Richon ILAF Dec 2016 HO · 2017. 1. 2. · % change from pre-starved weight 7 5 7 12 22 Body...

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HEPs and HIPs:

Humane End Points Humane Interven3on

Points “Better Ethics with Less Pain and More Gain”

David Morton [email protected]

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HUMANE ENDPOINTS

Aim is not to cause avoidable suffering The earliest point at which an experiment can be stopped, or ought to be stopped

OR It can be regarded as the least inhumane endpoint.

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"Suffer ing"

AVOIDABLE

Practicewors t

NECESSARY

-----------------------------------------

(UNAVOIDABLE?)

bes t

(adverse effects) welfare

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HARMS/COSTS TO ANIMALS •  Direct as a result of the experiment •  Indirect as a result of the associated procedures e.g. housing, husbandry technical exper7se, etc

•  Must be minimum Necessary to achieve scien7fic objec7ve

•  NOT more i.e. Avoidable

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ENDPOINTS: SCIENTIFIC & HUMANE

Can refine on 2 fronts:

1. Animal suffering (Humane endpoints and Humane Interven7on points)

2. Experimental (Scien7fic endpoints)

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HUMANE ENDPOINTS

•  Aim of the experiment has been achieved

•  Animal not scien7fically useful due to physiological perturba7ons: i.e. data will be skewed by adverse effects

•  Animal not scien7fically useful due to psychological perturba7ons: i.e. data will be skewed by adverse effects

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HUMANE ENDPOINTS

•  Suffering higher than predicted – i.e. loss of propor7onality (H:B analysis)

•  Suffering too high -‐ ethically wrong •  Early (e.g. surrogate) endpoints should always be used whenever possible

•  Death rarely acceptable

ALTERNATIVE ENDPOINTS

•  When science can be achieved with less suffering e.g. Low temperature indicates imminent death

•  Biomarkers •  Analysis of other parameters to look for earlier endpoints which are s7ll scien7fically valid and objec7ves are s7ll obtained BUT Money / 7me needed to iden7fy what is going on

•  When it is secondary to main study objec7ves

(K.Ryder HO)

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Dr Jon Richmond

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OECD Guidance (2000, Monograph 19)

Humane Endpoint:

The earliest indicator in an animal experiment of severe pain, severe distress, suffering or impending death.

BUT now it is 2016

ALL MUCH TOO LATE??

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HOW DO WE KNOW WHEN ANIMALS ARE SUFFERING?

1.   How do we recognise suffering in all its forms e.g. distress, pain, discomfort, colic, nausea, ver7go, fear, anxiety, boredom, etc?

2.   When should we do it? •  Prospec7vely •  Retrospec7vely •  All the 7me

3.   Who should do it?

4.   Then use to develop HEPs HIPs, and allevia7on and avoidance strategies 12

NociceptionPain behaviors

SympatheticAdrenalMedulla

Hypothalamic-Pituitary

Adrenal cortex

Neurochemicalreceptors

dystress

FrightFlightFight

Mental distressFeelingsSufferingBoredomFrustra3on Happiness

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Starey coat, hunched, immobile, ears back, eyes closed, pinched abdomen,

huddling

Pain + Distress + Discomfort + Fear + ????

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Some considera7ons •  Is this animal normal? •  What physiological, pathological and psychological changes are happening? (3Ps)

•  Will it give reliable, repeatable and robust data? (Three Rs)

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LESSON LEARNED

MUST ALWAYS COMPARE WITH CORRECT CONTROL GROUP – NAÏVE ANIMALS

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HOW & WHY ASSESS ADVERSE EFFECTS?

Legal compliance Recogni7on of the abnormal Assessment of degree of abnormality OBJECTIVE assessment of impact e.g. for therapeu7c purposes, effects on science

Developing humane endpoints

ASSESSING WELLBEING

•  SIGN: observable by the doctor

•  SYMPTOM: reported by the pa7ent

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ASSESSING WELLBEING •  From a distance:

–  natural behaviour, – appearance

•  On handling –  bodyweight, – provoked behaviour, – clinical signs

Is it ‘normal’ for the individual, strain species?

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CLASSIFICATION OF SIGNS

Metric Signs -‐ a con7nuous variable quan7ty on a mathema7cal scale (linear or logarithmic) e.g. – Body weight – Body temperature – Heart rate – Respiratory – Behaviour

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CLASSIFICATION OF SIGNS Parametric signs can also be measured very accurately but oken on the basis of whether they are present or absent. They provide important informa7on just as valid and objec7ve as measurable signs e.g.

•  Appearance -‐ ruffled or starey coat, closed eyelids •  Response to given s7mulus -‐ withdrawal of a limb,

induced aggressiveness or vocaliza7on •  Posture: hunched, recumbent, boarded •  Clinical signs -‐ diarrhoea, dyspnoea, lameness

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SIGNS OF PAIN AND DISTRESS •  Changed: vocalisa7on, aggressiveness, posture, reac7on

to handling, movement

•  Absence of responses, prolonged recumbency, prolonged inability to reach food and water

•  Excessive weight loss, body condi7on (emacia7on)

•  Self-‐mu7la7on, open wounds or skin or corneal ulcera7on

•  Persistent diarrhoea, coughing, wheezing, dyspnoea

•  Polyuria, anuria, markedly discoloured urine

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SIGNS OF PAIN AND DISTRESS •  CNS signs: e.g. tremors, seizures, paralysis

•  Frank bleeding from orifices, dis7nct icterus, anaemia

•  Microbial infec7ons interfering with toxic or carcinogenic responses

•  Rapid weight loss, emacia7on, severe dehydra7on

•  Substan7al tumours

•  Treatment related effects indica7ve of impending death

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PAIN/WELLBEING ASSESSMENT IS SUBJECTIVE?

IS IT SUBJECTIVE BECAUSE OF: •  the clinical sign repeatability OR difficult to quan.fy?

OR IS IT SUBJECTIVE BECAUSE OF: •  the INTERPRETATION of the clinical sign

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GOOD ANIMAL HEALTH & WELFARE LEADS TO:

•  Good science : – more accurate data – more reliable data

•  Use of fewer animals •  Less animal suffering •  Greater economic value

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BAD ANIMAL HEALTH & WELFARE LEADS TO:

•  Poor science : – less accurate data – less reliable data

•  Use of more animals •  More animal suffering •  Poorer economic value •  Less science being carried out

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BENEFITS TO SCIENCE

•  Fewer losses with no data •  More accurate observa7ons with clinical relevance

•  Beper reproducible models

•  Repeated models vs novel work

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When you do not know e.g. new phylum, new species, new experiments, new context?

Absence of evidence is not necessarily evidence of absence

Cri3cal anthropomorphism/zoomorphism helps

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CLINICAL (OBSERVATION/ SCORE/ WELFARE ASSESSEMENT) SHEETS

Provides a way in which severity can be recorded for all to see, for observa7ons to be standardised and to set severity limits, humane endpoints and interven7on points.

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CONCLUSIONS Humane Endpoints and Humane Interven3on points are based on an analysis of recognisable clinical signs and other measurements

It is important to discriminate between avoidable and necessary suffering

HEPs and HIPs promote good science as well as good welfare

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CLINICAL OBSERVATION SHEETS

David Morton University of Birmingham

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CLINICAL (OBSERVATION/ SCORE/ WELFARE ASSESSEMENT) SHEETS

To be effec7ve oken have to be: •  Specific for each species •  Specific for each experimental protocol •  Signs are scored as present or absent •  They list common and KEY signs •  They can detail various ac7ons at the right 7mes

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By how much has it deviated from normality?

Is it possible to score/assess:

objec7vely (numbers -‐ metrics)

semi-‐objec7vely (parametrics)

subjec7vely (QBA/-‐Qual. Beh. Ass Free Choice Profiling)

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ASSESSMENT starts with a ‘new’ procedure to lab.

•  Consult the literature (???) •  *Contact other groups using the ‘model’ •  Predict signs -‐ use general sign and score sheet •  *Observe first group of animals individually •  Verify the literature (???) •  *Re-‐drak signs and if possible: define care, set scores, set humane endpoints (HEPs) and ac7ons (HIPs), ensure adequate training and competence of obersvers.

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ASSESSING ANIMAL WELLBEING •  From a distance:

–  natural behaviour, – Appearance

•  On handling: –  bodyweight – provoked behaviour, – clinical signs

Is it ‘normal’ for the individual, strain species? If not:

dbmorton, univ. birmingham, uk 38

Score sheet RAT No: 3 DATE STARVED: 9/5/99 ISSUE No: 8978STARVEDWEIGHT:

214 PRE-STARVED WEIGHT: 219

DATE 10-May 11-May 12-May 13-May 14-May 14-MayDAY 0 1 2 3 4 4

TIME (hr) 8:40 9:00 8:50 8:55 8:05 11:00FROM A DISTANCEFed mash/jelly/grain Y Y Y Y Y YInactive - - - + +Isolated - - - - -Walking on tiptoe - - - + +Hunched posture - - - + +Pinched face - - - + +Starey coat - - - + +/-Type of breathing N N N 120 L 70LNot grooming - - - + +Negative Red Light Response nd nd nd nd + +ON HANDLINGNot inquisitive & alert - - - + +Not eating - - - +/- +Not drinking - - - ? +Vocalisation on gentle palpation - - - - -Volume water drunk (average of rats in cage) ml 50 113 133 140 av 0Bodyweight (g) 204 209 203 192 170 168% change from pre-starved weight 7 5 7 12 22Body temperature ( oC) 37.5 37.4 37.6 32 35*Pale or sunken eyes - - - + +Dehydration - - - + +Distended abdomen/swollen - - - +/- +/-** Diarrhoea 0 to 3 (+m or +b) - - - - -Cage wet - +/- + + -Condition score 4 to 1* 4 4 3 2+ 2Saline given s/c - volume/sites? - - - 2ml/x2 -Blood sugar level nd nd nd nd ndNothing abnormal detected √ - - - -OTHER day 1 streptozotocin @65mg/kg animal killed

SIGNATURE: KL KL KL KLSpecial husbandry requirements:Feed irradiated diet and adapt animals to it 2-3 days prior to diabetes induction.Animals should be cleaned out twice daily.Two bottles of UV water should be provided for each cage and filled twice dailyDeprivation of water overnight may be sufficient to cause death bydehydration.Autoclaved diet must be provided.Scoring details* Breathing: R = rapid; S = shallow; L = laboured; N= Normal**0=normal ;1=loose faeces on floor; 2=pools of faeces on floor; 3 =running out on handling+m=mucus; b=bloodRefer to condition chart: 4: normal to 1: emaciatedHumane Endpoints and actions


RAT No: 3 DATE STARVED: 9/5/99 ISSUE No: 8978STARVEDWEIGHT:

214 PRE-STARVED WEIGHT: 219


TIME (hr) 8:40 9:00 8:50 8:55 8:05 11:00


FROM A DISTANCEFed mash/jelly/grainInactiveIsolatedWalking on tiptoeHunched posturePinched faceStarey coatType of breathingNot groomingNegative Red Light ResponseON HANDLINGNot inquisitive & alertNot eatingNot drinkingVocalisation on gentle palpationVolume water drunk (average of rats in cage) mlBodyweight (g)% change from pre-starved weightBody temperature ( oC)Pale or sunken eyesDehydrationDistended abdomen/swollen** Diarrhoea 0 to 3 (+m or +b)Cage wetCondition score 4 to 1*Saline given s/c - volume/sites?Blood sugar levelNothing abnormal detectedOTHERSIGNATURE:

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Special husbandry requirements:Feed irradiated diet and adapt animals to it 2-3 daysprior to diabetes induction.Animals should be cleaned out twice daily.Two bottles of UV water should be provided for each cageand filled twice dailyDeprivation of water overnight may be sufficient tocause death by dehydration.Autoclaved diet must be provided.

Scoring details* Breathing: R = rapid; S = shallow; L = laboured;N= Normal**0=normal ;1=loose faeces on floor; 2=pools of faeces on floor; 3=running out on handling+m=mucus; b=bloodRefer to condition chart: 4: normal to 1: emaciated

Humane Endpoints and actions1.Any animals showing signs of coma within the first24-48hrs will be killed.2.Any animals weighing less than the starting weight after 7 days willbe killed, or losing more than 20% than startweight will be killed3.Any animal showing tiptoe or slow ponderous gaitwill be killed.4.Inform veterinarian and PI if more than oneclinical sign occurs

Scientific MeasuresTissues to be kept - Kidney into formal saline


SCORING ROTE

‘ABNORMAL’ clinical signs are given a plus e.g. – dehydra7on, diarrhoea, not ea7ng

‘NORMAL’ clinical signs are given a minus – ea7ng, ac7ve

ONLY SCORE ANIMALS THAT ARE ‘NOT RIGHT’



TIME (hr) 8:40 9:00 8:50 8:55 8:05 11:00FROM A DISTANCEFed mash/jelly/grain Y Y Y Y Y YInactive - - - + +Isolated - - - - -Walking on tiptoe - - - + +Hunched posture - - - + +Pinched face - - - + +Starey coat - - - + +/-Type of breathing N N N 120 L 70LNot grooming - - - + +Negative Red Light Response nd nd nd nd + +ON HANDLINGNot inquisitive & alert - - - + +Not eating - - - +/- +Not drinking - - - ? +Vocalisation on gentle palpation - - - - -Volume water drunk (average of rats in cage) ml 50 113 133 140 av 0Bodyweight (g) 204 209 203 192 170 168% change from pre-starved weight 7 5 7 12 22Body temperature ( oC) 37.5 37.4 37.6 32 35*Pale or sunken eyes - - - + +Dehydration - - - + +Distended abdomen/swollen - - - +/- +/-** Diarrhoea 0 to 3 (+m or +b) - - - - -Cage wet - +/- + + -Condition score 4 to 1* 4 4 3 2+ 2Saline given s/c - volume/sites? - - - 2ml/x2 -Blood sugar level nd nd nd nd ndNothing abnormal detected √ - - - -OTHER day 1 streptozotocin @65mg/kg animal

killed

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THE ‘TEAM’ •  Animal care staff •  PIs, veterinary technicians, and all who carry out scien7fic procedures on animals

•  Project leaders and experimental design •  Sta7s7cians •  Veterinarians •  Ethics commipee members •  Enforcers •  Regulators

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Mouse rabies potency test

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Clinical signs in two mice Days after infectionMouse

NoClinical Signs

-1 0 1 2 3 4 5 6 7 8 9Sign-ruffled fur 0 0 0 0 0 + + + + + --slow movements 0 0 0 0 0 0 + + + + --circularmovements

0 0 0 0 0 0 0 0 0 0 -

-trembling 0 0 0 0 0 0 0 + + + --convulsion 0 0 0 0 0 0 0 + + + --lameness 0 0 0 0 0 0 0 0 + + --paralysis 0 0 0 0 0 0 0 0 0 + -

1

-agonal signs 0 0 0 0 0 0 0 0 0 + -Sign: --ruffled fur 0 0 0 0 0 0 + + + + --slow movements 0 0 0 0 0 0 + + + + --circularmovements

0 0 0 0 0 0 0 0 + 0 -

-trembling 0 0 0 0 0 0 0 + + + --convulsion 0 0 0 0 0 0 0 0 0 0 --lameness 0 0 0 0 0 0 0 0 0 0 --paralysis 0 0 0 0 0 0 0 0 0 + -

2

-agonal signs 0 0 0 0 0 0 0 0 0 + -

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Categories of clinical signs in rabies infec7on in the mouse

Clinical signs Score

Ruffled furHunched

1

Slow circling movements 2

Trembling shaky movementsConvulsions

3

LamenessParalysis

4

ProstrationRecumbency

5

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Scores of clinical signs in mice on a potency test of a batch of rabies

vaccine Day Mouse

1Mouse2

Mouse3

Mouse4

Mouse5

Mouse6

Mouse7

Mouse8

Mouse9

Mouse10

1 0 0 0 0 0 0 0 0 0 02 0 0 0 0 0 0 0 0 0 03 0 0 0 0 0 0 0 0 0 04 0 0 0 0 0 0 0 0 0 05 0 0 0 0 0 0 0 0 0 06 0 0 0 1 1 1 2 1 0 07 2 0 0 1 2 0 2 3 1 08 5 0 0 1 4 0 4 4 1 09 - 0 0 2 5 0 5 5 1 010 - 0 0 4 - 0 - 3 011 - 0 0 5 - 0 - - - 012 - 0 0 - - 0 - - - 013 - 0 0 - - 0 - - - 014 - 0 0 - - 0 - - - 015 - 0 0 - - 0 - - - 0

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FALSE POSITIVES -‐ too early an endpoint (N = 46)

BODY TEMP. FALSE(DEGREES C) (%)

35.5 19.5<35 4.3

<34.5 0<34.0 0

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BODY WEIGHT FALSE(%) (%)<20 45.6

<30 21.7<40 2.2<50 0

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CLINICAL SCORE FALSE(%)

1 47.82 23.93 2.17

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Days saved if a temperature of < 34.5oC is taken as the end point

Body Temp. oC Days saved

Cut off Mean Range

<34.5 2.17 1-7

STATISTICAL (UN)CERTAINTY AND HEPS, HIPS etc

•  Scien7sts accept a probability of 5% for their scien7fic endpoint measures, i.e. less than a 1 in 20 it could have occurred by chance.

•  They should therefore accept the same probability for HEPs and HIPs i.e. they need not be 100% accurate.

REMARKS

Can reduce animal suffering in research through: 1.  Earlier endpoints 2.  Surrogate endpoints 3.  Ensuring good science and preven7ng misleading

science 4.  Improved experimental design 5.  Beper communica7on and training

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•  Closer observa7on of animals by all staff as score sheets have indicated those 7mes that are cri7cal for the animal and for the experiment

ADVANTAGES OF ‘SCORE’ SHEETS

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•  Subjec7ve assessments of suffering by animal care staff and scien7sts are avoided.

•  Promotes more fruiwul dialogue, as evidence-‐based opinion becomes possible based on the clinical signs.

•  Helps experienced persons illustrate to less experienced persons why an animal is 'not right'.

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•  Consistency of scoring is increased as the guidance is clear and the scoring op7ons are limited.

•  Single signs or combina7on of signs can be used to indicate overall severity of the procedure.

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•  Helps to measure the impact of scien7fic procedures and aids comparing various alterna7ve models

•  Helps to indicate set points in a scien7fic procedure for interven7on (e.g. blood sampling, analgesia).

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•  The sheets help to determine the effec7veness of any therapy intended to relieve adverse effects

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ANIMAL CARE COORDINATOR

•  Checks that appropriate authori7es are in place and to marry up with what the scien7st actually intends to do that day to the animal(s);

•  to check the score sheet is appropriate before experiment begins;

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•  To know purpose of the experiment and the scien7fic objec7ves, and to become familiar with the scien7fic procedures to be carried out on the animals and the clinical signs that may occur;

•  To ensure all personnel (technicians, scien7sts) know how to use score sheets and can recognise the clinical signs and can interpret the signs clearly into humane end points;


•  To check that care staff not familiar with that experiment, say doing a weekend or holiday rota, are informed about those animals;

•  To liaise with PIs over the experiment eg. 7ming, numbers of animals, equipment, endpoints;

•  To update the score sheets based on new signs or combina7ons of signs observed



To report to the responsible persons (e.g. the PI, technician or the veterinarian) any concerns over the animals or personnel involved.

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WHEN CAN SUFFERING BE REDUCED?

Score sheets can be used to avoid, reduce or alleviate animal suffering in experimental procedures, especially those involving standardised protocols leading to less inhumane endpoints!

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EXAMPLES OF HEPS

1.  Procedure P 2.  Adverse effect AE 3.  Ac7on AC

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EXAMPLES OF HEPS

P: Repeated oral drug administra7on by gavage AE: may result in some drug solu7on entering the trachea and lungs and result in breathing difficulty manifested by laboured breathing and gasping for breath. AC: Animals showing signs of breathing difficulty will be killed.

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EXAMPLES OF HEPs

•  AE/AC: Any animal that shows a bodyweight loss of more than 20% shall be killed.

•  AE/AC: Any animal whose body temperature drops below 35C will be killed

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EXAMPLES OF HEPs P: Repeated needle injec7on can result AE: in 7ssue damage and infec7on. AC: Irrita7on caused by repeated injec7on will be avoided as much as possible by injec7ng small volumes and using different sites for repeat injec7ons.

AC: Rewarding rodents aker a procedure can reduce losses (M. Cuneen)

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EXAMPLES OF HEPs

•  P/AE/AC: Infec7on will be avoided by the use of asep7c techniques.

•  AC/AE: Animals will be inspected at least twice daily and any showing 7ssue damage or infec7on at sites of injec7on that deteriorates over a two day period will be killed.

SEVERITY LIMITS

HOW CAN WE SET LIMITS TO SUFFERING THAT ARE LOWER THAN SEVERE? MILD MODERATE SEVERE

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EXAMPLES OF HEPS

1.  Procedure P 2.  Adverse effect AE 3.  Ac7on AC

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EXAMPLES OF HEPs

•  AE/AC: Any animal that shows a bodyweight loss of more than 10/15/20% shall be killed.

•  AE/AC: Any animal with a tumour that is more than XXmm, or that has ulcerated or that show a body condi7on score of 2 or less will be killed

THE IMPORTANCE OF EXPERIMENTAL DESIGN

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EXPERIMENTAL DESIGN Non-‐ sta7s7cal

•  Tumour size -‐ test therapeu7c effect on small tumours before large ones

•  Use of minimal s7muli first e.g. analgesic tests •  Use of escapable s7muli when animal determines its own tolerance e.g. analgesic tests

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EXPERIMENTAL DESIGN Non-‐ sta7s7cal

•  Hierarchical approach e.g. Draize eye irritancy test •  Avoiding death as an endpoint •  What controls are needed

–  Studies on infec7on –  Historical data

•  Pilot studies –  ‘New’ husbandry –  ‘New’ method of anaesthesia, analgesia, euthanasia –  ‘New’ substance to be dosed – Will a change be bad for my research?

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EXPERIMENTAL DESIGN (Sta7s7cs)

•  Use of more powerful sta7s7cal methods e.g. blocking

•  Use of pilot studies to determine variance •  Interim review •  Use of pilot study data in main study •  Use of background historical data •  Care over controls exactly why might they be required e.g. pancreatectomy in diabetes research

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REFINEMENT Maximising benefits (research) e.g.

Sharing skills collabora7ve working

Sharing data

Sharing 7ssues

Ensure good prac7ces (educa7on, training,

competence)

Full repor7ng in scien7fic publica7ons/website

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NEW TECHNIQUES

•  Radiotelemetry: NB cost and harm of instrumenta7on •  Microarrays -‐ genomics, phylogenomics, proteomics, metabonomics

•  Magne7c Resonance Imaging (MRI) •  Positron Emission Tomography (PET) •  The Brain •  Biomarkers (e.g. XENOGEN) •  Virtual human

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Simultaneous Monitoring of Bacterial and GFAP Signals Following An7bio7c Therapy

Unt

reat

ed

GFAP

Lux

GFAP

Trea

ted

at 1

7h

Lux

– 106

- 4

- 4

- 2

- 8 - 6

- 8 - 6

- 2

– 104

– 105

- 4

- 4

- 2

- 8 - 6

- 8 - 6

- 2

– 104

– 106

- 4

- 4

- 2

- 8 - 6

- 8 - 6

- 2

– 105

– 106

- 4

- 4

- 2

- 8 - 6

- 8 - 6

- 2

– 104

3h 12h 18h 28h 41h 47h 65h

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CONCLUSIONS Clinical observa3on / score sheets help iden3fy types and 3mes of poor welfare and poor science

It is important to train all staff involved to recognise signs of poor welfare

HEPs, HIPs and severity limits can be developed using score sheets

Experimental design can help reduce animal suffering and promote good science

SEVERITY BANDING

WHICH ANIMALS SUFFER MOST

WHICH ANIMALS ‘SUFFER’ MORE?

1.  A social animal kept singly in a small barren cage for all of its life.

2.  An animal that has been subjected to an experimental procedure that causes severe suffering for 3 days.

3.  An animal that has been subjected to an experimental procedure that causes moderate suffering for 30 days.

4.  An animal that is deprived of normal amount of diet for 1 year and is maintained at 80% of its natural bodyweight.

WHICH ANIMALS ‘SUFFER’ MORE? 1.  A social animal kept singly in a small barren

cage for all of its life. 2.  An animal that has been subjected to an

experimental procedure that causes severe suffering for 3 minutes.

3.  An animal that has been subjected to an experimental procedure that causes moderate suffering for 3 hours.

4.  An animal that is deprived of normal amount of diet for 1 week and is maintained at 80% of its natural bodyweight.

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MILD, MODERATE, SUBSTANTIAL, SEVERE, INTOLERABLE?

An animal that has been subjected to an experimental procedure that causes considerable suffering for 10:

Seconds (Mild?) Minutes (Moderate?)Hours (Severe?) Days? Weeks? Months? This is not necessarily a ques7on about minimal suffering

to achieve the scien7fic objec7ve, it applies to clinical prac7ce!

HOW /WHEN WOULD YOU CATEGORISE SEVERAL MILD PROCEDURES AS

MODERATE?

Each technique could be only mild at any one 7me, but when done frequently may be more severe.

Suffering = (I1xT1) + (I2+T2) i.e. area under the curve? This may depend on the 7me between techniques 1-‐

n. Movement between project licences from a Mild

protocol to another Mild protocol may end up as Moderate to an animal?

Life7me experience to be assessed or just on the

experiment, or just that technique?

Richon T3

David Morton [email protected]

OTHER IMPORTANT AREAS AND COMMENTS

•  Severity Classifica7on, numbers, examples •  Assessment •  Gene7cally Altered Animals

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Severity

TWO COMPONENTS:

INTENSITY &

DURATION

Numbers of animals subject to severe severity in the EU (% -‐ Number)

•  AUSTRIA 10.3% 21,502 BELGIUM 14.8% 98,062 BULGARIA 26.5% 3,365 CYPRUS 0 DENMARK 1.5% ~3,000 ESTONIA 1.9% 115 FINLAND 4.9% 7,171 FRANCE 9.1% 161,829 GERMANY 6% ~168,000

•  GREECE 6.8% 2,881 HUNGARY 6.9% 13,862 IRELAND 40% 90,072 LATVIA 18.3% 2,510 MALTA 0

HOLLAND 2.7% 15,226 ROMANIA 0.05% 10 SLOVAKIA 1% 175 SPAIN 7.7% 62,579 UK 5% 185,481

Focus on models that may cause severe suffering

•  Stroke – cerebrovascular ischaemia •  Mul7ple sclerosis – Experimental Allergic Encephali7s •  Spinal Cord Injury – Paralysis •  Sep7c shock •  Myocardial ischaemia •  Primate neuroscience K.Ryder (HO)

Focus on models that may cause severe suffering

–  Severe cancer models, e.g. bone metastasis –  Irradia7on with recons7tu7on of bone marrow –  Infec7ous disease models –  Tamoxifen as inducer of gene func7on –  Vaccine tes7ng –  Some toxicology protocols –  Cerebral malaria in rodents –  Mul7-‐organ failure models –  Pancrea77s models

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Prospec7ve severity classifica7on Recital 22 (2010/63/EU)

“To enhance transparency, facilitate the project authorisa7on, and provide tools for monitoring compliance, a severity classifica7on of procedures should be introduced on the basis of es7mated levels of pain, suffering, distress and las7ng harm that is inflicted on the animals..”

Tools – retrospec7ve actual severity Recital 24 (2010/63/EU)

•  “When developing a common format for repor7ng purposes, the actual severity of the pain, suffering, distress or las7ng harm experienced by the animal should be taken into account rather than the predicted severity at the 7me of the project evalua7on."

Case-‐by-‐case assessment •  Type of species and genotype •  Maturity, age and gender of the animal •  Training experience of the animal with respect to the

procedure •  The actual severity of the previous procedures, •  Refinement of the procedure and housing, husbandry •  Care condi7ons •  Applica7on of humane end-‐points

S. Louhimes (EC)

Case-‐by-‐case assessment •  Type of manipula7on, handling, •  Nature of suffering caused by the procedure, incl. its

intensity, dura7on, frequency and mul7plicity of techniques employed

•  Cumula7ve suffering within a procedure •  Preven7on from expressing natural behaviour including

restric7ons on the housing, husbandry and care standards. S. Louhimes (EC)

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Case-‐by-‐case assessment Workman guidelines (2010): Single tumour, the mean diameter should not normally exceed 1.2 cm in mice or 2.5 cm in rats, or 1.5 and 2.8 cm, respec7vely, for therapeu7c studies. BUT makes no allowance for varia7on in the degree of pain or distress due to its loca7on e.g. axilla affects locomo7on is far more distressing than one located on back, or general physical and mental health, e.g.

a)  Weight loss (generally 15%)

b)  A fixed score on our score sheets

Severe suffering (K.Ryder HO)

•  Must be jus7fied •  Higher harms require higher benefits •  Frequent monitoring described in authorisa7on •  Clear welfare end points – How long suffering can con7nue – Not long las7ng severe suffering

Severe Classifica7on in Industry -‐ 1 (Regulatory Tes7ng)

–  For poten7al new medicines toxicity tes7ng (maximum tolerated dose studies)

– Vaccine efficacy and safety tes7ng – Acute toxicity studies (no longer required for poten7al medicines but may be required to support manufacturing e.g. transporta7on of chemical intermediates)

Severe Classifica7on in Industry 2 (Research)

•  Certain disease models may be classified as severe (e.g. some models in the following areas -‐ cardiac failure, chronic pain, oncology, infec7on, neuro-‐degenera7on, arthri7s) because they represent severe diseases in humans and that mimic as closely as possible these diseases for both pathophysiological and symptoma7c aspects.

•  These models are used to iden7fy poten7al new agents that are effec7ve against iden7fied targets linked to disease progression

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Industry View Sally Robinson (efpia)

•  Cause the least pain, suffering, distress or las7ng harm;

•  Death as the end-‐point to be avoided as far as possible and replaced by early and humane end-‐points.

•  Where death as the end-‐ point is unavoidable, the procedure shall be designed so as to: (a) result in the deaths of as few animals as possible; and (b) reduce the dura7on and intensity of suffering to the animal to the minimum possible and, as far as possible, ensure a painless death

Case Study: Refining Short Term Toxicity Studies

•  Short term toxicity studies are used to iden7fy early toxicity and set doses for longer term regulatory studies (and can provide alterna7ve informa7on to acute toxicity studies for medicines)

•  Usually severe classifica7on. Death is not an endpoint but provide informa7on on major adverse effects that are observed at the maximum tolerated dose (MTD)

•  MTD -‐ determined by clinical signs, reduc7ons in body weight. Limited published criteria/guidance on the intensity and dura7on of clinical signs that would op7mise the selec7on of an MTD

Case Study: Refining Short Term Toxicity Studies

•  Cross Industry/NC3Rs data sharing exercise on body weight loss in short term studies to assess the impact on the animal and study outcome* Data from 151 studies across species

•  Data supported limits for body weight loss of 10% in 7 days for rodents

•  Developed a alert/warning system for body weight loss in short term studies

•  Significant opportunity to reduce severity Sally Robinson (efpia)

Opportuni7es for refinement

•  Valida7on of humane endpoints for some forms of regulatory tes7ng

•  New technologies like bioimaging allowing a closer follow-‐up and earlier endpoints (smaller tumour size in oncology) Biomarkers

•  New strategy for tes7ng for convulsions: in vitro tes7ng on cells (ex. Ca flow through neurons),

•  zebra fish larvae, EEG allowing detec7on of signs before convulsions

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Opportuni7es for refinement

Obstacles •  Scien7fic constraints e.g. no validated alterna7ve

available in many cases

•  Acceptance by the competent authori7es for some regulatory studies (e.g. Pharmacopeia)

•  Technological constraints: some sensi7ve tools are not compa7ble with high numbers of animals

Effec3ve severity assessment (D Anderson)

•  Team approach -‐ with input from people with different

exper.se, experience and priori.es •  Good planning •  Educa.on and training of all personnel involved •  Appropriate, tailored recording systems •  Appropriate protocols for assessing behaviour and clinical

signs

Severity in rela3on to GA animals

•  Breeding of GA animals is defined as a procedure that needs to be authorized if the progeny is expected to experience pain, suffering, distress or las7ng harm (2010/63/EU Ar7cle 3 in conjunc7on with Ar7cle 17)

Severity in rela3on to GA animals Dr Be�ne Bert (Berlin Animal Welfare Officers Wkg

Gp May 2016)

•  To consider effects of gene7c manipula7on •  To consider effects of breeding and maintaining GA

lines.

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Principles for the assessment •  No addi7onally bred animals (surplus) •  No addi7onal distress for the animals but also the maximum

of informa7on •  Appropriate number of animals •  Altera7ons in rela7on to the wildtype •  Appropriate 7me points

CARE (Dr Be�na BERT) •  Nude animals with las7ng

hypothermia •  Immunodeficient animals

•  Hyperglycemia/ Hypoglycemia •  Disturbed maternal care

Lacta7on •  Glomerular nephri7s

•  High ambient temperature (>28°C), nes7ng material

•  Specific hygiene regimen •  Sufficient water intake and

cleaning of bedding material/ Control of blood glucose, glucose administra7on in drinking water

•  Foster mothers •  Urine monitoring

Other comments

•  Central data base for GA animals and their degree of severity •  Correct naming of GA animals •  Objec7ve criteria for the degree of severity •  Mild behavioural deficits = no adverse phenotype ? •  High standard housing condi7on = state-‐of-‐the-‐art or

refinement ?

7-‐Dec-‐16 112

Transgenic Mice •  Soon will be the commonest experimental animal? •  Already the commonest species •  Problems vary with genotype, but how do we improve their

exchange with welfare uppermost? •  Passports?

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7-‐Dec-‐16 113

Mouse Passports

•  Ongoing recording of informa7on from birth to death in different laboratories

•  Helps determine how to maintain and minimise any harms

•  Helps others to look aker them properly and avoids others making same mistakes

7-‐Dec-‐16 114

PASSPORTS

•  What to look for -‐ phenotype •  When and how to look for it •  Incidence •  Pathophysiology •  What to do about it •  Reproduc7ve and other data rela7ng to breeding and husbandry

‘Aggrega7on of marginal gains’ •  SEVERAL SMALL REFINEMENTS CAN HAVE A SIGNIFICANT

IMPACT •  Conduct a systema7c breakdown of the animal’s life

experience into component steps •  Iden7fy poten7al causes of suffering •  Research how these might be avoided or refined

Penny Hawkins (rspca)

‘Aggrega7on of marginal gains’

•  Issues with iden7fying severe suffering –  – Different interpreta7ons of severity –  – Reluctance to acknowledge severe suffering

•  Some reluctance to change protocols •  – Fear of introducing new variables •  Genuine uncertain7es e.g. use analgesics – but which

agents, and what protocols, and measures of success?

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Responsibility (H. Ehall) •  Change in A�tude

+ Study data versus sen7ent animals + Openness: the responsibility towards the public + “The dialogue with the other side” + Recruitment principles–who and why

•  + A Culture of Care + needs to start at the very top of the organiza7on + Open door policy–whistle-‐blowing policy/welfare cards + If in doubt -‐ anthropomorphism

•  + Animal welfare comes first + Collabora7on with compe7tors + Say it, do it, live it!

The Three Cs

1. Culture

2. Communica7on

3. Care

Martje Fentener van Vissingen (Roperdam)

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CONCLUSIONS

We should first focus on severe animal suffering The suffering of GA animals is an immediate concern, passports can help reduce it

Animal housing and husbandry are other areas in need of refinement and enhancement

Responsibility and a culture of care affect the aatude of staff towards animals and should be a priority

FOCUS ON SEVERE SUFFERING

•  Contact Dr Barney Reed •  [email protected]

•  For a COPY OF THE PRESENATIONS

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