch: terminaal, licht, matig, ernstig)

• Anonymous/confidential: no reference to names/persons/labs, to safeguard IPR

Tips• Limit technical terminology, use explanation or analogy to avoid

jargon

e.g. use “under the skin” rather than “subcutaneous”

• Leave out information/words that the reader doesn’t understand

e.g. “In dit project wordt onder verdoving met ip injectie van nembutal bloed genomen via de retroorbitale sinus en wordt het dier nadien geëuthanaseerd door het leegbloeden via de aorta.”

• Avoid latin words, e.g. Macaca mulatta = rhesus monkey

• No strains (species & number of animals)

e.g. Mus musculus Balb/c/hCD20-TamCre Tg => mice

• Avoid abbreviations, acronyms

• Use short sentences (as you would orally explain to someone)

• State things as they are, do not gloss over

• Non-invasive techniques may be stressed

• When the animals are not euthanized at the end of experiment, mention this and their final destination

• Make sure to show that you did a cost-benefit analysis

• Do not state the obvious

e.g. “Proper care” of the animals is evident

Tips

ECD ContactECD-secretariaat@minf.beECD-voorzitter@minf.beRoom D.327www.vub.ac.be/onderzoek/beleid/proefdierencommissie