Humane EndPoints (HEP)
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1 Humane EndPoints (HEP) aurora brønstad - university of bergen Definition of HEP OECD: A humane endpoint can be defined as the earliest indicator in an animal experiment of severe pain, distress, suffering or impending death When the animal suffer more than strictly necessary Organisation for Economic Co-operation and Development (OECD)
2 Types of HEP Severity cut-of (animal killed) Most common mice, rats, pigs, fish Reduce exposure Treat symptoms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
3 Ethics Causes of death in laboratory animals Circulatory collapse Organ failure Dehydration Starvation Is this a aim of our study?? Is this necessary for our study?? Legislation and ethics Animal welfare act General provisions on how animals should be treated ( 3) Animals have intrinsic value independent of the utilization value for people. Animals must be treated well and protected against unnecessary strain and stress. Definition of strain and stress applies to both actual strain and stress - as well as risk of pain:
4 Legislation and ethics Animal welfare act Use of animals in research... ( 13)... permission can not be given... if the animal are in risk of unnecessary suffering.... the stress to the animals should be reduced to minimum Plan Quality control Application form FOTS 0("+,2;$3"6'+(''! +1%'1$4-"%'$/%'()'-"24-./'2"'! ;2(4%82&-.',%/%-,&1'-"8'+('+1%'&(..%&3("' '()'2")(,4-3<%'$";2-/%8'-"8',%7,(8$&2;.%'8-+-=! :%6$.-3("/'-"8'%+12&/' 01-,-&+%,2/3&/'()'+1%'-"24-./'!"#$%"&%'()'%*+%,"-.')-&+(,/'' 5.-""2"6'%*7%,24%"+/'-"8'''',%/%-,&1'8%/26"' 9-"-62"6'+1%'1$4-"')-&+(,' Give end point criteria here!"#$%&'()*+",-./$%& )*-0(+-/($%&12%& (,3-4$& 5/",.67%&& 837$-.+.)7%&)(/(,-4$%& 9-.43(6-$,'7%& 6-4'.9-.+.)7& :/;-'./6(/,"+& 4./0-,-./$, </,(';(/,-./$,!"#$%&#$'()& $*')('7&(,4== >*"+-,7&4./,'.+& $7$,(6$%&?@A%& B!A%&$,",-$,-4$%& (C8('-6(/,"+& 0($-)/& Communication Coordination D.66*/-4",-./& D..8('",-./%& D..'0-/",-./&"/0& D./,'"4,$& Specify the actions you are going to take - Stop procedure - Treat symptoms - Kill the animal
5 Planning experiments Important questions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hat are you trying to achieve? When will you know you have achieved it? (or cannot achieve?) What effects are expected on the animals (Kathryn Ryder) Planning experiments Areas of special concern Monoclonal antibody production Cancer research* Acute toxicity testing Chronic toxicity testing Aging studies Pain research* Infectious disease studies*, vaccine trials * highly relevant at UoB
6 Planning experiments - ecample evaluate effect of antiparasitic drug Aim of study: approach Counting # wounds caused by parasites Alternative approach: Count number of parasites Stop the study when # statistic is significant Treatment of symptoms Parasites causes wounds, wound causes infections and infections delay growth. Delayed growth decrease economic outcome for the farmer (Kathryn Ryder) Intestine Fish ventricle Gas bladder liver columna muscle
7 Brain Tumor Noninvasive markers Biochemical substances in salvia, urine, faces Cortisol acute stress IgA chronic stress (Jann Hau)
8 Goal! No self dying animals - matter of honor Change in clinical profile Leptospira canicola Leptopspira ictero killed sick killed moribund found dead killed sick killed moribund found dead % % Nick Reed Objective killing-criteria - Score sheets Animal no: Date 0 (no) Messy fur Unnormal movement, Immobility CNS symptoms Open stitches Swelled abdomen abdomen Lordosis Weightloss > 10% X X X X More than 3 no 5 = Killing X X X 5 (severe)
9
10 Score sheets Must to be tailormade to the specific experiment include relevant clinical symptoms Larbour demanding Add it to your application Communicate it to your staff and train them how to use it Plot your results in an Excel sheet Individnummer Tumor Behandling Dødsårsak Alder ved død 1 tumor 1 beh A syk 90 2 tumor 2 beh B moribund 60 3 sham ingen død 90 4 control beh A moribund tumor 1 beh B syk 80 6 tumor 2 ingen syk 63 7 sham beh A syk 93 8 control beh B død tumor 1 ingen moribund tumor 2 beh A moribund sham beh B syk control ingen syk tumor 1 beh A død tumor 2 beh B moribund sham ingen syk control beh A syk tumor 1 beh B syk tumor 2 ingen død sham beh A moribund control beh B syk 135
11 Pivot table report (Excel) - Powerful tool Count of Dødsårsak Dødsårsak Tumor død moribund syk Grand Total control sham tumor tumor Grand Total tumor 2 suddenly moribund Average of Alder ved død Behandling Tumor beh A beh B ingen Grand Total control sham tumor tumor Grand Total tumor 2 dies earlier Can we use this information when we plan new experiments with these tumors??
12 Evaluation of animals clinical symptoms and behavior Experience in observing Animal behavior Increased activity Observe at night *Red light - increase activity Enrichment *APPENDIX A OF THE EUROPEAN CONVENTION (ETS NO. 123) Absence of evidence is not necessarily evidence of absence - Altman Commonly used but unreliable EP criteria in tumor research Animals are killed if they loose >20% BW, tumor increase > d=3 cm, or tumors are ulcerated. Weight loss Tumor size Ulceration
13 Weight loss Frequently suggested end point criteria Pro Easy to measure and quantify Objective/Unbiased Contra Less sensitive Weight gain caused by tumor growth or ascites (poor condition) Weight loss caused by tumor anti cancer therapy James Wallace End points related to fixed size should be carefully reviewed Tumor Size Tumor necrosis Tumor location and aggressiveness Relevant for tumors in restricted sites (cranium, eye, abdomen, muscle, footpad) Small tumor in mouth can interfere with food intake Can be a part of healing process, response to therapy Can cause anemia, dehydration, infection Untreated control animals Tumor on limb interfere with mobility James Wallace
14 Factors often under-estimated when dealing with HEP Training of personnel Animal care staff Recognize clinical symptoms Plan: When report, to whom report! Frequent observation of animals Frequent supervision 12. Care and supervision of animals...animals must be inspected daily,..., and as often as necessary... a shift system must be established... outside of normal working hours. 2 times a day? 4 times a day? every hour?? OWERALL GOAL : No unnecessary suffering Norw regulation of animal experimentation
15 Tools implementing HEP Clinical evaluation chart Decision making chart Training of personnel Frequent observation of animals Use of new technology to settle HEP Imaging MRI CT optical imaging Telemetry - wireless reporting technology Non-implanted, partiallyand fully implanted Blood pressure, heart rate, body temperature ++
16 Management Setting HEP: Team approach build a good research team! Principal investigator Research technical staff Graduate students Lab animal veterinarian Animal care staff Clinical pathologist Statistician? animal care committee members ethical committee animal welfare officers Better science by focus on HEP: Joseph Davison (chair CCAC council ) Data more reliable in less stressed animals Alternative methods may provides equally valid data Are there more relevant /important questions that can be addressed using EP? Studies with earlier EP are more time/cost effective Good science - animal welfare *Canadian Council on Animal Care
17 Planning experiments Difficult task: Predict HEP What is expected?? Knowledge of pathogen mechanism Pilot studies Clinical experience Same condition in humans Treatments used in humans Guidelines CCAC* Guidelines on choosing appropriate endpoint Preliminary pilot studies Determine required frequency of animal observation Define responsibility for animal observation When report Whom to report Training personnel in clinical animal observation
18 L e a r n i n g A I M Knowledge of why for research animal die if clinical symptoms are neglected Definition of HEP Important field of research for focus on HEP Legal requirements for implementing HEP in planning and performing animals experiments Knowledge of different types of HEP Use of imaging techniques, Telemetry, Biomarkers, score sheets Importance of observing conditions when animal conditions are evaluated (day/night active, use of red light, enrichment) Importance of training recognizing clinical symptoms and frequent observation Communication and organizing responsibility for the animals Limitations of weight loss, ulceration and tumor size as HEP Highlights from 2nd INTERNATIONAL CONFERENCE ON THE USE OF HUMANE ENDPOINTS IN ANIMAL EXPERIMENTS FOR BIOMEDICAL RESEARCH Berlin august 2005
19 Use of Imaging to setle humane endpoints
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