Table 2. Generalised welfare assessment protocol for laboratory-housed macaques.

Assessor: ___________ Animal ID: ___________ Date: ___________
Indicator	Score	Scoring criteria	References
Animal-based – score for each animal
1. Self-harm behaviour E.g. on inspection or recorded in daily logs. Where seen, more frequent and detailed follow-up observations can be made to assess incidence, severity, and impact.	0	Self-harm behaviour not observed; individual not known to self-harm	Reinhardt & Rossell, 2001; Novak 2003, 2021; Polanco et al., 2021
	1	Self-harm behaviour observed, without physical injury; individual known to self-harm occasionally (e.g. self-biting, self-hitting, eye poking, head-banging, hair plucking/pulling)
	2	Physical injuries present, consistent with self-harm behaviour (e.g. abrasions, lacerations, eye trauma)
2. NHP induced injuries Use 2a and 2b. 2a. Injuries E.g. on inspection or recorded in daily logs.	0	No injuries present	Beisner et al., 2019; Crast et al., 2021
	1	Minor injuries present, consistent with fighting; may or may not require veterinary intervention/treatment (e.g. abrasion/blunt trauma, puncture wound, skin laceration)
	2	Severe injuries present, consistent with fighting; veterinary intervention/treatment necessary (e.g. deep or multiple laceration/s, skin puncture + muscle involvement, inflamed or infected wound, bone exposure, degloving; signs of pain such as grimacing, hunched posture, guarding of limb)
2b. Lameness E.g. on inspection or recorded in daily logs.	0	No signs of lameness or imbalance when moving; unimpaired/normal locomotion	Lewis & Colgin, 2005; Wren et al., 2013
	1	Signs of lameness or imbalance when moving (e.g. limping or favouring limb, slow locomotion or uneven rhythm, unable to keep up with the group, guarding of limb); interfering with normal locomotion, may or may not require veterinary intervention/treatment
	2	Persistent or long-term signs of lameness or imbalance when moving; inability to locomote normally; requires veterinary intervention/treatment
3. Appetite Choose the most appropriate method for your context: 3a, 3b or 3c for eating, plus 3d or 3e for drinking. 3a. Hand feeding For animals that are typically comfortable taking food from the hand.	0	Takes preferred food	Keeling & Wolf, 1975; Wolfensohn & Honess, 2005; Smith et al., 2006; Prescott et al., 2010; Association of Primate Veterinarians, 2019
	1	Takes only small amounts of preferred food
	2	Refuses to take preferred food; reluctant to come forward
3b. Eating habits E.g. on inspection at feeding time; or recorded in daily logs.	0	Observed to eat normally; evidence of food consumption (e.g. crumbs/scraps)
	1	Observed to not eat; no evidence of food consumption (e.g. no crumbs/scraps or food intact)
	2	Repeatedly observed to not eat (i.e. for more than one day); protracted lack of evidence of food consumption (e.g. no crumbs/scraps or food intact)
3c. Food consumption E.g. in the last 24 hours; weigh food or count biscuits/residues); can be recorded at the group level.	0	Normal amount of food consumed
	1	Reduced food consumption (e.g. 25-75% relative to baseline; or 1 SD below the mean)
	2	No (or very little) food consumed; reduced faecal output
3d. Drinking habits E.g. on inspection at feeding time; or recorded in daily logs.	0	Observed to drink normally; evidence of water consumption (e.g. approach water source)
	1	Observed to not drink; no evidence of water consumption (e.g. does not approach water source)
	2	Repeatedly observed to not drink (i.e. for more than one day); protracted lack of evidence of water consumption (e.g. does not approach water source)
3e. Fluid consumption E.g. in the last 24 hours; weigh water bottle; can be recorded at the group level.	0	Normal amount of fluid consumed
	1	Reduced fluid consumption (e.g. 25-75% relative to baseline; or 1 SD below the mean)
	2	No (or very little) fluid consumed; dry faeces
4. Body weight Use 4a or 4b (age dependent), plus 4c. Note weight loss is expected for some research protocols. 4a. Body weight change (for adult animals) E.g. relative to previous day or week.	0	No change in body weight	Wolfensohn & Honess 2005; Smith et al., 2006
	1	<10% change in body weight (or 1SD from expected for individual of that age) (Weight loss over a certain period may not be a concern in overweight animals, so check body condition score, 4c)
	2	>10% change in body weight (or 2SD from expected for individual of that age)
4b. Growth rate (for young, growing animals) E.g. using colony-specific growth curves, or those in the published literature.	0	Appears to be growing normally (e.g. body weight within normal range for age and sex, growth is following the relative centile)	Van Wagenen & Catchpole, 1956; Prescott et al., 2010
	1	Deviation from expected growth rate (e.g. body weight outside normal range for age and sex, deviation from the individuals’ normal growth trajectory, crossing centiles)
	2	Ceasing to grow normally (e.g. body weight far outside normal range for age and sex)
4c. Body condition score Condition scoring may be performed non-invasively through observation, or during clinical examination by palpating the thoracic and lumbar vertebrae.	0	Body condition score of 3 (normal/optimal)	Wolfensohn & Honess, 2005; Clingerman & Summers, 2012
	1	Body condition score of 2 (underweight/thin) or 4 (overweight/heavy)
	2	Body condition score of 1 (severely underweight/emaciated) or 5 (obese/grossly obese)
Sub-total: _ /14
Resource-based – score for the cage
5. Social enrichment Use 5a and 5b. 5a. Social condition Regardless of whether an exemption from social housing is approved by the IACUC/AWERB. In very rare cases, individuals may thrive without a social partner.	0	Continuously socially housed with one or more compatible conspecifics in the same cage/enclosure	Schapiro et al., 1996; Lutz & Novak, 2005; Gilbert & Baker, 2011; Baker et al., 2012, 2014; DiVincenti & Wyatt, 2011; Hannibal et al., 2017; Cassidy et al., 2020
	1	Intermittent social housing (during part of the day/week)
	2	Single housed (no physical contact but visual and olfactory contact provided); OR protected contact (separation of individuals via a barrier that permits social contact but not entry into each other’s cage/enclosure)
	3	Social isolation (no sensory contact with conspecifics).
5b. Social behaviour E.g. on inspection or recorded in daily logs.	0	Frequent prosocial/affiliative interactions observed between social partners; pair/group appears stable
	1	Both affiliative and agonistic interactions observed between social partners; may be some signs of pair/group instability
	2	Frequent agonistic interactions and few affiliative interactions observed between social partners; pair/group appears unstable; OR animal singly housed
6. Caging environment Use 6a and 6b. 6a. Cage dimensions Assessed against, e.g. ILAR Guide; ETS 123; Directive 2010/63/EU.	0	Provided with more than the regulatory/accreditation-related minimum space allowance (e.g. via large indoor enclosure, outdoor enclosure, access to additional exercise enclosure/play pen)	Reinhardt et al., 1996; Buchanan-Smith et al., 2004; Griffis et al., 2013
	1	Provided with the regulatory/accreditation-related minimum space allowance
	2	Provided with less than the regulatory/accreditation-related minimum space allowance (e.g. metabolism cage)
6b. Vertical space	0	Housed in cage floor to ceiling high, with adequate high perching, verandas, etc. to allow all occupants to move to heights about human eye level	Reinhardt, 1992; Nakamichi & Asanuma, 1998; Reinhardt, 2003; Clarence et al., 2006; Maclean et al., 2009; Gottlieb et al., 2014; Lutz & Brown, 2018
	1	Housed in cage floor to ceiling high, with limited high perches, verandas, etc., meaning not all animals have access
	2	Housed in double-tiered (1-over-1) caging that lacks perching at or above human eye level
7. Physical enrichment (including cage furniture) Use 7a and 7b. 7a. Provision of physical enrichment	0	Complex environment, with ample physical enrichment provided including structural enhancements (e.g. swings, ladders, shelves, tyres, hammocks, perches) that allow for species-typical locomotion (climbing, leaping, running, etc.), visual barriers, and manipulanda (e.g. toys, mirrors, wood blocks); structural complexity allows for as much of the housing to be used as possible	Bryant et al., 1988; Turner & Grantham, 2002; Honess & Marin, 2006; Waitt et al., 2008, 2010; Griffis et al., 2013; Descovich et al., 2019
	1	Limited physical enrichment provided (e.g. perches with toys)
	2	No physical enrichment provided
7b. Use of physical enrichment E.g. on inspection or recorded in daily logs. Note use of physical enrichment can vary with age.	0	Observed to frequently interact with physical enrichment in species-typical, positive way
	1	Observed to occasionally interact with physical enrichment in species-typical, positive way
	2	No interaction (or abnormal interaction) with physical enrichment
8. Food enrichment	0	Food presentation encourages daily and extended bouts of species-typical foraging behaviour (e.g. scatter feeding of fine forage mix into floor substrate; feeding fresh browse and edible plants from approved sources; utilising puzzle feeders, which require considerable time, manipulation, and fine motor skills for retrieval of food)	Chamove et al., 1982; Boccia 1989a,b; Byrne & Suomi, 1991; Reinhardt, 1994; Doane et al., 2013
	1	Food presentation encourages species-typical foraging behaviour, but such opportunities are not daily, nor extended (e.g. feeding whole fresh produce)
	2	Food presentation does not allow species-typical foraging behaviour (e.g. cafeteria-style presentation in bowls)
Sub-total: _ /15
Staff-based – score for the individual or facility, as appropriate
9. Positive reinforcement training (PRT) Use 9a and 9b. 9a. Individual training performance E.g. on inspection or recorded in daily logs.	0	Individual is trained, using positive reinforcement, to voluntarily cooperate with the scientific, veterinary, and husbandry procedures it is exposed to; it reliably cooperates, showing a high degree of compliance and few signs of distress	Prescott et al., 2005; Prescott & Buchanan-Smith, 2007; Perlman et al., 2012; Baker, 2016; McMillan et al., 2017
	1	Individual is trained, using positive reinforcement, to voluntarily cooperate with the scientific, veterinary and husbandry procedures it is exposed to; however, it does not reliably cooperate and shows signs of distress
	2	Individual is not trained to voluntary cooperate with procedures
9b. PRT programme		Characteristics of a high-quality programme: a. A specialist in behaviour modification was involved in development of the training programme b. One person has overall responsibility for the training programme (training coordinator) to ensure a consistent approach c. Animals are trained by dedicated staff members, who possess a high degree of training knowledge and competence d. Training procedures are documented in SOPs/protocols e. Methods are predominantly based on PRT; where negative reinforcement is used, it is in combination with PRT and only where PRT alone has failed f. The training programme allows sufficient time for progressive habituation to conditions, techniques, and procedures before data collection begins g. Records of individual training performance are kept and regularly reviewed; training is tailored to individual differences in learning
	0	Programme includes more than four of the above characteristics
	1	Programme includes two to four of the above characteristics
	2	Programme includes less than two of the above characteristics
10. Behavioural management programme		Characteristics of a high-quality programme: a. Incorporates enrichment interventions that satisfy a range of needs in the social, locomotory, sensory, cognitive, and food-based domains b. Offers novelty (objects, space, activities) to stimulate and provide new challenges c. Provides animals with a degree of choice and control in their environment d. Incorporates regular monitoring for behavioural signs of distress, including prolonged withdrawal or hunched posture, prolonged expression of stereotypies, or excessive fearful behaviours e. Overseen by a dedicated individual with behavioural science or veterinary expertise f. Behavioural management strategies/procedures are documented in SOPs/protocols g. Behavioural management forms part of research protocol review	Bloomsmith, 2017; Bloomsmith et al., 2018; Schapiro, 2021
	0	Programme includes more than four of the above characteristics
	1	Programme includes two to four of the above characteristics
	2	Programme includes less than two of the above characteristics
11. Humane euthanasia programme Use 11a and 11b. 11a. Euthanasia programme	0	Personnel responsible for carrying out euthanasia are knowledgeable and competent to perform the procedure in a compassionate, professional, and appropriate manner that avoids distress to the animals; AVMA-approved methods are used	Canadian Council on Animal Care, 2019; Lambeth et al., 2013; American Veterinary Medical Association, 2020
	2	Personnel responsible for carrying out euthanasia are not suitably trained and competent; euthanasia methods are not AVMA-approved
11b. Humane endpoints		Characteristics of a high-quality programme: a. Accurate and clearly defined humane endpoints are established prior to study initiation b. Animals are monitored at an appropriate time and frequency to enable the earliest possible euthanasia decision and avoid moribundity/spontaneous death c. Humane endpoints are regularly reviewed and refined over time, as new data becomes available from studies using the model d. All personnel performing endpoint criteria assessments are able to recognise the signs of ill health e. Structured welfare assessment/humane endpoint score sheets are available and included in SOPs f. Where applicable, technologies such as telemetry, actimetry, imaging, and CCTV are used to inform endpoints decisions	Association of Primate Veterinarians, 2020; Prescott et al., 2021
	0	Programme includes more than four of the above characteristics
	1	Programme includes two to four of the above characteristics
	2	Programme includes less than two of the above characteristics
12. Health monitoring programme	0	Comprehensive health monitoring programme is in place, (e.g. as specified by FELASA), to track colony health and prevent disease outbreaks	Balansard et al., 2019
	2	No comprehensive health monitoring programme
13. Staff training Use 13a and 13b. 13a. Staff training progamme	0	All relevant staff (research, veterinary, and animal care) undergo a structured training programme before working with macaques; individual training records are kept and regularly reviewed (e.g. annually)	Wolfensohn & Honess, 2008; Jennings & Prescott, 2009
	1	A structure programme is in place, but records are not kept or regularly reviewed
	2	No structured programme in place
13b. Continuing professional development	0	All relevant staff (research, veterinary, and animal care) have the opportunity to attend internal and/or external presentations, conferences, and/or workshops on macaque welfare, care, and behaviour (e.g. NC3Rs Primate Welfare Meeting)
	2	Staff do not have access to continuing professional development opportunities
Sub-total: _ /16
Grand total: _ /45
Judgement: 0-15: Normal; assume good welfare state. 16-30: Welfare compromised; improvements required; monitor carefully. 31-45: Welfare severely compromised; suffering is likely; immediate action required; provide appropriate examination, treatment, and relief (e.g., analgesia, environmental adjustments).