Refinement of Experimental Procedures

Transcription

1 EU COST ACTION B24 LABORATORY ANIMAL SCIENCE AND WELFARE STOCKHOLM NOVEMBER 2004 Refinement of Experimental Procedures Jann Hau Chair of Comparative Medicine Uppsala University & University of Copenhagen

2 Refinement Good will Methods for objective assessment of suffering (pain, fear and stress) Studies of routine procedures and refinement initiatives Restraint, sampling, anaesthesia, surgery catherisation, metabolic cage housing

3 Important Tasks Ahead 1. Objective non-invasive assessment of (negative) impact of procedures 2. Objective assessment of the (positive) influence of gentling/conditioning and positive reinforcement training

4 Better Tools Imaging Telemetry Non invasive sampling of biomarkers Faeces Urine Saliva Sonophoresis

5 Better Tools Imaging Telemetry Non invasive sampling of biomarkers Faeces Urine Saliva Sonophoresis

6 Faeces PRO 1. Non-invasive 2. Integrated measure the area under the curve 3. Group Housing Possible CON 1. Time consuming extraction procedures 2. ALL faeces must be collected

7 Urine PRO 1. Good integrated measure 2. Simple Extraction procedures CON 1. Requires Metabolism Cages 2. Requires Single Housing

8 SALIVA PRO 1. Non invasive 2. Good human results 3. Easy to condition animals to salivate 4. In dog good correlation between serum cort and saliva cort CON 1. Small volume in small animals 2. Different qualities of saliva 3. Secretion rate may influence concentration

9 Stress assessment of routine procedures Chronic catherization Metabolic cage housing

10 Automated Sampling of Blood and Faeces to Study Distribution and Dynamics of Corticosteroids in the Rat Royo, Bjork, Carlsson, Mayo & Hau. Journal of Endocrinology 2004

11 Cort after Op 700 [corticosterone] hours Faeces (ng/ h kg bw) Serum ng/ ml) Royo et al 2004

12 Cort after Op 700 [corticosterone] hours Faeces (ng/ h kg bw) Serum ng/ ml) Royo et al 2004

13 IgA after Op ug IgA/ h kg bw hours Royo et al 2004

14 Correlation between Cort and IgA ug IgA / h kg bw before surgery ng corticosterone / h kg bw before surgery Royo et al 2004

15 Assessment of stress induced by common laboratory techniques Eriksson, Royo, Carlsson & Hau Experimental Physiology

16 Cort 3 days before, 3 days in m-cage, 3 days after ng corticosterone/ day kg bw days Daily Eriksson et al 2004

17 IgA 3 days before, 3 days in m-cage, 3 days after 60 mg IgA / day kg bw days Daily Eriksson et al 2004

18 BW 3 days before, 3 days in m-cage, 3 days after body weight (g) y = 3.75x y = 2.64x y = 5.25x ,5 1 1,5 2 2,5 3 3,5 days preassay assay postassay Eriksson et al 2004

19 Metabolic cages mildly stressful according to Conclusions Fecal corticosterone slight increase & IgA slight decrease The weight gain is reduced Metabolic in the metabolic cages cages

20 Pigs in metabolism cages Royo, Lyberg, Carlsson & Hau 2005

21 175 Fecal Cortisol Excretion 150 ng cortisol / d kg bw P1 M1 P2 M2 P3 M3 P4 M4 pig 1 pig 2 pig 3 pig 4

22 1400 Fecal IgA excretion 1200 ug IgA/ d kg bw P1 M1 P2 M2 P3 M3 P4 M4 pig 1 pig 2 pig 3 pig 4

23 Are pigs stressed by metabolic cages? Probably not very much

24 Conditioning Positive Reinforcement Training Reduces Stress of Annual Physical Examination for Chimpanzees at UTMDACC Lambeth, Hau, Perlman, Martino, Bernacky, Schapiro, 2004

25 Retrospective Study To examine whether positive reinforcement training reduced the stress associated with administration of anesthesia

26 Annual Physical Examinations Chemistry Hematology Blood Pressure measurements Method of Restraint

27 Methods of Administering Anesthesia (PR) Voluntary Present (PG) Present with dart gun (WS) Wire stick (DT) Dart (EC) Enter Cage

28 Chimpanzees males and 75 females 1 to 41 years old

29 Database (N=133) 618 records 5 physical exams for each animal ( ) Excluded data from animals which were pregnant, wounded, ill, chronically-infected or on a research protocol

30 Stress-sensitive Variables Total White Blood Cells (WBC) Segmented Neutrophils Blood Glucose

31 Overall Comparison of Voluntary vs. Involuntary Presents (N=133) Variables WBC Neutrophils Glucose Stressresp. p-value.002*.001*.000* Voluntary Injection (228 cases) NO MEANS Involuntar y Injection (390 cases) YES

32 Individual Health Record File History of Physical Health History of Exptl. Procedures History of Mental Health housing history social partners dominance rank in the pair/group compatibility with other animals affiliative behaviour response to training and human contact Hau & Schapiro: In The welfare of Laboratory Animals, ed. E Kaliste, Kluwer Academic Publishers 2004.

33 THANK YOU Collaborators: Senior staff in my Uppsala Lab: Dr Hans-Erik Carlsson and Dr Felix Royo SJ Schapiro (Austin, Texas) MA Suleman & N Moinde (Nairobi, Kenya) R Palme (Vienna, Austria) M Spångberg & K Westlund (SMI, KI Stockholm) R Murison, (Bergen, Norway) J Pamungkas & D Sajuthi, (Bogor, Indonesia)