Refinement

Inanimate Enrichment: Refinement

Reduction of aggression

Rodents
Ambrose N, Morton DB 2000. The use of cage enrichment to reduce male mouse aggression. Journal of Applied Animal Welfare Science 3, 117-125
"Aggression was significantly reduced by the addition of novel enrichment [cardboard box, softwood block, cardboard tube]. This reduction was expected as the provision of environmental enrichment for naive animals would decrease the familiarity of the cage and, therefore, lead to reductions of aggression. ... Even a simple enrichment aid such as a glass water bottle can significantly reduce postcage-cleaning aggression compared with mice kept in a barren cage."

Armstrong KR, Clark TR, Peterson MR 1998. Use of cornhusk nesting material to reduce aggression in caged mice. Contemporary Topics in Laboratory Animal Science 37(4), 64-66
The provision of cornhusk reduced aggressive interactions by offering subordinate animals cover and escape routes.

Arnold CE, Westbrook RD 1997/1998. Enrichment in group-housed laboratory golden hamsters. Animal Welfare Information Center (AWIC) Newsletter 8(3/4), 22-24
"Enriched hamsters [four same-sexed animals per group] showed varied behavior and less aggression toward their cagemates. The hamsters preferred jars to pipes" probably "because the jars' greater height, as compared to pipes, made it easy to look outside the cage."

*Cozens M 2006. Evaluation of the provision of hay to guinea pigs at GlaxoSmithKline. Animal Technology and Welfare 5, 31-32
"Two males in the group 3 had to be euthanized due to bite wounds from fighting with cage mates - this aggression is something we have also experienced in older male guinea-pigs where it is not possible to feed hay." Male groups who received hay were less difficult to handle and they were less aggressing among each other than animals who had no access to hay.

Gwinn LA, Krauthauser CL, Kerr JS 1999. Impact of home cage alterations on aggression in mice. Abstracts of the AALAS [American Association for Laboratory Animal Science] Meeting , 35 (Abstract)
PVC straight pipes, plumbing elbows and T pipes, and shreddible nesting squares were evaluated. "Nesting squares appear to be the most effective enrichment object for reducing the incidence of aggression in group-housed male mice."

Jones RB, Nowell NW 1973. The effects of familiar visual and olfactory cues on the aggression behaviour of mice. Physiology and Behavior 10, 221-223
Decreasing the familiarity of the environment and providing flexible avenues of escape lead to reduced levels of inter-male aggression. Greatest levels of aggression were found in cages that had ungergone incomplete cleaning, such as renewal of substrate only.

Marashi V, Barnekow A, Sachser N 2004. Effects of environmental enrichment on males of a docile inbred strain of mice. Physiology and Behavior 82, 765-776
"Environmental enrichment is intended to improve the welfare of laboratory animals. However, regarding male mice, numerous studies indicate an increase in aggressive behavior due to cage structuring. On the one hand, this might be a problem concerning animal welfare. On the other hand, enrichment is thought to hamper environmental standardization and to increase variability of data. ... From weaning until day 77+/-3 of life, animals [male mice] were kept in stable sibling groups of four under three different housing conditions: (A) nonstructured Makrolon type III laboratory cages ("standard housing"=S); (B) equivalent laboratory cages that were enriched with a box and scaffolding ("enriched housing"=E); and (C) spacious terrariums that were structured richly ("super-enriched housing"=SE). No differences in agonistic behavior, levels of plasma corticosterone (CORT), and activities of adrenal tyrosine hydroxylase (TH) existed among S-, E-, and SE-housed ABG males. Play behavior and general activity increased significantly with increasing enrichment. ...Enrichment did neither hamper standardization nor negatively influence the variability of physiological parameters. In summary, using a docile strain of mice revealed the positive effects of environmental enrichment also on male mice. The lack of adverse effects on behavior, physiology, standardization, and variability of data defuses these arguments against providing docile male mice with enrichment."

McClure DE, Thomson JI 1992. Cage enrichment for hamsters housed in suspended wire cages. Contemporary Topics in Laboratory Animal Science 31(4), 33 (Abstract)
"Golden Syrian hamsters (n=99) were housed individually in suspended wire cages so that spilled food and excreta could be removed. After 8 days, the hamsters developed bizarre aggressive behavior which consisted of growling, hissing, aggressive posturing toward humans, destruction of water bottle rubber stoppers, and attacking objects introduced into the cage. Many developed inappetence which progressed to anorexia, depression, and unresponsiveness. ... When cotton nestlets were provided to all of the hamsters, their appetite and responsiveness improved, but the aggressive behavior remained unchanged. The nestlets were replaced by a 13-cm length of 5.5-cm-diameter polyvinyl chloride pipe (PVC). The water bottles were replaced by an automatic watering system. After adding the PVC, the aggressive behavior diminished in 3 days and was unnoticeable in 14 days. In conclusion, when these hamsters were provided with nesting material their well-being was improved as indicated by resolution of inappetence and depression. Providing the PVC apparently resolved the aggressive behavior problem by providing a means for seclusion in addition to functioning as a burrow and as a toy."

Morrison P 2001. The rat floor pen: Fact or fantasy? Animal Technology 52, 33-34
"Aggression between rats is also decreased when they are housed in pens - no fights have been observed in a year. ... We feel that the floor pen offers many advantages for the animals besides increasing job satisfaction for the technicians."

Rabbits
Mis J, Warren F 2003. A novel and cost-effective approach to New Zealand White Rabbit enrichment. Tech Talk 8(6), 4
"A round children's pool is being used to serve as part of our enrichment program. .. We started our program by handling the [group-housed] rabbbits more frequently. . .We also noticed a dramatic decrease in rabbits showing aggressive behavior. .. They are a lot easier to handle not only for blood collection, but also when techniques such as nail clipping and routine exams are given."


Reduction of anxiety, fear and excitability

Rodents
Chapillon P, Manneché C, Belzung C, Caston J 1999. Rearing environmental enrichment in two inbred strains of mice: 1. Effects on emotional reactivity. Behavior Genetics 29, 41-46
Access to three times larger cages furnished with tunnels and running wheels reduced anxiety-like behavior.

Engellenner WJ, Goodlett CR, Burright RG, Donovick PJ 1982. Environmental enrichment and restriction: Effects on reactivity, exploration and maze learning in mice with septal lesions. Physiology and Behavior 29, 885-893
Mice from enriched living conditions showed less fear responses [resistance to capture, handling; vocalization] than mice from barren standard housing conditions .

Levine S 1985 A definition of stress? In Animal Stress. Moberg GP (ed), 51-69. Waverly Press, Baltimore, MD
Rats living in a more complex housing environment explored objects longer and more frequently than rats kept in barren standard cages, suggesting that enrichment buffers anxiety responses to potential stressors.

Manosevitz M, Pryor JB 1975. Cage size as a factor in environmental enrichment. Journal of Comparative and Physiological Psychology 89, 648-654
"Cage size significantly affected 38-day gross body weight, open-field activity and defecation, running-wheel activity, exploration, and water consumption. .. Rearing in large cages, either wire or Plexiglas, appears to reduce emotionality... The results of this experiment indicate that cage size is one of the important factors of the enriched-environment-rearing experience."

Meijer MK, Kramer K, Remie R, Spruijt BM, van Zutphen L. F. M., Baumans V 2002. Refinement of routine procedures: Can environmental enrichment reduce the stress response of laboratory mice. Fourth World Congress on Alternatives and Animal Use in the Life Sciences - Program and Abstracts , 97 (Abstract)
Female mice housed in trios in enriched cages and handled daily reacted with less anxiety to procedures than female mice housed singly in barren cages and handled only when necessary.

Morrison P 2001. The rat floor pen: Fact or fantasy? Animal Technology 52, 33-34
Rats "in [enriched] floor pens have better body condition and appear cleaner [than animals housed in cages]. They are also much more inquisitive and friendly than caged rats and come out when the technicians do their morning checks, because they appear less fearful of humans and want to interact with them."

Prior H, Sachser N 1994/95. Effects of enriched housing environment on the behaviour of young male and female mice in four exploratory tasks. Journal of Experimental Animal Science 37, 57-68
The provision of plastic boxes and wooden scaffoldings in cages reduced anxiety in mice, increased their open-field activity and improved sensorimotor skills.

Rabbits
Hansen LT, Berthelsen H 2000. The effect of environmental enrichment on the behaviour of caged rabbits (Oryctolagus cuniculus) . Applied Animal Behaviour Science 68, 163-178
Rabbits kept in conventional single-cages, showed more restlessness, excessive grooming, bar-gnawing and timidity than rabbits kept in cages that were provisioned with a platform and a shelter.

Reduction of stress

Rodents
Chamove AS 1989. Cage design reduces emotionality in mice. Laboratory Animals 23, 215-219
"The results of this study clearly showed that the presence of vertical partitions in the cages of mice reduced the stressfulness of caging and also led to mice which were less reactive when tested outside their cages. ... When reared in the more complex cages, the animals gained more weight and were more active."

Foulkes A 2004. Do laboratory rats benefit from more cage space? AWI Quarterly 53(3), 18-19
"Laboratory rats do not benefit from more cage space, unless the space is provisioned with proper enrichment such as a PVC tube. Being confined in a small standard cage is a much more stressful experience for rats than being confined in a larger cage furnished with a PVC tube. Access to a PVC tube reduces a rats stress level in a small cage, but the stress is still much higher than in a large cage with PVC tube." The animals stress status was reflected in their food consumption with rats living in barren cages eating much more food and gaining significantly more weight than rats living in the enriched cages. Rats kept in the typical, barren standard cages very often become obese. The present findings indicate that this problem is related to stressful living conditions, similarly to obesity in modern people living in stressful environments.


Hennesy MB, Foy T 1987. Non-edible material elicits chewing and reduces the plasma corticosterone response during novelty exposure in mice. Behavioral Neuroscience 101, 237-245
If mice were placed in a novel environment with metal foil or cardboard available, they chewed these materials. These animals had lower plasma corticosterone levels than mice in the same environment without access to these chewable items.

Van Loo PLP, Van der Meer E, Kruitwagen CLJJ, Koolhaas JM, Van Zutphen LFM, Baumans V 2004. Long-term effects of husbandry procedures on stress-related parameters in male mice of two strains. Laboratory Animals 38, 169-177
"Long-term provision of nesting material and its transfer during cage cleaning was found to influence several stress-related physiological parameters. Mice housed in cages enriched with nesting material had lower urine corticosterone levels and heavier thymuses, and they consumed less food and water than standard-housed mice. .. We conclude that the long-term provision of nesting material, including the transfer of nesting material during cage cleaning, reduces stress and thereby enhances the welfare of laboratory mice."

Reduction of abnormal behavior

Rodents

Callard MD, Bursten SN, Price EO 2000. Repetitive backflipping behaviour in captive roof rats (Rattus rattus) and the effect of cage enrichment. Animal Welfare 9, 139-152
"Repetitive stereotyped behaviours are often performed by both wild and domestic rodents in small laboratory cages. In this study, a behaviour resembling a backwards somersault or backflip is described and quantified in captive roof rats (ship or black rats, Rattus rattus). ... Cage enrichment in the form of a wooden nest box resulted in dramatically lower rates of performance. Increased cage height resulted in delayed development of backflipping, as well as changes in the form of the behaviour. Results are consistent with the hypothesis that the development and expression of backflipping in young roof rats may be triggered by weaning and maintained by a heightened state of arousal in a relatively impoverished environment with limited opportunities for perceptual and locomotor stimulation."

DeLuca AM 1997. Environmental enrichment: does it reduce barbering in mice? AWIC Newsletter 8(2), 7-8
"In order to alleviate barbering, their environment was enriched with toys and other objects at two different times. .. The toys were replaced every other week; at no time was the same toy used for more than 2 weeks. ... The extent of the barbering behavior was decreased by the enrichment program. If the environment was enriched as soon as the mice were gang-caged, barbering was kept at a minimum that is, only the smallest mice (less than 2 percent of the total number of animals) were barbered. If the enrichment was delayed for a month or more, barbering occurred to about 23 percent of the mice. Barbering in control mice (no enrichment) was about 60 percent."

*Gebhardt-Henrich SG, Vonlanthen. E.M., Steiger A 2005. How does the running wheel affect the behaviour and reproduction of golden hamsters kept as pets? Applied Animal Behaviour Science 95, 199-203
"Hamster females with a functional wheel showed significantly less climbing and stereotypical bar-mouthing than females with non-functional wheels."

*Hadley C, Hadley B, Ephraima S, Yang M, Lewis MH 2006. Spontaneous stereotypy and environmental enrichment in deer mice (Peromyscus maniculatus): Reversibility of experience. Applied Animal Behaviour Science 97, 312-322
"Enriched housing (24-84 day of age) was associated with substantially lower levels of stereotypy. Moreover, environmental enrichment experienced later in development (84-124 days of age) was as effective as early enrichment. This was not the case, however, for older animals (11-14 months of age) that experienced the same duration of enriched housing. The beneficial effects of early enrichment were found to persist as mice placed in standard cages following enrichment exhibited lower levels of stereotypy than controls. These findings support a sensitive period for the amelioration of abnormal repetitive behavior and suggest a neuroprotective effect of this form of early experience."

Orok-Edem E, Key D 1994. Responses of rats (Rattus norvegicus) to enrichment objects. Animal Technology 45, 25-30
" Two objects, a tongue depressor made of birch wood and a wooden block constructed of a stainless steel clip and cut broom handle were each introduced into two groups while the third group served as control. Aberrant behaviours like fighting and chewing of metal cage bars were drastically reduced."

Powell SB, Newman HA, McDonald TA, Bugenhagen P, Lewis MH 2000. Development of spontaneous stereotyped behavior in deer mice: effects of early and late exposure to a more complex environment. Developmental Psychobiology 37, 100-108
"We hypothesized that spontaneous stereotypies exhibited by deer mice housed under standard laboratory conditions were the result of environmental restriction and that a sensitive period exists for the development/prevention of stereotypies. Exposure to a more complex environment early in the post-weaning period resulted in substantially less stereotypy in the complex environment."

Waiblinger E, König B 2004. Refinement of gerbil housing and husbandry in the laboratory. ATLA (Alternatives to Laboratory Animals) 32(Supplement), 163-169
An artificial burrow system is described that prevents the development of stereotypic digging.

Waiblinger E 2002. Refinement of gerbil housing and husbandry in the laboratory. Fourth World Congress on Alternatives and Animal Use in the Life Sciences - Program and Abstracts , 101 (Abstract)
"Stereotypic digging was caused by the lack of a burrow .... stereotypic bar chewing was caused by premature separation of juveniles from their family before the birth of younger siblings."

Wiedenmayer C 1997. Stereotypies resulting from a deviation in the ontogenetic development of gerbils. Behavioural Processes 39, 215-221
The provision of an artificial external burrow reduces stereotypic digging.

Würbel H, Chapman R, Rutland C 1998. Effect of feed and environmental enrichment on development of stereotypic wire-gnawing in laboratory mice. Applied Animal Behaviour Science 60, 69-81
Enrichment significantly reduced stereotypic wire-gnawing in pair-housed male mice by 40%, presumably as a consequence of the cover provided by the cardboard tubes. This is substantiated by observations that the tubes were used as a place to retreat upon disturbance as well as for resting. As a consequence the animals showed more resting and less grooming in cages containing a cardboard tube-shelter.

Rabbits
Biczak M, Pellecchia D 2002. Environmental enrichment for rodents and rabbits. TriBranch Symposium 2002 , unpublished poster presentation
Provision of various toys and gnawing block resulted in a "significant reduction, or even elimination of hair loss and bar biting" in rabbits.

Hansen LT, Berthelsen H 2000. The effect of environmental enrichment on the behaviour of caged rabbits (Oryctolagus cuniculus) . Applied Animal Behaviour Science 68, 163-178
Rabbits kept in conventional single-cages, showed more restlessness, excessive grooming, bar-gnawing and timidity than rabbits kept in cages that were provisioned with a platform and a shelter.

Krohn TC, Ritskes-Hoitinga J, Svendsen P 1999. The effect of feeding and housing on the behaviour of the laboratory rabbit. Laboratory Animals 33, 101-107
"Feeding the animals at 14:00 h [wild rabbits forage primarily late in the afternoon and during the night!] reduced abnormal behaviour during the dark period compared to feeding at 08:00 h.

Moore S, Beeston D 1999. Rabbit caging developments at Zeneca. Animal Technology 50, 157-161
A well structured single-cage - including protruding front, hay rack and platform - and a floor-pen system are described. "The incidence of this [persistent tail biting] has been greatly reduced by introducing more enrichment items to the pens, but avoiding those that encourage confinement of the animal, i.e., long tubes/drainpipes."

Potter MP, Borkowski GL 1998. Apparent psychogenic polydipsia and secondary polyuria in laboratory-housed New Zealand White rabbits. Contemporary Topics in Laboratory Animal Science 37, 87-89
Three single-caged rabbits with psychogenic polydipsia [excessive drinking without apparent physiological reason] were given toys for cage enrichment, "and the abnormal behavior decreased in all three cases."

Brain function

Benefield AC, Greenough WT 1998. Effects of experience and environment on the developing and mature brain: Implications of laboratory animal housing. ILAR [Institute for Laboratory Animal Research] Journal 39(1), 5-11
"Immune system function appears to be enhanced in EC (complex environmental) rats relative to controls housed in standard cages."

Bennett EL, Diamond MC, Krech D, Rosenzweig MR 1964. Chemical and anatomical plasticity of brain. Science 146, 610-619
"The control animals are kept under colony conditions, housed three in a cage and exposed to ongoing activity in the room. ... For enhanced experience, animals were given Environmental Complexity and Training [groups of 10 to 12 animals in large cages provided with toys]. ... For the third condition - reduced experience - animals are caged singly in a dimly lit and quiet room where they cannot see or touch another animal. ... Our observations demonstrate that rats given enriched experience develop, in comparison to restricted littermates, greater weight and thickness of cortical tissue and an increase in total acetylcholinesterase activity of the cortex. ... These changes have been produced consistently in many replications ... they are found in adult as well as young animals. Control experiments have demonstrated that these changes cannot be attributed primarily to differential handling or locomotor activity. ... The demonstration of such changes merely helps to establish the fact that the brain is responsive to environmental pressure."

Guilarte TR, Toscano CD, McGlothan JL, Weaver SA 2002. Environmental enrichment: Beneficial effects in a rodent model of lead neurotoxicity. Fourth World Congress on Alternatives and Animal Use in the Life Sciences - Program and Abstracts , 236 (Abstract)
"Using a rodent model of lead-induced neurotoxicity, we show that environmental enrichment reverses cognitive and molecular deficits induced by this developmental neurotoxicant. .. At weaning, pups were removed from the lead exposure and housed in isolation or environmental enrichment cages (8 per cage). Enrichment cages were larger and contained toys, mazes, a running wheel, and a hammock. Rats exposed to lead and reared in an isolated environment were significantly impaired in acquiring spatial learning task. On the other hand, lead-exposed rats reared in an enriched environment perform as well as control enriched rats and better than control isolated rats. ... Findings suggest that the living environment is an important modifier on the effect of toxins on the central nervous system."

Kempermann G, Kuh HG, Gage FH 1997. More hippocampal neurons in adult mice living in an enriched environment. Nature 386, 493-495
Mice reared in an environment enriched with running wheels, tunnels and toys possess more hippocampal neurons than litter mates reared in barren standard cages. It is likely that these extra neurons contribute to the enhanced performance in learning tasks showed by enriched versus non-enriched mice.

Morley-Fletcher S, Rea M, Laviola G 2004. Environmental enrichment during adolescence reverses the effects of prenatal stress on anxiety-related behaviour and stress in rats. Animal Welfare 13(S), 251 (Abstract)
Environmental enrichment during adolescence reverses the effects of prenatal stress on anxiety-related behavior.

Tagney J 1973. Sleep patterns related to rearing rats in enriched and impoverished environments. Brain Research 53, 35-361
Rats housed in enriched versus barren cages show brain changes indicative of enhanced cerebral activity.

Learning performance

Cooper RM, Zubek JP 1958. Effects of enriched and restricted early environments on the learning ability of bright and dull rats. Canadian Journal of Psychology 12, 159-164
"The bright animals reared in enriched environments [group-housing with numerous enrichment gadgets] showed no improvement in learning ability over bright controls reared under normal laboratory condition [group-housing in barren cage]. The dull animals, on the other hand, benefited greatly from the enriched experience and attained a level of performance equal to that of the bright animals. Rearing in restricted environments had converse effects. The dull animals suffered no impairment as compared with dull controls, while the bright animals were retarded to the levels of the dulls in learning performance."

Molina-Hernández M, Téllez-Alcántara NP 2004. Rats socially-reared and full fed learned an autoshaping task, showing less levels of fear-like behaviour than fasted or singly-reared rats. Laboratory Animals 38, 236-245
"During the learning of instrumental tasks, rats are usually fasted to increase reinforced learning. However, fasting produces several undesirable side effects. The aim of this study was to test the hypothesis that control rats, i.e. full-fed and group-reared rats, will learn an autoshaping task to the same level as fasted or singly-reared rats. The interaction between fasting and single-rearing of rats was also tested. Results showed that control rats and fasted rats acquired the autoshaping task similarly, independently of rearing or gender. However, fasted or singly-reared rats produced fear-like behaviour. .. In conclusion, control rats learned the autoshaping task to the same level as fasted or singly-reared rats. However, fasting or single-rearing produced fear-like behaviour. Thus, the training of control rats in autoshaping tasks may be an option that improves animal welfare."

Patterson-Kane EG, Hunt M, Harper DN 1999. Behavioral indexes of poor welfare in laboratory rats. Journal of Applied Animal Welfare Science 2, 97-110
Problem solving ability was improved when rats were kept in groups in enriched cages rather than in barren single cages.

Immune function

Kingston SG, Hoffman-Goetz L 1996. Effect of environmental enrichment and housing density on immune system reactivity to acute exercise stress. Physiology and Behavior 60, 145-150
Keeping mice in enriched cages rather than barren standard cages seems to enhance immune function. "Environmentally enriched mice behave immunologically much like nonstressed animals." Animals in enriched environment showed a less variable, better regulated immune response.

Wound healing

Coviello-Mclaughlin GM, Starr SJ 1997. Rodent enrichment devices - evaluation of preference and efficacy. Contemporary Topics in Laboratory Animal Science 36(6), 66-68
"When animals wearing wound clips were exposed to the preferred enrichment, premature wound clip removal decreased, suggesting a positive effect of enrichment on the psychological well-being of surgically manipulated mice."

Prevention of obesity

Eskola S, Kaliste-Korhonen E 1998. Effects of cage type and gnawing blocks on weight gain, organ weights and open-field behaviour in Wistar rats. Scandinavian Journal of Laboratory Animal Science 25, 180-193
"Rats gnawed blocks about four time more in GFCs [grid floor cages without bedding] than in SBCs [solid bottom cages with bedding]. .. The presence of blocks in cages decreased the weight gain in both cage types."

Van Berkum LE 2000. Use of a feeder insert to reduce obesity in rats. AALAS [American Association for Laboratory Animal Science] 51st National Meeting Official Program , 125 (Abstract)
"By inserting a modified stainless steel plate into the feeder, area of exposed food is reduced, and may result in increased exploratory activity, which may lead to decrease in body weights and food consumption values (all while continuing to provide ad libitum access to food). .. Although body weights and food consumption were not significantly different for weeks 1-9, weeks 10-14 showed a trend towards lower body weights and food consumption in the treated groups. .. The study will be continued ."

Wrightson D, Dickson C 1999. Diet restriction through hopper design. Animal Technology 50, 45-46
Group-housed rats were induced to 'work' for their food by soldering metal plates over their food hoppers, so that only 3% of the original area remains available. It was felt that this method of food restriction was preferable to giving less food [to avoid obesity]. ... Rather than rapidly eating a reduced ration and feeling hungry for long periods, the rats worked harder for their food, which enabled them to burn more calories and eat throughout the day. This reduces the incidence of obesity and its associated disorders and also encourages more 'natural' behaviour patterns, both of which improve welfare."

Table of Contents




Social Enrichment: Refinement

Social buffer: Reduction of anxiety, fear and stress

Rodents
Conger JJ, Sawrey WL, Turrell ES 1957. An experimental investigation of the role of social experience in the production of gastric ulcers in hooded rats. American Psychologist 12, 410 (Abstract)
Rats in a chronic approach-avoidance conflict situation alone had significantly greater resultant ulceration than animals tested together.

Davitz JR, Mason DJ 1955. Socially facilitated reduction of a fear response in rats. Journal of Comparative and Physiological Psychology 48, 149-151
The presence of a conspecific mediates fear responses to a stressful situation.

Kaiser S, Kirtzeck M, Hornschuh G, Sachser N 2003. Sex specific difference in social support - a study in female guinea pigs. Physiology and Behavior 79, 297-303
"In female guinea pigs social support can be provided by social partners. In contrast to males, however, not only the bonding partner is able to reduce the female's stress responses, but also a familiar conspecific, though in a less effective way."

Latané B 1969. Gregariousness and fear in laboratory rats. Journal of Experimental Social Psychology 5, 61-69
In a novel open-field environment rats showed less signs of fear [number of fecal boluses excreted] when tested in pairs versus alone.

Latané B, Glass D 1968. Social and nonsocial attraction in rats. Journal of Personality and Social Psychology 9, 142-146
Single rats show a reduction of fear in a strange environment when another - though anesthetized - rat is present.

Patterson-Kane EG, Hunt M, Harper DN 1999. Behavioral indexes of poor welfare in laboratory rats. Journal of Applied Animal Welfare Science 2, 97-110
Fearfulness associated with single-housing was alleviated by pair-housing, and even further alleviated by group-housing in enriched cages. Problem solving ability was improved when rats were kept in groups in enriched cages rather than in barren single cages.

Sharp JL, Azar T, Lawson D 2003. Does cage size affect heart rate and blood pressure of male rats at rest or after procedures that induce stress-like responses? Contemporary Topics in Laboratory Animal Science 42(3), 8-12
"Close interaction between group-housed rats appears to be more important to stress reduction than is providing increased floor space per animal."

Sharp JL, Zammit TG, Azar TA, Lawson DM 2002. Stress-like responses to common procedures in male rats housed alone or with other rats. Contemporary Topics in Laboratory Animal Science 41(4), 8-14
"We concluded that under resting conditions, rats housed four per cage were less stressed than were rats housed alone, that common procedures induce noteworthy stress-like responses in male rats, and that the magnitude and duration of these responses are reduced by group housing."

Sharp JL, Zammit T, Azar TA, Lawson DM 2003. Stress-like responses to common procedures in individually and group-housed female rats. Contemporary Topics in Laboratory Animal Science 42(1), 9-18
"Procedure-induced arousal behaviors occurred in all housing groups after the acute husbandry and experimental procedures, but the occurrence of these behaviors was less frequent and of shorter duration in group-housed rats than rats housed alone. In light of these results, we conclude that under resting conditions group housed rats were somewhat less stressed than were rats housed alone. Further, we conclude that common procedures induce significant stress-like responses in female rats, and the magnitude and duration of these responses are reduced by group housing."

Taylor GT 1981. Fear and affiliation in domesticated male rats. Journal of Comparative and Physiological Psychology 95, 685-693
"Unfamiliar conspecifics were just as effective in allaying fear as familiar animals. Even the individually reared rats, unused to other rats, were less fearful with conspecifics than when they were stressed alone. These findings simply attest to the strength of the capability of conspecifics to reduce fear."

Group and pair housing: Alleviation of abnormal behavior

Rabbits
Chu L, Garner JP, Mench JA 2004. A behavioral comparison of New Zealand White rabbits (Oryctolagus cuniculus) housed individually or in pairs in conventional laboratory cages. Applied Animal Behaviour Science 85, 121-139
The beneficial effects of pair housing in decreasing abnormal behaviors and increasing locomotion suggest that pair housing should be considered as an alternative to individual housing for caged laboratory rabbits."

Held SDE, Turner RJ, Wootton RJ 2001. The behavioural repertoire of non-breeding group-housed female laboratory rabbits (Oryctolagus cuniculus). Animal Welfare 10, 437-443
"Trichophagy and stereotypic behaviors observed in singly caged rabbits were not observed in group-housed does."

Krohn TC, Ritskes-Hoitinga J, Svendsen P 1999. The effect of feeding and housing on the behaviour of the laboratory rabbit. Laboratory Animals 33, 101-107
"While the [individually housed] rabbits in cages spent 2-5% of the time performing abnormal behaviour like biting the bars or scratching the bottom of the cage, these activities were virtually absent in group-housed rabbits in floor pens."

Lehmann M 1987 Interference of a restricted environment - as found in battery cages - with normal behaviour of young fattening rabbits. In Rabbit Production Systems Including Welfare. Auxilia T (ed), 257-268. Official Publications of the European Communities, Luxembourg, Belgium
Single-caged rabbits were not able to hop in rabbit-typical fashion; they were less active but more restless [frequent activity changes] and showed more stereotypical gnawing of the wire cages than group-housed animals.

Metz JHM 1987 Behavioural problems of rabbits in cages. In Rabbit Production Systems Including Welfare. Auxilia T (ed), 221-230. Official Publications of the European Communities, Luxembourg, Belgium
Group-housed rabbits interacted with hay 11% of the 24-h day. Single-caged rabbits appeared to be more restless and showed more self-directed behavior than group-housed rabbits.

Table of Contents




Training to Cooperate: Refinement: Reduction of Stress

Rodents

Huang-Brown KM, Guhad FA 2002. Chocolate, an effective means of oral drug delivery in rats. Lab Animal 31(10), 34-36
"We trained the animals to smell the chocolate and develop a taste for it by holding the animal and placing the chocolate into its mouth using a blunt metal applicator (in our project we used a 14-gauge oral gavage needle). No esophageal contact, nor any placement more proximal than the oral cavity was necessary. We handled the rats gently to avoid association of chocolate with averse stimuli. To train the rats to expect the treatment, we opened and closed the cage before chocolate administration. .. After individual administration and return of the animal to the cage, the caregiver then offered drug-free chocolate as a "reward" at the front of the cage with the blunt end of the gavage needle to condition the animals to this manner of treatment. ...Results from this technique demonstrated appropriate levels of drug absorption. .. The animals do not require individual housing, enhancing their social environment and reducing space usage. .. Housing the animals used for this study in groups of three per cage allowed for easy identification of animals as they received their chocolate pellets. ...The chocolate vehicle is ideal for timed delivery or when a drug needs to be administered at a certain time of day. .. In this study, 3 of 57 rats (5%) failed to become accustomed to the chocolate even after the training period and had to be restrained for drug administration."

Shyu WC, Nightingale CH, Tsuji A, Quintiliani R 1987. Effect of stress on pharmacokinetics of amikacin and ticarcillin. Journal of Pharmaceutical Sciences 76, 265-266
Aberrant serum ticarcillin concentrations were measured when the "animals were simply picked out, put on a laboratory bench, and wrapped in a towel to restrain them" for blood collection. No aberration occurred in quasi trained rats who were "handled daily for 3-4 d prior to study and then held and petted by the investigator throughout the experimental period. No towel was needed to restrain these rats." In the amikacin group, 'trained' rats showed a much slower absorption process compared with conventionally handled rats. Alterations in renal flow due to the stress resulting from the traditional handling procedure may have affected pharmacokinetics.

Rabbits

Marr JM, Gnam EC, Calhoun J, Mader JT 1993. A non-stressful alternative to gastric gavage for oral administration of antibiotics in rabbits. Lab Animal 22(2), 47-49
"Generally, rabbits receive oral medication by gastric intubation, a method that frequently requires more than one technician, is time consuming, and places unnecessary stress on the animal." A very simple but effective training technique is described which ensures that rabbits voluntarily cooperate during oral drug administration. "We coated the tip of the syringe with sucrose sample. Inserting the syringe through the bars of the cage, we placed it in the animal's mouth and injected the sucrose solution slowly to allow the rabbit to taste and drink the fluid. We repeated the procedure three times a day for a total of 15 minutes per session, and within two days, 80% of the [10] animals voluntarily swallowed the fluid from the syringe. The [2] rabbits that did not seek out the syringe usually took it with only minimal encouragement. At the onset of the therapy, we substituted the antibiotic for the sucrose solution. .... We continued coating the tip of the syringe with sucrose granules throughout the therapy, apparently masking any unpleasant sensations produced by the antibiotic." Eight of the ten rabbits cooperated within two days. They "would stand with their paws on the front of the cages, protrude their faces from between the bars, and appear to beg for the syringe containing the antibiotic. " This non-stress method of "giving tosufloxacin was successful in producing the desired serum and bone concentrations."

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Variables

Single Housing, Solitary Confinement, Barren Cage & Understimulation

Rodents

Baenninger LP 1967. Comparison of behavioural development in socially isolated and grouped rats. Animal Behaviour 15, 312-323
Single-housed males showed reduced activity and a greater incidence of stereotypical tail manipulation [substitute social response?] and pawing with hind legs than males housed in groups.

Baer H 1971. Long-term isolation stress and its effects on drug response in rodents. Laboratory Animal Science 21, 341-349
"This review of the literature has illustrated that isolation stress, especially long-term isolation stress in rats and mice, can affect the growth, behavior, physiological condition, and response to a wide variety of drugs. ... Isolation stress in mice produces abnormal behavior (such as aggressiveness, excitability, and nervousness); changes in body weight; and organ weights; blood changes (leukopenia, eosinopenia); and frequently increased adrenal function over that of community housed mice."

Cambardella P, Greco AM, Sticchi R, Bellotti R, Di Renzo G 1994. Individual housing modulates daily rhythms of hypothalamic catecholaminergic system and circulating hormones in adult male rats. Chronobiology International 11, 213-221
Male rats housed individually had higher levels of corticosterone and prolactin than did those housed six per cage.

Chance MRA 1956. Environmental factors influencing gonadotrophin assay in the rat. Nature 177, 228-229
The variation in ovary weight (test response) was greater if the cages were small, if there was frequent disturbance by changing cages and cage-mates, and if the rats were caged either singly or in groups larger than two, with the floor area per rat roughly constant.

Fiala B, Snow FM, Greenough WT 1977. Impoverished rats weigh more than enriched rats because they eat more. Developmental Psychobiology 10, 537-541
Rats housed in barren environments consume more food due to boredom and are heavier than enriched rats.

Heath M 1999. Preliminary behaviour data for single and pair housed rats. Animal Technology 50, 47-48
"Singly housed rats spent most of the time in the back of the cage, while pairs spent most time by the food hoppers. This may have been exacerbated by the presence of humans, as pair housed rats were more confident overall. Activity patterns were very different between singly and pair housed rats, in term of both the total time spent doing different things and the time when specific activities occurred. In particular, single rats spent far less time playing than pair housed rats."

Hockly E, Cordery PM, Woodman B, Mahal. A, van Dellen. A, Blakemore C, Lewis CM, Hannan AJ, Bates GP 2002. Enrichment slows disease progression in R6/2 Huntington's disease mice. Annals of Neurology 51, 235-242
Housing mice in standard laboratory cages increased the rate of disease progression while housing them in enriched cages slowed the progress to a more 'normal' rate. "Mice housed in such deprived conditions are unlikely to prove a good model of disease, and the effects of enrichment represent a partial reversal of the deleterious effects of relative environmental impoverishment."

Hurst JL, Barnard CJ, Nevison CM, West CD 1997. Housing and welfare in laboratory rats: Welfare implications of isolation and social contact among caged males. Animal Welfare 6, 327-347
Singly housed males showed reduced activity and a greater incidence of self-directed behaviors, especially tail manipulation and self-grooming than males housed in small groups of three. They also performed "behaviors apparently related to escape or seeking social information.

Kaliste-Korhonen E, Eskola S, Rekilä T, Nevalainen T 1995. Effects of gnawing material, group size and cage level in rack on Wistar rats. Scandinavian Journal of Laboratory Animal Science 22, 291-299
"In open field, the animals living alone were less active and they moved slower from the peripheral to central area than the animals living in groups The singly housed rats showed in general more variation in behavioural measures than the animals with cage mates."

Mackintosh JH 1962. Effect of strain and group size on the response of mice to "sconal" anaesthesia. Nature 194, 1304
The variance of the response to a barbiturate anaesthetic was greater if male mice were caged either singly or in groups of eight than if they were caged in pairs.

*Meijer MK, Kramer K, Remie R, Spruijt BM, van Zutphen LFM, Baumans V 2006. The effect of routine experimental procedures on physiological paramters in mice kept under different husbandry conditions. Animal Welfare 15, 31-38
Single-housing in a non-enriched environment significantly elevated heart rate and blood pressure and recovery time following routine experimental procedures relative to social-housing in an enriched environment.

Pérez C, Canal JR, Dominguez E, Campillo JE, Guillén M 1997. Individual housing influences certain biochemical parameters in the rat. Laboratory Animals 31, 357-361
Individual as opposed to group-housing of female rats provoked variations in certain biochemical parameters [glucose, triglycerides, food intake]. It was concluded that this circumstance could make scientific data unreliable or even dubious.

Shaw DC, Gallagher RH 1984 Group or singly housed rats? In Standards in Laboratory Animal Management. The Universities Federation for Animal Welfare 65-70. The Universities Federation for Animal Welfare, Potters Bar, UK
"In both sexes singly housed animals showed a markedly lower survival rate than group-housed animals. This effect became apparent when the animals reached the age of about 80 weeks. No specific cause could be identified."

Sherwin CM, Olsson IAS 2004. Housing conditions affect self-administration of anxiolytic by laboratory mice. Animal Welfare 13, 33-38
"Standard cages for laboratory rodents may induce greater anxiety than enriched cages."

Späni D, Arras M, König B, Rülicke T 2003. Higher heart rate of laboratory mice housed individually vs in pairs. Laboratory Animals 37, 54-62
"Many studies have shown that housing mice individually over a long period significantly alters their physiology, but in most cases measurement has required human interference and restraint for sampling. Using a radio-telemetry system with implantable transmitters, we recorded heart rate (HR), motor activity (ACT) and body temperature (BT) of freely moving male mice (NMRI) housed either individually or in pairs with an ovarectomized female. .. Even after several weeks of habituation to the social conditions, HR was increased in mice housed individually compared with mice housed in pairs, although their measured ACT did not differ. .. Furthermore, individually-housed mice had more, but shorter, resting bouts, indicating disruption of the normal circadian sleep pattern. Enhanced HR in individually-housed mice does not necessarily indicate stress, but might be an important physiological indicator of discomfort. The fact that individual housing alters basic physiological parameters in laboratory mice highlights the need to control for housing-dependent variation, especially in experiments that are sensitive to changes in these parameters."

Valzelli L 1973. The "isolation syndrome" in mice. Psychopharmacologia 31, 3055-320
Socially isolated mice differ from group-housed mice not only behaviorally, but also in their immunological responses, hormone levels, brain neurochemistry, learning ability, pain thresholds, and sensitivity to drugs.

Rabbits

Bigler L,Lehmann M 1991 Schlußbericht über die Prüfung der Tiergerechtheit eines Festwandkäfigs für Hauskaninchen-Zibben. Bern, Switzerland: Bundesamt für Veterinärwesen
Single-housed young rabbits revealed inactivity atrophies as a result of modified behaviors and lack of exercise in small, conventional cages. The provision of functional areas (e.g., elevated shelf and dark area for withdrawal) can reduce the incidence of behavioral disorders.

Gunn D, Morton DB 1995. Inventory of the behaviour of New Zealand white rabbits in laboratory cages. Applied Animal Behaviour Science 45, 277-292
Detailed ethogram of singly caged rabbits. "The level of mobility (1.2%) reflected their inability to hop normally and, in addition, they were unable to sit up, rear fully or stretch out because of spatial restriction. All rabbits showed stereotyped activities (11%). .... Any gregarious animal chronically housed in isolation should be considered abnormal."

Lehmann M 1984 Beurteilung der Tiergerechtheit handelsüblicher Batteriekäfige für Mastkaninchen. Bern, Switzerland: Bundesamt für Veterinärwesen (Bericht)
Single-housed young rabbits revealed inactivity atrophies and poor bone growth as a result of modified behaviors and lack of exercise in small, conventional cages.

Loeffler K, Drescher B, Schulze G 1991. Einfluß unterschiedlicher Haltungsverfahren auf das Verhalten von Versuchs- und Fleischkaninchen [German text with English abstract]. Tierärztliche Umschau 46, 471-478
Rabbits kept individually in conventional wire-floored cages showed species-atypical patterns of locomotion and resting, and behavioral disorders which were absent in group-housed animals kept in floor pens. "The findings confirm that housing rabbits in cages is not to be recommended."

Wagner JL, Hackel DB, Samsell AG 1974. Spontaneous death in rabbits resulting from gastric trichobezoars. Laboratory Animal Science 24, 826-830
"Hairball formation in humans occurs most frequently in emotionally disturbed females below the age of 30. It is possible that individual caging of rabbits over long periods of time could affect their psychological health and bring about abnormal grooming habits, thus contributing to hairball formation."

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Stress From Laboratory Procedures: (Restraint & handling, blood collection, injection, etc.)

Balcombe JP, Barnard ND, Sandusky C 2004. Laboratory routines cause animal stress. Contemporary Topics in Laboratory Animal Science 43(6), 42-51
Based on a literature review the conclusion was drawn that "routine handling, venipuncture, and orogastic gavage lead to elevations of heart rate, blood pressure, and glucocorticoid concentrations that persist for 30 to 60 min or more following the event, suggesting that despite their routine use in laboratory studies, these procedures are acutely stressful for animals."

Barrett AM, Stockham MA 1996. The effect of housing conditions and simple experimental procedures upon corticosterone level in the plasma of rats. Journal of Endocrinology 26, 97-105
Routine procedures such as handling and weighing and i.p. injections of saline induce significant increases in plasma corticosterone concentrations wihich remain supernormal for at least 2 hr.

Berkey DL, Meeuwsen KW, Barney CC 1990. Measurements of core temperature in spontaneously hypertensive rats by radiotelemetry. American Journal of Physiology 258, R743-749
Handling and restraint can cause significant increases in core temperature.

Besch EL, Chou BJ 1971. Physiological responses to blood collection methods in rats. Proceedings of the Society for Experimental Biology and Medicine 138, 1019-1021
Plasma glucose levels are related to the length of handling time during blood collection.

*Bolam S 2005. Multiple housing of male CD-1 mice for toxicological studies. Animal Technology and Welfare 4, 86-87
"Aggression peaked around 10 weeks into the [10 months] study and was usually observed following dosing or cage cleaning, ceasing once human activities in the room were completed."

*Bonnichsen M, Dragsted N, Hansen AK 2005. The welfare impact of gavaging laboratory rats. Animal Welfare 14, 223-227
A significant correlation between heart rate and dosage was observed until 10 min after gavaging.

Briese E, Cabanac M 1991. Stress hyperthermia: Physiological arguments that it is fever. Physiology and Behavior 49, 1153-1157
Manual restraint increases body temperature in rats.

Guhad FA, Nevalainen T, Lang CM 2003. Fecal corticosterone: A noninvasive method of stress assessment in rats. AALAS [American Association for Laboratory Animal Science] 54th National Meeting Official Program , 93-94 (Abstract)
"Repeated mild restraint with bare hands or confinement in a plexiglas tube also increased fecal coritcosterone levels (p<0.05). Control animals, which only witnessed others being restrained, also showed a significant increase in fecal corticosterone levels (p<0.05)."

Kramer K, van de Weerd H, Mulder A, Van Heijningen C, Bauman V, Remie R, Voss H-P, Van Zutphen BFM 2004. Effect of conditioning on the increase of heart rate and body temperature provoked by handling in the mouse. ATLA (Alternatives to Laboratory Animals) 32(Supplement), 177-181
"Entering the animal room by the technician, without performing the handling and/or the acoustic stimulus increased HR significantly. . .. Handling further increased the HR. .. After repeatedly entering the animal room at a fixed time point (2:30pm), HR was significantly reduced after 12 days... After 2 weeks, the animals seem to habituate to this type of disturbance, with the exception of the first entrance in the morning (9:30am). Disturbance at this time point may cause more stress than later on."

Krulich L, Hefco E, Illner P, Read CB 1974. The effects of acute stress on the secretion of LH, FSH, prolactin and growth hormone in the normal male rat, with comments on their statistical evaluation. Neuroendocrinology 16, 291-311
Restraint stress is associated with a significant rise in plasma prolactin and LH levels.

*Mikkelsen LF, Dragsted N, Hansen AK, Ottesen JL 2005. The effect of training for long term restraining of rats evaluated by telemtry. ALTEX [Alternatives to Animal Experimentation] 22, 53 (Abstract)
Cardiovsacular stress responses were significant on all ten days of 3-hour tube restraint periods.

Moynihan J, Brenner G, Koota D, Breneman S, Cohen N, Ader R 1990. The effects of handling on antibody production, mitogen responses, spleen number, and lymphocyte subpopulations. Life Sciences 46, 1937-1944
The immune response was depressed when mice were picked up by the tail and held gently in the palm without restraint. "From the methodological point of view, handling of experimental animals may be a significant uncontrolled factor in many in vivo experiments."

Pitman DL, Ottenweller JE, Natelson BH 1988. Plasma corticosterone levels during repeated presentation of two intensities of restraint stress: Chronic stress and habituation. Physiology and Behavior 43, 47-55
Corticosterone levels of control rats increased when the animals witnessed stress reactions of conspecifics during forced restraint.

*Sharp J, Azar T, Lawson D 2005. Selective adaptation of male rats to repeated social encounters and experimental manipulations. Contemporary Topics in Laboratory Animal Science 44(2), :28-31
"We concluded that male Sprague-Dawley rats adapted quickly, albeit partially, to social encounters but did not adapt to repeated injections, suggesting selective adaptation depending on the type of stimulus used."

Tabata H, Kitamura T, Nagamatsu N 1998. Comparison of effects of restraint, cage transportation, anaesthesia and repeated bleeding on plasma glucose levels between mice and rats. Laboratory Animals 32, 143-148
"Mice showed a fairly consistent rise in levels of blood glucose if unaccustomed to [blood collection procedure] handling or if the cages were transported to an adjacent room." The same procedures when performed in rats "seemed to have small or no observable effect on levels of plasma glucose."

Tuli J, Smith JA, Morton DB 1995. Corticosterone, adrenal and spleen weight in mice after tail bleeding, and its effect on nearby animals. Laboratory Animals 29, 90-95
Subjects did not habituate to the stress associated with blood collection [tail bleeding].

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Stress from Husbandry Routine

Cage cleaning

Conn CA, Borer KT, Kluger MJ 1990. Body temperature rhythm and response to pyrogen in exercising and sedentary hamsters. Medicine and Science in Sports and Exercise 22, 636-642
Findings show that the cleaning of cages is one of the strongest stressors in husbandry routine for hamsters.

Duke JL, Zammit TG, Lawson DM 2001. The effects of routine cage-changing on cardiovascular and behavioral parameters in male Sprague-Dawley rats. Contemporary Topics in Laboratory Animal Science 40(1), 17-20
"Half of the rats were moved to clean cages with fresh wood-chip bedding; the other animals were left undisturbed. Systolic, diastolic, and mean arterial blood pressures; heart rate; and cage behavior (movement, rearing, grooming) increased promptly and significantly when animals were placed in clean cages. These cardiovascular and behavioral responses lasted for 45 to 60 min. Those animals not moved to clean cages but present in the animal room when this procedure was done did not show significant increases in blood pressure, heart rate, or activity. When rats were moved to clean cages that contained new bedding plus a small quantity of the soiled bedding from their previous cage, the cardiovascular and behavioral responses were similar to those of animals exposed to completely fresh bedding. ... We conclude that ordinary animal husbandry procedures such as moving rats to a clean cage can induce transient, but significant, cardiovascular and behavioral changes. Investigators and animal care staff should recognize that such routine procedures could confound experiments conducted shortly thereafter."

Gray S, Hurst JL 1995. The effects of cage cleaning on aggression within groups of male laboratory mice. Animal Behaviour 49, 821-816
Dominant animals tend to be more aggressive in an environment with familiar odor than in a strange environment. It was shown that complete cage cleaning - new cage and new substrate - can minimize aggression among male mice as compared with partial cage cleaning.

Kuhnen G 1999. The effect of cage size and enrichment on core temperature and febrile response of the golden hamster. Laboratory Animals 33, 221-227
"Cleaning of cage "caused results similar to chronic housing in small cages." For the comparison and assessment of results, the description of the housing conditions seems to be as essential as the description of the experimental procedure."

Saibaba P, Sales GD, Stodulski G, Hau J 1996. Behaviour of rats in their home cages: daytime variations and effects of routine husbandry procedures analysed by time sampling techniques. Laboratory Animals 30, 13-31
"The cleaning regime [cages cleaned out and fresh sawdust with shredded paper provided] appeared to have a greater effect on behaviour [increased defecation and activity] than did time of day and the effects of cleaning lasted for several hours after the procedure had been completed.".

Sharp JL, Zammit T, Azar TA 2003. Are "by-stander" female Sprague-Dawley rats affected by experimental procedures? Contemporary Topics in Laboratory Animal Science 42(1), 19-28
Witnessing a routine cage change induced significant increases in HR in rats which were equal to or greater than those induced by witnessing decapitations.

Van Loo PLP, Kruitwagen CLJJ, Van Zutphen LFM 2000. Modulation of aggression in male mice: Influence of cage cleaning regime and scent marks. Animal Welfare 9, 281-295
"Group housing of male laboratory mice often leads to welfare problems due to aggressive behaviour. ... Aggression peaks after disturbances such as cage cleaning .. Our results indicated that neither kinship nor distribution of urine marks affected aggression. Olfactory cues from nesting and bedding material, however, affected aggression to a marked degree: transfer of nesting material reduced aggression significantly, while transfer of sawdust containing urine and faeces seemed to intensify aggression. ... We conclude that the transfer of nesting material will reduce aggression, or at least slow down its development, and thus aid the reduction of social tension due to cage cleaning."

Transfer to unfamiliar environment, transportation

Damon EG, Eidson AF, Hobbs CH, Hahn FF 1986. Effect of acclimation to caging on nephrotoxic response of rats to uranium. Laboratory Animal Science 36, 24-27
It took rats a minimum of 3 to 5 days to behaviorally adjust [feeding/drinking] to being transferred to a different cage type.

Drozdowicz CK, Bowman TA, Webb ML, Lang CM 1990. Effect of in-house transport on murine plasma corticosterone concentration and blood lymphocyte population. American Journal of Veterinary Research 51, 1841-1846
"A significant increase in plasma corticosterone concentration was seen in mice immediately after [in-house] transport. The normal circadian rhythm of plasma corticosterone concentration was altered for the subsequent 24-hour period. .. The .. acute increase in plasma corticosterone concentration was associated with alterations in the cellular components of the immune system. Result of the study indicated that routine in-house transport of laboratory mice should be considered a stressful stimulus. .. Therefore, in-house transport of laboratory animals should be viewed as an experimental variable to be controlled by the investigator."

Duke JL, Zammit TG, Lawson DM 2001. The effects of routine cage-changing on cardiovascular and behavioral parameters in male Sprague-Dawley rats. Contemporary Topics in Laboratory Animal Science 40(1), 17-20
"Systolic, diastolic, and mean arterial blood pressures; heart rate; and cage behavior (movement, rearing, grooming) increased promptly and significantly when animals were placed in clean cages. These cardiovascular and behavioral responses lasted for 45 to 60 min."

Euker JS, Meites J, Riegle GD 1975. Effects of acute stress on serum LH and prolactin in intact, castrate and dexamethasone-treated male rats. Endocrinology 96, 85-92
Placing rats into transfer boxes and moving them to another room triggers a significant rise in plasma prolactin and LH.

Fenske M 1992. Body weight and water intake of guinea pigs: influence of single caging and an unfamiliar new room. Journal of Experimental Animal Science 35, 71-79
"No changes in body weight are observed if adult male guinea pigs are removed from their groups and kept singly in their home enclosure. In contrast, subjects markedly lose weight and show a marked reduction of water intake if isolated from their groups and caged singly in an unfamiliar test room."

Friedman SB, Ader R 1967. Adrenocortical response to novelty and noxious stimulation. Neuroendocrinology 2, 209-212
It was found in rats that the magnitude of the stress response to being transferred into an experimental cage is virtually the same as that produced by electrical shock. ... Exposure to a new environment, or novelty, may contribute significantly to the adrenocortical response often attributed to the effects of noxious or painful stimulation."

Gärtner K, Büttner D, Döhler R, Friedel J, Lindema J, Trautschold I 1980. Stress response of rats to handling and experimental procedures. Laboratory Animals 14, 267-274
Serum corticosterone levels increased significantly within four minutes, reached a peak at 15 minutes and were back to control values 60 minutes after cage displacement. Prolactin, TSH, LH and FSH serum concentrations were also affected by cage movement. "The results support the conclusion that rats are not disturbed by the presence of people with whom they are familiar, such as the usual animal attendant for that room, if they do not touch the cage."

Landi MS, Kreider JW, Lang CM 1982. Effects of shipping on the immune function in mice. American Journal of Veterinary Research 43, 1654-1657
Mice need at least 48 h after transportation by truck or aeroplane to recover in terms of corticosteroid levels.

Sharp JL, Azar TA, Lawson DM 2003. Selective adaptation of male rats to repeated social and experimental stimuli. AALAS [American Association for Laboratory Animal Science] 54th National Meeting Official Program, 126-127 (Abstract)
Persistently elevated blood pressure suggested that rats do not adapt to being transferred to another room. This observation "is similar to the blood pressure changes observed in conditioned fear responses."

Tabata H, Kitamura T, Nagamatsu N 1998. Comparison of effects of restraint, cage transportation, anaesthesia and repeated bleeding on plasma glucose levels between mice and rats. Laboratory Animals 32, 143-148
"Mice showed a fairly consistent rise in levels of blood glucose if unaccustomed to [blood collection procedure] handling or if the cages were transported to an adjacent room." The same procedures when performed in rats "seemed to have small or no observable effect on levels of plasma glucose."

Tuli J, Smith JA, Morton DB 1995b. Stress measurements in mice after transportation. Laboratory Animals 29, 132-138
Subjects showed stress responses [cortisol, behavior] to being moved in the familiar home cage from one room to another.

Ursin H, Murison R 1986. Facts, fiction and rational decisions. Acta Physiologica Scandinavica 554(Supplement 1), 234-242
Rats showed a significant increase in plasma glucocorticosteroids after being moved by lift from one room to another, even though they remained in their home cages.

York JL, Regan SG 1982. Conditioned and unconditioned influences on body temperature and ethanol hypothermia in laboratory rats. Pharmacology, Biochemistry and Behavior 17, 119-124
An increase in body temperature occurred in animals whose individual cages were removed from their usual position. After one hour at the new location body temperature had increased 1.3ºC. Movement of an entire cage rack, with the cages remaining in their usual shelf position, caused only a relatively small elevation in body temperature (about 0.3ºC) after one hour.

Young LA, Pavlovska-Teglia G, Stodulski G, Hau J 1996. Effect of group housing and oral corticosterone administration on weight gain and locomotor development in neonatal rats. Animal Welfare 5, 167-176
"The stress response of rats induced by transfer from communal housing to single housing prior to parturition has a negative influence of the offspring. This is manifested by slower growth rate and a retardation in locomotor development."

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Light

Bellhorn RW 1980. Lighting in the animal environment. Laboratory Animal Science 30, 440-450
"What we basically have done to date is to provide lighting suitable to our needs and assumed it was all right for the animal." Light intensities in stacked cages vary substantially.

Clough G 1982. Environmental effects on animals used in biomedical research. Biological Reviews 57, 487-523
"The intensity of light in animal cages is likely to be the most variable environmental factor in the average animal room."

Clough G, Donnelly HT 1983 Light intensity influences the oestrous cycle of LACA mice. In Standards in Laboratory Animal Management. The Universities Federation for Animal Welfare (ed), 60. The Universities Federation for Animal Welfare, Potters Bar, UK
The variation in light intensities in the center of multi-tier, translucent rodent cages can exceed an 8-fold difference at bottom rows from top rows.

Ross S, Nagy ZM, Kessler C, Scott JP 1966. Effects of illumination on wall-leaving behavior and activity in three inbred mouse strains. Journal of Comparative and Physiological Psychology 62, 338-340
"The results clearly indicated that illumination levels is an important variable affecting the general activity of mice." The animals showed less "fearfulness" and "emotionality" under low illumination conditions.

Weihe WH, Schidlow J, Strittmatter J 1969. The effect of light intensity on the breeding and development of rats and golden hamsters. International Journal of Biometeorology 13, 69-79
"It was noticed that animals subjected to an illumination of 2000 lux were tame and playful with handling, while those at lower light intensities resisted handling and tried to bite when vaginal smear was taken. ... The weight of some important organs, such as adrenals and testes, and also the breeding performance, showed a significant relationship to light intensity which was not seen in the hamster. ... For practical purposes it can be inferred, that, to obtain uniform results, rooms for rat breeding need to be more equally illuminated. ... The different light intensities from 1 to 5,000 lx, that we have found in animal rooms, may have some effect on the responses of animals to experimental procedures."

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Various

Exposure to stressed conspecific

Beynen AC 1992. Communication between rats of experiment-induced stress and its impact on experimental results. Animal Welfare 1, 153-159
Stressful experimental procedures can give rise to signals or odours that affect non-treated animals present in the same or adjacent room.

De Laat JMT, Van Tintelen G, Beynen AC 1989. Transportation of rats affects behavior on non-transported rats in the absence of physical contact. Zeitschrift für Versuchstierkunde 32, 235-237
Exposure [visual, auditory and olfactory] to stressed conspecifics was accompanied by an increase in cortisol levels.

Fuchs E, Fluegge G, Hutzelmeyer HD 1987. Response of rats to the presence of stressed conspecifics as a function of day time. Hormones and Behavior 21, 245-252
Corticosterone levels of non-handled rats increase when the animals witness stress reactions of conspecifics during forced restraint.

Guhad FA, Nevalainen T, Lang CM 2003. Fecal corticosterone: A noninvasive method of stress assessment in rats. AALAS [American Association for Laboratory Animal Science] 54th National Meeting Official Program , 93-94 (Abstract)
"Repeated mild restraint with bare hands or confinement in a plexiglas tube also increased fecal coritcosterone levels (p<0.05). Control animals, which only witnessed others being restrained, also showed a significant increase in fecal corticosterone levels (p<0.05)."

Iimori K, Tanka M, Kohno Y 1982. Psychological stress enhances noradrenaline turnover in specific brain regions in rats. Pharmacology Biochemistry and Behavior 16, 637-640
Exposure [visual, auditory and olfactory] to stressed conspecifics was accompanied by an increase in cortisol levels.

Mackay-Sim A, Laing DG 1980. Discrimination of odours from stressed rats by non-stressed rats. Physiology and Behavior 24, 699-704
Placing a rat into a new cage triggers release of corticosterones. This endocrine response is augmented if the new environment is one in which other rats had previously been subjected to stress.

Pitman DL, Ottenweller JE, Natelson BH 1988. Plasma corticosterone levels during repeated presentation of two intensities of restraint stress: Chronic stress and habituation. Physiology and Behavior 43, 47-55
Corticosterone levels of control rats increased when the animals witnessed stress reactions of conspecifics during forced restraint.

Sharp JL, Zammit TG, Lawson DM 2002. Stress-like responses to common procedures in rats: Effect of the estrous cycle. Contemporary Topics in Laboratory Animal Science 41(4), 15-22
"We conclude that cycling female rats show stress-like effects when they are subjected to or witness common husbandry and experimental procedures, but there is little to no effect of the estrous cycle."

Separation from companion

Ehlers CL, Kaneko WM, Owens MJ, Nemeroff CB 1993. Effects of gender and social isolation on electroencephalogram and neuroendocrine parameters in rats. Biological Psychiatry 33, 358-366
Separation resulted in significant sleep disturbances - as measured in EEG responses - and increase in HPA activity.

Lawson DM, Churchill PC 2000. The effects of enrichment on parameters in hypertensive rats . Contemporary Topics in Laboratory Animal Science 39(1), 9-13
"Separation from a cage mate clearly induced the greatest activation, causing sustained increases in systolic and diastolic pressure and heart rate."

Cage level & multi-tier system

Ader DN, Johnson SB, Huang SW, Riley WJ 1991. Group-size, cage shelf level, and emotionality in nonobese diabetic mice - Impact on onset and incidence of IDDM. Psychosomatic Medicine 53(3), 313-321
'Emotionality' or 'fearfulness' is higher in mice housed in steel cages at the top of the rack.

Kaliste-Korhonen E, Eskola S, Rekilä T, Nevalainen T 1995. Effects of gnawing material, group size and cage level in rack on Wistar rats. Scandinavian Journal of Laboratory Animal Science 22, 291-299
The animals living on the highest shelf of the rack differed from the others in their latency times of rearing and grooming."

Noise

Anthony A, Ackerman E, Lloyd JA 1959. Noise stress in laboratory rodents. I. Behavioral and endocrine response of mice, rats and guinea pigs. Journal of the Acoustical Society of America 31, 1430-1436
Noise affects the endocrine system.

Anthony A, Harclerode JE 1959. Noise stress in laboratory rodents. II: Effects of chronic noise exposure on sexual performance and reproductive function of guinea pigs. Journal of the Acoustical Society of America 31, 1437-1440 Chronic noise impacts reproductive performance in guinea pigs.

Barrett AM, Stockham MA 1996. The effect of housing conditions and simple experimental procedures upon corticosterone level in the plasma of rats. Journal of Endocrinology 26, 97-105
Deliberate exposure to noise resulted in a highly significant increase in the plasma corticosterone level of rats housed in groups of 20.

Color of cage

Sherwin CM, Glen EF 2003. Cage colour preferences and effects of home cage colour on anxiety in laboratory mice. Animal Behaviour 66, 1085-1092
'Home cage colour had a significant effect on body weight and food consumption as well as on behaviour in a raised plus maze."

Observer effect

Döhler KD, Gärtner K, Mühlen AV, Döhler U 1977. Activation of anterior pituitary, thyroid and adrenal glands in rats after disturbance stress. Acta Endocrinologica 86, 489-497
Cortisol levels increased within 5 minutes after the entry of an investigator and reached peak levels after 15 minutes.

Gärtner K, Büttner D, Döhler R, Friedel J, Lindema J, Trautschold I 1980. Stress response of rats to handling and experimental procedures. Laboratory Animals 14, 267-274
"The results support the conclusion that rats are not disturbed by the presence of people with whom they are familiar, such as the usual animal attendant for that room, if they do not touch the cage."

Kramer K, van de Weerd H, Mulder A, Van Heijningen C, Bauman V, Remie R, Voss H-P, Van Zutphen BFM 2004. Effect of conditioning on the increase of heart rate and body temperature provoked by handling in the mouse. ATLA (Alternatives to Laboratory Animals) 32(Supplement), 177-181
"It can be concluded that entering the animal room, even without handling of the animals, increases HR and BT. After 2 weeks, the animals seem to habituate to this type of disturbance, with the exception of the first entrance in the morning (9:30am). In order to mimic standard procedures as much as possible, the daynight cycle was not reversed. Thus, the first entrance is in the beginning of the animals rest period. Disturbance at this time point may cause more stress than later on. Repeated handling with or without conditioning by an acoustic stimulus reduces the increase in HR after 12 days especially in the afternoon. .. Thus, it can be concluded that conditioning of mice can be effective in attenuating the stress effects of routine procedures."

Environmental enrichment

Augustsson H, Van de Weerd HA, Kruitwagen CLJJ, Baumans V 2003. Effect of enrichment on variation and results in the light/dark test. Laboratory Animals 37, 328-251
"The aim of this study was to investigate if cage enrichment induces an effect on experimental mean values and on inter-individual variation in the light/dark paradigm using diazepam as the anxiolytic drug. The behaviour of 216 naive adult male mice of two different strains (BALB/c and C57BL/6) was studied. The animals were housed in groups of four in 'non-enriched', 'enriched' (nesting material) or 'super-enriched' (nest-box, nesting material, wooden gnawing stick and PVC tube) cages. .. Mice housed in enriched and super-enriched cages gained more weight than mice in non-enriched cages, although food intake was not affected. Generally, the strain of mouse had the greatest impact on both mean values and variation. .. None of the [statistical] methods showed any significant differences between standard and enriched conditions on variability in any of the parameters measured."

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Species-Typical Behavior

Rodents

Rats
Anne 2005. Norway Rat Behavior Repertoire - An informal description of rat behavior. Web site
Excellent description of species typical rat behavior.

Berdoy M 2003. The Laboratory Rat: A Natural History [a 27 min. video]. Berdoy, M., Oxford, UK
"Shot as a wildlife documentary over several months, this 27 min film follows the lives of domestic rats after being released in a large outdoor enclosure where they have to compete, like their wild cousins, for food, shelter and mates. When keeping animals in captivity, it is important to know what they have evolved to do. Progress in animal welfare is, to a large extent, driven by a combination of awareness, willingness and facts. This film aims to be relevant to all three by reviewing the range of behaviours and needs which, despite generations of domestification, remain innate and ready to be expressed when given the opportunity. The topics covered in the film include the following: 1. Food selection; 2. The benefits of living in groups; 3. Learning abilities; 4. Dominance hierarchies;5. Anti-predation; 6. Communication (olfactory and ultrasonic); 7. Habitat selection and orientation; 8. Mating strategies and sexual selection; 9. Neophobia; 10. Birth & Infanticide."

Jegstrup IM, Vestergaard R, Vach W, Ritskes-Hoitinga M 2005. Nest-building behaviour in male rats from three inbred strains: BN/HsdCpb, BDIX/OrIIco and LEW/Mol. Animal Welfare 14, 149-156
It was demonstrated in three inbred strains [BN, BDIX, LEW] of pair-housed male rats that naïve animals who never had access to any kind of nesting material do build nests if the correct stimuli are provided. The animals were housed in 48 x 38 x 20 cm large cages with aspen wood chips bedding, each furnished with a roofed wooden nest box measuring 28.5 x 20.5 x 12.5 cm. The box had two entrance holes with a 6 cm diameter on two different sides. Aspen wood wool and autoclaved straw were placed outside the box. The nest was removed and the box along with the two rats transferred to a new cage every two weeks. Additional fresh bedding material and straw was added twice a week, and wood wool once a week. The wooden nest box was machine-washed every two months. All animals built a new nest in the box in the two week period between the cage changing procedure. The animals began with the removal of all aspen bedding material form the nest box. They continued with the collection of straw and wood wool, forming a mat inside the empty nest box. This mat could be several centimeters thick and formed into a cup-shape nest. The actual construction of the nest consisted of fibers of both straw and wood wool, with straw forming a larger part of the nest structure itself.

Mice
Latham N, Mason G 2004. From house mouse to mouse house: the behavioural biology of free-living Mus musculus and its implications in the laboratory. Applied Animal Behaviour Science 86, 261-289
The species-typical behavior of free-living mice is described and analyzed in reference to its implication to the research laboratory setting.

Guinea Pigs
Kunkel P, Kunkel I 1964. Beiträge zur Ethologie des Hausmeerschweinchens Cavia aperea f. porcellus (L.) [Contribution to the ethology of the domesticated guinea pig (German text with English summary)]. Zeitschrift für Tierpsychologie 21, 602-641
Comprehensive report on the behavior of group-housed guinea pigs; with numerous photographs.

Reinhardt V 1971 Soziale Verhaltensweisen und soziale Rollen des Hausmeerschweinchens [Social behavior and social roles of guinea pigs]. Söcking/Starnberg, Germany: Dissertationsdruck Novotny
The social and individual behavior of heterosexually group-housed and isosexually pair-housed guinea pigs is described and analyzed. The role of olfactory and behavioral factors as social communicators and social buffers is underscored. The protective effect of submission is described and analyzed: If a male stops counter-aggression in a dispute with another male and yields without panic reaction, the victor will treat him like a female. Rather than showing submissive gestures, the subordinate partner will behave like an estrus female and stop emitting the male-typical pheromones. This trick effectively saves him from further attacks, and the dominant male will now treat him like a female and display courtship rather than aggressive behaviors. In fact, males are strictly inhibited from overt aggression against females, and the subordinate male who plays the female role runs no risk boxing the dominant opponent in typical female fashion. Running away stimulates rather than inhibits aggression if there is no place to hide.

Gerbils
Roper TJ, Polioudakis E 1977. The behaviour of Mongolian gerbils in a semi-natural environment with special reference to ventral marking, dominance and sociability. Behaviour 61, 207-237
The species-typical behavior of gerbils is described.

Rabbits

Kraft R 1978/79. Vergleichende Verhaltensstudien an Wild- und Hauskaninchen. I. Das Verhaltensinventar von Wild- und Hauskaninchen (pp. 140-162); II. Quantitative Beobachtungen zum Sozialverhalten (pp. 165-179) [German text with English summary]. Zeitschrift für Züchtungsbiologie 95, 140-162 & 165-179
Detailed, comparative description of rabbit-specific behavior of wild and domesticated animals.

Lehmann M 1991. Social behaviour of young domestic rabbits under semi-natural conditions. Applied Animal Behaviour Science 32, 269-292
Detailed description of species-typical behavior patterns.

Mykytowycz R 1958. Social behaviour of an experimental colony of wild rabbits (Oryctolagus cuniculus L.) 1. Establishment of the colony. CSIRO Wildlife Research 3, 7-25
Description of natural social behaviors and social relationships in rabbits.

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