Back to Basics –

Accommodating the Non-Human Primate in Neuroscience Research

Andrew Winterborn, DVM

University Veterinarian/ Director, Animal Care Services

Queen’s University,

Kingston, ON

Overview

• NHP use in Neuroscience Research

• International Regulations

• Species-Typical Behaviour

• Primate Care and Use Program • Considerations for Housing NHPs

• Considerations for Personnel working with NHPs

• Considerations for Restraint and Training of NHPs

• Conclusion

NHPs in Neuroscience

• Neuroscience – Knowledge on how the brain works normally and under pathological conditions

• Close similarities b/w NHP and human brains

• EU – 1 in 3 citizens living with a brain disorder

• NHP only model to systematically study the relationship b/w activity of a single nerve cell and higher cognitive functions • 1981 Nobel prize for Medicine and Physiology – Visual Pathways

• 2007 CCAC animal use data reported 3,509 NHPs

• 803 NHPs used in PAU 1 and 2 • 58 Rhesus macaques

Animal Welfare

• Basic health and functioning - fed and housed, free from injury and disease, and relatively free from the adverse consequences of stress

• Affective states of animals - relatively free from negative states, including pain, fear, discomfort and distress, and capable of experiencing normal pleasures and comforts

• Ability to perform important types of natural behaviour - carry out normal patterns of behaviour, including normal affiliation with other animals

www.ccac.ca

Psychological Well-being

• Physical Health

• Ability to cope effectively with day-to-day changes in its social and physical environment

• Ability to engage in beneficial species-typical activities

• Absence of maladaptive or pathological behavior that results in self-injury or other undesirable consequences

• The presence of a balanced temperament and absence of chronic signs of distress

European Directive (86/609/EEC)

“any restriction on the extent to which an experimental animal can satisfy its physiological and ethological needs shall be limited to an absolute minimum” (European Union [EU] 1986, Article 5, paragraph b).

Species – Typical Behaviour Habitat - Rhesus

• Humans are the only primate that have a larger geographical range • Found ubiquitously throughout mainland Asia; from Afghanistan to India

and Thailand to southern China

• Observed – remote forests, agricultural regions, and urban areas

• Thrive in areas of deforestation – “weed” macaques

• 50% time foraging and moving between food sites

• Omnivores – Fruits and shoots, supp. with eggs, insects and small mammals

• Primarily terrestrial depending on region

Species – Typical Behaviour Social Structure - Rhesus

• Rhesus – relatively large troop size (avg. 20 – 30)

• Both sexes and all age classes

• Troops operate as closed matriarchal societies • Repel strangers with acts of aggression

• Females spend their lives in their natal troop

• Males leave the natal troop during adolescence

• Social interactions are driven in part by dominance (Gouzoules et al, 1998)

• Redirection of aggression

• Recruitment of agonistic aid – usually matrilineal kin

Species – Typical Behaviour Social Structure

• Increase in population density – increase in affiliative behaviour (grooming)

• Rely heavily on visual communication

• Facial signals ± vocalization • Threat - mouth in an “O” shape, direct stare, raising eyebrows quickly,

flapping the ears, a jerky head-bob etc

• Friendly – lip-smacking

• ~ 20% Rhesus are neophobic (Suomi ,1991)

• Activation of HPA axis

• Behavioural response – fear/withdrawal

Species – Typical Behaviour Macaca fascicularis

• “Ecologically diverse” – primary forests, secondary forests, and riverine and coastal forests

• Arboreous species – one study, never came to the ground except within 5m of a river

• Live in multi-male troops, ~ 30 members

• Males leave the natal group at sexual maturity • Strict linear dominance hierarchy among males – determined by age,

size and fighting skills

• Females, stable core of the group – close bonds with maternal kin

• Dominance hierarchy among females – grooming is common with lower-ranking females grooming higher ranking females.

Activity Budget of Wild Indian Rhesus Macaques in Different Habitats

Habitat Feeding Locomotion Resting Grooming Play Other

Temple 26.74 15.75 36.70 12.38 6.20 2.23

Urban 21.92 19.32 36.72 1.78 7.87 2.39

Pond 28.00 11.17 39.35 8.48 11.98 1.02

Roadside 31.26 14.28 34.66 9.80 9.55 0.45

Canal Side 25.00 19.84 40.03 5.93 7.10 2.10

Forest 33.60 24.14 35.08 2.15 3.49 1.54

Seth et al. 2001

Caveats to wild behaviour

1. Activity budgets of wild primates may be biased due to observational difficulties.

2. Generalising wild primate behaviour from geographical or temporal location to all members of the species is problematic.

3. Obtaining adequate samples in wild is rare.

4. Genetic differences between captive and wild populations.

Caveats to wild behaviour

5. Subjectivity of observations of captive and wild populations may question validity of comparisons.

6. Assumes that wild primates have adequate welfare.

7. Captive primates adapt to their environment, but this behavioural change is not necessarily indicative of decreased welfare.

(Adapted from Veasey et al (1996) and Shepherdson (1998))

Goals of Husbandry and Housing

• Increase the diversity and performance of normal (wild, desired) behaviours

• Increase behavioural choices

• Prevent abnormal behaviour developing (reduce

frequency of abnormal behaviour)

The presence of abnormal behaviour (e.g stereotypies) indicates that either currently, or at some stage in the past, the animal’s environmental conditions have been suboptimal (Mason, 1991)

Housing

• Expression of species-typical postures and locomotion

• Expression of species-typical behaviour • Promoted by cage design – reflect units of usable space

• Elevated platforms – upward flight

• Foraging opportunities

• Social Interactions • Perform species-appropriate activities – grooming, play, sleeping

huddles

• Individual housing • Practical and scientific reasons – maintain visual, auditory, or olfactory

contact with conspecifics

International Cage Size Comparisons

Weight Floor Area Min. Encl. Vol. Min. Vol. Animal Height

AWA 3 – 10 kg 0.4 m2 - - 76.2 cm

10 – 15 kg 0.56 m2 - - 81.28 cm

CCAC < 7 kg 0.4 m2 - - 90 cm

> 7 kg 0.75 m2 - - 120 cm

EU < 3 yrs * 2.0 m2 3.6 m3 1.0 m3 180 cm

>3yrs** 2.0 m2 3.6 m3 1.8 m3 180 cm

* An enclosure of minimum dimensions may hold up to three animals. ** An enclosure of minimum dimensions may hold up to two animals.

Macaques – Vertical Flee Response

• Long-tailed macaques in lower cage spend more time on perch than those in upper level cage.

• Long-tailed macaques show less stereotypy in vertically enhanced cages

• Rhesus macaques demonstrate the same responses

• All macaques – vertical flee response

• Hypothesized – vertical escape route to reduce stress

(Buchanan-Smith et al., 2004)

Housing

(Clarence et al. 2005)

Personnel

• Trained and observant personnel • Essential for maintaining colony

• Best position to note signs of illness, injury or distress

• Aware of safety precautions • Prevent physical injuries and disease transmission between themselves

and the animals

• Standard Operating Procedures

• Dedicated NHP staff • Unfamiliar caregivers can be stressful to NHPs

• Communication • Integrated program between researchers and animal caregivers

Daily Care

• Joint effort between caregivers and laboratory staff • Note deviations from physical and behavioral norms

• Caregivers – daily interaction with animals • Inspection of animals - hair condition, gait, appetite, activity level,

injury and any abnormal behaviour

• Health Score – Caregivers and lab staff

• Routine • NHPs are neophobic

• Communication

Restraint and Training

• Restraint for examination and treatment mostly unavoidable • Restraint – brief as possible and carefully tailored to the species

• Train for routine procedures • Move into holding pens, injections and oral administration

• Basic principles for training • Appropriate response needs to be apparent to the subject

• Training should proceed in steps with attainable goals established

• Training session should continue until goal is accomplished

• Procedure should be routine

• Routine can reduce overall stress

• Allow for latitude in performance

Restraint and Training

Conclusion

• Macaques form complex social structures

• Adapt to diverse habitats

• Promote normal behaviours and prevent abnormal behaviours

• Housing and care that better meets the psychological well-being/ethological needs of NHPs • Address species-typical behaviour

• Training for routine behaviours

• Trained personnel

• More research is required into the stress-reducing potential of husbandry/housing strategies

Scientific Committee on Health and Environmental Risks

The need for non-human primates in biomedical research, production and testing of products and devices

“Refinements in the housing and care of animals that meet their mental needs will result in better animal welfare and better models………Poor housing and care standards will cause avoidable suffering and are likely to produce animals less suitable for studies, particularly in the neurosciences.”

http://ec.europa.eu/environment/chemicals/lab_animals/pdf/scher_o_110.p

df

References

• Clarence, W., et al. (2006) Use of Enclosures with Functional Vertical Space by Captive Rhesus Monkeys (Macaca mulatta) Involved in Biomedical Research. JAALAS 45(5) 31 - 34

• ILAR, The Psychological Well-Being of Nonhuman Primates (1998). National Academic Press.

• NC3Rs Guidelines: Primate accommodation, care and use. (www.nc3rs.org.uk)

• Reinhardt, V., Refining the traditional housing and handling of laboratory Rhesus Macaques. (www.awionline.com)

References cont.

• SHERC, The need for non-human primates in biomedical research, production and testing of products and devices. (http://ec.europa.eu/environment/chemicals/lab_animals/pdf/scher_o_110.pdf

• Seth el al. Indian Rhesus Macaque: Habitat, Ecology and Activity Patterns of Naturally Occurring Populations www.wii.gov.in/envis/primates/downloads/page68indianrhesus.pdf

• Buchanan-Smith HM, et al., (2004) What factors should determine cage size for primates in the laboratory? Animal Welfare (13)S197-201

• Rennie A. & Buchanan-Smith H. (2006) Refinement of the use of non-human primates in scientific research. Part I: The influence of humans. Animal Welfare 15:203-213.

• Rennie A. & Buchanan-Smith H. (2006) Refinement of the use of non-human primates in scientific research. Part II: Housing, husbandry and acquisition. Animal Welfare 15:215-238.

• Rennie A. & Buchanan-Smith H. (2006) Refinement of the use of non-human primates in scientific research. Part III: Refinement of procedures. Animal Welfare 15:239-261.

• Gouzoules et al., (1998) Agonistic screams and the classification of dominance relationships: Are monkeys fuzzy logicians? Anim Behav (55)51-60

• Suomi, SJ, (1991) Up-tight and laid-back monkeys: Individual differences in the response to social changes. In: S. Brauth, W. Hall, and R. Dooling, eds. Plasticility of Development. Cambridge: MIT Press. P. 27-56

M A S T E R T I T L E

Questions ?