Stress in cynomolgus monkeys (Macaca fascicularis) subjected to long-distance transport and simulated transport housing conditions

The stress associated with transportation of non-human primates used in scientific research is an important but almost unexplored part of laboratory animal husbandry. The procedures and routines concerning transport are not only important for the animals' physical health but also for their mental health as well. The transport stress in cynomolgus monkeys (Macaca fascicularis) was studied in two experiments. In Experiment 1, 25 adult female cynomolgus monkeys were divided into five groups of five animals each that received different diets during the transport phase of the experiment. All animals were transported in conventional single animal transport cages with no visual or tactile contact with conspecifics. The animals were transported by lorry for 24 h at ambient temperatures ranging between 20 degrees C and 35 degrees C. Urine produced before, during and after transport was collected and analysed for cortisol by enzyme-linked immunosorbent assay (ELISA). All monkeys exhibited a significant increase in cortisol excretion per time unit during the transport and on the first day following transport.Although anecdotal reports concerning diet during transport, including the provision of fruits and/or a tranquiliser, was thought likely to influence stress responses, these were not corrobated by the present study. In Experiment 2, behavioural data were collected from 18 cynomolgus macaques before and after transfer from group cages to either single or pair housing, and also before and after a simulated transport, in which the animals were housed in transport cages. The single housed monkeys were confined to single transport cages and the pair housed monkeys were kept in their pairs in double size cages. Both pair housed and singly housed monkeys showed clear behavioural signs of stress soon after their transfer out of their group cages.However, stress-associated behaviours were more prevalent in singly housed animals than in pair housed animals, and these behaviours persisted for a longer time after the simulated transport housing event than in the pair housed monkeys. Our data confirm that the transport of cynomolgus monkeys is stressful and suggest that it would be beneficial for the cynomolgus monkeys to be housed and transported in compatible pairs from the time they leave their group cages at the source country breeding facility until they arrive at their final laboratory destination in the country of use.     

Recommended housing conditions and test procedures can interact to obscure a significant experimental effect

Routine animal husbandry variables, such as group housing of mice and the order of testing of cagemates, are currently viewed to be essentially neutral with respect to the outcome of most, if not all, animal-based experiments, including those that utilize behavioral measurements. During the course of experiments that have utilized the elevated plus-maze to examine the ability of a bacterial challenge of mice to induce anxiety-like behavior, due to the activation of various cytokine pathways, we followed the recommendation of laboratory animal care staff to house the mice in pairs. When we tested the members of the pairs successively, it was found, for the first experimental set, that the behavior that reflects anxiety (time in closed arms) of the first-tested animal differed from that of the second-tested animal for both the experimental and the control animals and, critically, that these changes were in the opposite directions for the controls and the experimental animals, thus obscuring the effect of the experimental manipulation. A second, independent experimental set also obtained a significant effect for the order of testing effect in the bacterial-challenged group, but not in the saline control group, although a similar trend was evident in this group as well. These results indicate that special care should to be taken in implementing housing recommendations and that preliminary tests may be necessary to ensure that housing conditions do not interact with tests of the phenomenon under experimental investigation.     

Automated cognitive testing of monkeys in social groups yields results comparable to individual laboratory-based testing

Cognitive abilities likely evolved in response to specific environmental and social challenges and are therefore expected to be specialized for the life history of each species. Specialized cognitive abilities may be most readily engaged under conditions that approximate the natural environment of the species being studied. While naturalistic environments might therefore have advantages over laboratory settings for cognitive research, it is difficult to conduct certain types of cognitive tests in these settings. We implemented methods for automated cognitive testing of monkeys (Macaca mulatta) in large social groups (Field station) and compared the performance to that of laboratory-housed monkeys (Laboratory). The Field station animals shared access to four touch-screen computers in a large naturalistic social group. Each Field station subject had an RFID chip implanted in each arm for computerized identification and individualized assignment of cognitive tests. The Laboratory group was housed and tested in a typical laboratory setting, with individual access to testing computers in their home cages. Monkeys in both groups voluntarily participated at their own pace for food rewards. We evaluated performance in two visual psychophysics tests, a perceptual classification test, a transitive inference test, and a delayed matching-to-sample memory test. Despite the differences in housing, social environment, age, and sex, monkeys in the two groups performed similarly in all tests. Semi-free ranging monkeys living in complex social environments are therefore viable subjects for cognitive testing designed to take advantage of the unique affordances of naturalistic testing environments.     

Social and nonsocial behaviors of adult rhesus monkeys after amygdalectomy in infancy or adulthood.

At 6 yr of age six female rhesus monkeys that had sustained bilateral amygdalectomy in infancy, and five intact controls, were transferred to an observation cage where behaviors were recorded while the monkeys were (a) alone, (b) paired with unfamiliar stimulus animals, and (c) paired with familiar monkeys from the opposite experimental group. The five adult controls then underwent amygdalectomy, and all tests were repeated with the infant- and adult-operated animals. Infant-operated monkeys changed behaviors more rapidly than did intact controls in social and nonsocial situations, and their activity levels were less modified after a 24-hr period in the observation cage. They were subordinate to intact controls but expressed less fear than did controls when briefly placed with an unfamiliar aggressive animal. Adult amygdalectomy produced many changes in behavior, but these aberrations were identical to those observed in like-age monkeys that had been amygdalectomized in infancy. Infant-operated monkeys demonstrated more behavioral deficits at 6 yr than they had earlier in life.     

Taking personality selection bias seriously in animal cognition research: a case study in capuchin monkeys (Sapajus apella)

In most experimental work on animal cognition, researchers attempt to control for multiple interacting variables by training subjects prior to testing, allowing subjects to participate voluntarily, and providing subjects with food rewards. However, do such methods encourage selection bias from subjects’ personalities? In this study, we trained eighteen zoo-housed capuchin monkeys (Sapajus apella) for two experiments, under conditions of positive reinforcement (i.e. food rewards) and free-choice participation. Using a combination of behavioral and rater-based methods, we identified and validated five personality dimensions in these capuchins (Assertiveness, Openness, Neuroticism, Sociability, and Attentiveness). Scores on Openness were positively related to individual differences in monkey task participation, reflecting previous work showing that such individuals are often more active, curious, and willing to engage in testing. We also found a negative relationship between scores on Assertiveness and performance on tasks, which may reflect the trade-offs between speed and accuracy in these animals’ decision-making. Highly Assertive individuals (the most sociable within monkey groups) may also prioritize social interactions over engaging in research. Lastly, monkeys that consistently participated and performed well on both tasks showed significantly higher Openness and lower Assertiveness compared to others, mirroring relationships found between personality, participation, and performance among all participants. Participation and performance during training was clearly biased toward individuals with particular personalities (i.e. high Openness, low Assertiveness). Results are discussed in light of the need for careful interpretation of comparative data on animal cognition and the need for researchers to take personality selection bias more seriously.     

One-trial long-lasting food-aversion learning in wild Japanese monkeys (Macaca fuscata)

We examined how Japanese monkeys in the wild formed an aversion to food which had been paired with poison. Ten monkeys of various ages and both sexes were chosen as subjects from 105 members of the Shiga-A1 troop at Jigokudani in Shiga Heights in Japan. We gave almond nuts to each subject. Once a monkey ate 10-20 almond nuts, he was captured and moved into an injection cage. Seven experimental subjects were injected intravenously with cyclophosphamide (20 mg/kg). Three control subjects received the same treatment except that they were injected with physiological saline. About 1 hour later, all subjects were released into the troop. The tests for conditioned aversions were conducted during the next 2 days. In the tests, the experimental subjects would not eat almond nuts, while the control subjects showed no hesitation in eating them. Five of the seven experimental subjects retained perfectly the aversion to almond nuts in tests conducted 1 month and 3 months later. The one-trial long-lasting food-aversion learning shown by the wild Japanese monkeys is discussed in terms of their feeding strategy. These results also suggest that food-aversion conditioning has potential as a nonlethal method for controlling crop-raiding monkeys.     

Targets,tactics, and the open mouth face during play fighting in three species of primates

The play fighting of many mammals involves the nonserious use of behavior patterns derived from serious fighting. A major question of theoretical importance has been that of how, given this overlap in patterns of behavior, the animals can distinguish between playful and nonplayful intent. One proposed solution is that animals use play signals to inform each other about the playful intent of their actions. The most widely reported play signal amongst primates is the open mouth play face. The manner in which this so-called signal functions is based on correlational evidence, with most reports simply noting its presence or absence in a given species. This study involved a detailed video-based analysis of the occurrence of open mouths during the play fighting of three species of primates. One captive troop each of ring-tailed lemurs, black-handed spider monkeys, and patas monkeys was used. By examining all open mouths in the context of the species-typical style of play fighting, several conclusions were empirically verified. 1) Most open mouths occur as a functionally necessary precursor for biting. 2) Some open mouths occur as a defensive threat which deters further contact. 3) The residual open mouths which may function as contact promoting play signals, constituted about 20–25% of all open mouths by the lemurs and patas monkeys, but less than 5% for spider monkeys. These species differences appeared to arise from two causes. Firstly, the spider monkeys used another signal, the head shake, in situations where lemurs and patas monkeys used open mouths. Secondly, the style of play fighting greatly influenced the frequency and duration of open mouths. This was most marked in the face-to-face combat style of patas monkeys. These findings show that comparative studies of the occurrence and function of play signals need to take into account species-typical styles of playful combat. Aggr. Behav. 23:41–57, 1997. © 1997 Wiley-Liss, Inc.     

Autonomic concomitants of discriminative avoidance and punishment training in the monkey

TwoCebus albifrons monkeys were trained to press a back-lighted panel to postpone a brief electric shock to the tail using a Sidman avoidance schedule (SS = 40 sec, RS = 40 sec). After 25 training sessions, a discriminative schedule was introduced, with the Sidman avoidance continuing in the presence of one discriminative stimulus and punishment introduced in the presence of the other. The discriminative stimuli were colors on the panel. Discriminative training also involved 25 sessions, each with a random sequence of 6 avoidance and 6 punishment segments, with 30 sec intervals between the segments. Plantar skin conductance and heart rate were recorded along with the panel-pressing behavior. The two monkeys adjusted to the discriminative schedule quite differently from one another. One animal responded at a high level and avoided very well (during avoidance) but was punished frequently (during punishment). The other animal responded less frequently and received many shocks during avoidance but almost none during punishment. The animal that showed less ability to inhibit responding (and received about four times as many shocks overall) appeared to have discriminated better temporally in spacing its responses during avoidance training. The monkey whose panel-pressing behavior resulted in more shocks also tended to show a higher tonic level of autonomic arousal. However, within-animal differences in shock frequency (between avoidance and punishment) were not similarly related to autonomie arousal. The animal that received fewer shocks overall (but more during avoidance) showed greater arousal during punishment. The animal that received more shocks overall (but fewer during avoidance) showed no arousal differences between avoidance and punishment.     

A cage-based training,cognitive testing and enrichment system optimized for rhesus macaques in neuroscience research

In neurophysiological studies with awake non-human primates (NHP), it is typically necessary to train the animals over a prolonged period of time on a behavioral paradigm before the actual data collection takes place. Rhesus monkeys (Macaca mulatta) are the most widely used primate animal models in system neuroscience. Inspired by existing joystick- or touch-screen-based systems designed for a variety of monkey species, we built and successfully employed a stand-alone cage-based training and testing system for rhesus monkeys (eXperimental Behavioral Intrument, XBI). The XBI is mobile and easy to handle by both experts and non-experts; animals can work with only minimal physical restraints, yet the ergonomic design successfully encourages stereotypical postures with a consistent positioning of the head relative to the screen. The XBI allows computer-controlled training of the monkeys with a large variety of behavioral tasks and reward protocols typically used in systems and cognitive neuroscience research.     

Macaques’ (Macaca mulatta) use of numerical cues in maze trials

We tested the ability of number-trained rhesus monkeys to use Arabic numeral cues to discriminate between different series of maze trials and anticipate the final trial in each series. The monkeys prior experience with numerals also allowed us to investigate spontaneous transfer between series. A total of four monkeys were tested in two experiments. In both experiments, the monkeys were trained on a computerized task consisting of three reinforced maze trials followed by one nonreinforced trial. The goal of the maze was an Arabic numeral 3, which corresponded to the number of reinforced maze trials in the series. In experiment 1 (n=2), the monkeys were given probe trials of the numerals 2 and 4 and in experiment 2 (n=2), they were given probe trials of the numerals 2–8. The monkeys receiving the probe trials 2 and 4 showed some generalization to the new numerals and developed a pattern of performing more slowly on the nonreinforced trial than the reinforced trial before it for most series, indicating the use of the changing numeral cues to anticipate the nonreinforced trial. The monkeys receiving probe trials of the numerals 2–8 did not predict precisely when the nonreinforced trial would occur in each series, but they did incorporate the changing numerals into their strategy for performing the task. This study provides the first evidence that number-trained monkeys can use Arabic numerals to perform a task involving sequential presentations.     

Control of responding by sound location in monkeys: rapid acquisition in darkness.

Rapid control of responding by sound location is obtained in squirrel monkeys when sound stimuli are presented from one of two loudspeakers, each one adjacent to a response key. With this arrangement of loudspeakers and response keys, squirrel monkeys quickly learn to respond on the key near the source of the sound stimulus, and this pattern is the same whether or not responses near the sound source are differentially reinforcedmthis result may depend on a pre-experimental tendency in squirrel monkeys to orient head and eyes toward a sound, which would lead the animal to look at the response key in front of the loudspeaker producing the sound. The present experiment sought to determine whether visual stimuli are necessary for rapid control of responding by sound location. Two monkeys were trained in darkness in a sound-localization task similar to that described above. Results were similar to those obtained from animals trained in light, indicating that visual stimuli are not required for rapid acquisition of sound-localization behavior in monkeys.     

Observational learning from tool using models by human-reared and mother-reared capuchin monkeys (<Emphasis Type="Italic">Cebus apella</Emphasis>)

Studies of wild capuchins suggest an important role for social learning, but experiments with captive subjects have generally not supported this. Here we report social learning in two quite different populations of capuchin monkeys (Cebus apella). In experiment 1, human-raised monkeys observed a familiar human model open a foraging box using a tool in one of two alternative ways: levering versus poking. In experiment 2, mother-raised monkeys viewed similar techniques demonstrated by monkey models. A control group in each population saw no model. In both experiments, independent coders detected which technique experimental subjects had seen, thus confirming social learning. Further analyses examined fidelity of copying at three levels of resolution. The human-raised monkeys exhibited fidelity at the highest level, the specific tool use technique witnessed. The lever technique was seen only in monkeys exposed to a levering model, by contrast with controls and those witnessing poke. Mother-reared monkeys instead typically ignored the tool and exhibited fidelity at a lower level, tending only to re-create whichever result the model had achieved by either levering or poking. Nevertheless this level of social learning was associated with significantly greater levels of success in monkeys witnessing a model than in controls, an effect absent in the human-reared population. Results in both populations are consistent with a process of canalization of the repertoire in the direction of the approach witnessed, producing a narrower, socially shaped behavioural profile than among controls who saw no model. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Spotting animals in natural scenes: efficiency of humans and monkeys at very low contrasts

The ability of monkeys to categorize objects in visual stimuli such as natural scenes might rely on sets of low-level visual cues without any underlying conceptual abilities. Using a go/no-go rapid animal/non-animal categorization task with briefly flashed achromatic natural scenes, we show that both human and monkey performance is very robust to large variations of stimulus luminance and contrast. When mean luminance was increased or decreased by 25–50%, accuracy and speed impairments were small. The largest impairment was found at the highest luminance value with monkeys being mainly impaired in accuracy (drop of 6% correct vs. <1.5% in humans), whereas humans were mainly impaired in reaction time (20 ms increase in median reaction time vs. 4 ms in monkeys). Contrast reductions induced a large deterioration of image definition, but performance was again remarkably robust. Subjects scored well above chance level, even when the contrast was only 12% of the original photographs (≈81% correct in monkeys; ≈79% correct in humans). Accuracy decreased with contrast reduction but only reached chance level -in both species- for the most extreme condition, when only 3% of the original contrast remained. A progressive reaction time increase was observed that reached 72 ms in monkeys and 66 ms in humans. These results demonstrate the remarkable robustness of the primate visual system in processing objects in natural scenes with large random variations in luminance and contrast. They illustrate the similarity with which performance is impaired in monkeys and humans with such stimulus manipulations. They finally show that in an animal categorization task, the performance of both monkeys and humans is largely independent of cues relying on global luminance or the fine definition of stimuli.     

A versatile dispenser for commercial monkey chow reinforcement

A dispenser is described that can deliver a wide variety of foodstuffs of almost any consistency. Despite compact size, storage capacity is sufficient for several days of unattended operation. Computer-compatible logic allows flexible applications in animal housing facilities and experimental setups. Its application in delivering regular monkey chow as reinforcement in experiments is discussed.     

Recognizing facial cues: individual discrimination by chimpanzees (Pan troglodytes) and rhesus monkeys (Macaca mulatta)

Faces are one of the most salient classes of stimuli involved in social communication. Three experiments compared face-recognition abilities in chimpanzees (Pan troglodytes) and rhesus monkeys (Macaca mulatta). In the face-matching task, the chimpanzees matched identical photographs of conspecifics' faces on Trial 1, and the rhesus monkeys did the same after 4 generalization trials. In the individual-recognition task, the chimpanzees matched 2 different photographs of the same individual after 2 trials, and the rhesus monkeys generalized in fewer than 6 trials. The feature-masking task showed that the eyes were the most important cue for individual recognition. Thus, chimpanzees and rhesus monkeys are able to use facial cues to discriminate unfamiliar conspecifics. Although the rhesus monkeys required many trials to learn the tasks, this is not evidence that faces are not as important social stimuli for them as for the chimpanzees.     

Primate head and body restraint without chronic skin openings or attachments to the animal

We developed a primate restraint system that requires no chronic skin openings or attachments to the animal. The restraining chair has a unique neck clasp; monkeys without chains and collars are easily trained to readily enter the chair and accept restraint with the neck and head held at a comfortable angle. A bite bar, in combination with contact on broad areas of the monkey’s brow and occiput, provides rigid head immobilization. In order to achieve contact with a broad area of the occipital bone, the muscles at the back of the animal’s head are surgically detached from the occiput and reattached to the underlying neck muscles. A strain-gauge, mounted on the head-holder and monitored by a laboratory computer, detects head movements of the monkey and permits the experimenter to teach the monkey to sit still during data acquisition. This system is well accepted by experienced monkeys and helps prevent the risks of infection posed by most earlier methods. Furthermore, the head and shoulders of the monkey are readily accessible for examination and for close positioning of test equipment.     

恒河猴数相加能力的初步研究

本实验训练恒河猴对两组其和不超过8的刺激实体(卡片上的黑色圆点)进行相加,再与第三组刺激实体进行数多少的比较判断。实验共有9个训练程序,课题逐次由易到难,刺激的数目也由少到多。有一只恒河猴达到了全部9个程序的训练标准。证明恒河猴的确具有学会把两个数进行相加的能力。这种相加不可能是数数的结果,极可能是在知觉上将它们融合或接合在一起。     

Anticipation of future events in squirrel monkeys (Saimiri sciureus) and rats (Rattus norvegicus): tests of the Bischof-Kohler hypothesis

The Bischof-Kohler hypothesis holds that nonhuman animals cannot anticipate a future event and take appropriate action when that event involves satisfaction of a need not currently experienced. Tests of the Bischof-Kohler hypothesis were performed with squirrel monkeys (Saimiri sciureus) and rats (Rattus norvegicus). In experimental trials with both species, a nonthirsty animal had its water bottle removed and then chose between a smaller and larger quantity of food. Consumption of the food induced thirst. Choice of the smaller quantity led to the return of the water bottle sooner than choice of the larger quantity. Monkeys reversed their baseline preference for the larger quantity of food when the experimental contingencies were introduced, but rats continued to prefer the larger amount. Although the rat findings support the Bischof-Kohler hypothesis, the monkey findings challenge it.     

Stock optimizing in choice when a token deposit is the operant.

Each of 2 monkeys typically earned their daily food ration by depositing tokens in one of two slots. Tokens deposited in one slot dropped into a bin where they were kept (token kept). Deposits to a second slot dropped into a bin where they could be obtained again (token returned). In Experiment 1, a fixed-ratio (FR) 5 schedule that provided two food pellets was associated with each slot. Both monkeys preferred the token-returned slot. In Experiment 2, both subjects chose between unequal FR schedules with the token-returned slot always associated with the leaner schedule. When the FRs were 2 versus 3 and 2 versus 6, preferences were maintained for the token-returned slot; however, when the ratios were 2 versus 12, preference shifted to the token-kept slot. In Experiment 3, both monkeys chose between equal-valued concurrent variable-interval variable-interval schedules. Both monkeys preferred the slot that returned tokens. In Experiment 4, both monkeys chose between FRs that typically differed in size by a factor of 10. Both monkeys preferred the FR schedule that provided more food per trial. These data show that monkeys will choose so as to increase the number of reinforcers earned (stock optimizing) even when this preference reduces the rate of reinforcement (all reinforcers divided by session time).     

Discrimination by rhesus monkeys of diets containing supplemental amino acids.

Following an opportunity to demonstrate a preference for water with or without the addition of the amino acid isoleucine, methionine, phenylalanine, or tryptophan at a concentration proportional to that in whole egg protein, 9 monkeys were subjected on 4 occasions to a 7-day experimental week when they received an isocaloric diet containing only one-fourth the amount of protein of their normal diet. An identical low-protein diet supplemented with one of the above amino acids, again at a concentration proportional to that in egg, was presented for an equivalent period during the experimental week and the amounts consumed of each diet were compared. Ss failed to exhibit a preference or an aversion for water supplemented with any of the amino acids; however, all low-protein diets supplemented with an amino acid were consumed in greater quantities than a low-protein diet lacking a supplement. On Days 6 and 7 of the experimental weeks when protein depletion was most severe, Ss significantly (p less than .05) preferred the diet supplemented with isoleucine to a diet lacking the supplement.