Successful implementation of cooperative handling eliminates the need for restraint in a complex non-human primate disease model

Background Streptozotocin‐induced diabetic non‐human primates are used to study efficacy and safety of innovative immunosuppression after islet transplantation. We implemented a training program for medical management of a chronic disease state. Methods Cooperation with hand feeding and drinking, shifting, and limb presentation were trained utilizing predominately positive but also negative reinforcement in 52 animals compared with 28 macaques subjected to conventional physical and/or chemical restraint. The success and timing of behavior acquisition was evaluated in a representative subset of 14 animals. Results Over 90% of animals were successful in behavior acquisition. Programmatically this resulted in complete elimination of chair restraint and negligible requirement for sedation. About half of the trained animals had no‐to‐moderate thymic involution, indicative of a substantial reduction in stress. Conclusion Cooperative handling enhances animal well‐being. This contributes to validity of scientific results and eliminates model‐induced confounding that can obstruct interpretation of safety and efficacy data.     

Effects of chair restraint on plasma enzyme values in the rhesus monkey (Macaca mulatta)

The purpose of this paper was to examine the effect of chair restraint on plasma enzyme values in the rhesus monkey. Six monkeys were restrained to the monkey chair for eight hours. Creatine phosphokinase (CK) value increased significantly three hours after the onset of restraint and LDH value did eight hours after the onset of restraint. The increase in CK, GOT and GPT values continued for 1 or 2 days after the release from restraint. On the other hand, these plasma enzyme values in non-restraint monkeys showed almost no changes. These results indicate that it is necessary to establish a proper method for the adjustment to chair restraint in the rhesus monkey.     

Effects of repeated chair restraint on physiological values in the rhesus monkey (Macaca mulatta)

The purpose of this study was to examine the effects of repeated chair restraint on some physiological values in the rhesus monkey. Although heart rate and blood pressure increased significantly at the onset of chair restraint, these increased values gradually decreased with repetition of restraint as well as with the lapse of time after restraining. Finally, no elevation of these levels was detected after repetitions lasting eight hours per day for five days. Plasma cortisol, CK and GOT values also showed similar trends. These elevated physiological values at the onset of chair restraint seemed to be due to high sympathetic tone and muscular activity used to try to avoid the restraint. It is concluded that repeated chair restraint has an accommodative component in monkeys resulting in the return of some elevated physiological values to the normal level.     

A Flexible Restraint Chair for the Cynomolgus Monkey (Macaca fascicularis)1

We have found that commercial restraint chairs suitable for the rhesus monkey (Macaca mulatta) cause severe physical distress to many cynomolgus monkeys (Macaca fascicularis). A new restraint chair has therefore been designed specifically for the cynomolgus. The key features of the chair are an angled neck piece, a large opening to provide free movement for the tail, a soft waist restraint, and a level surface for both the heels and ischial pads. This new chair is not only suitable for the cynomolgus monkey but may also be more comfortable for the rhesus monkey than standard commercial chairs.     

Training of laboratory-housed non-human primates in the UK

Abstract

Laboratory-housed non-human primates may experience a range of potential stressors, including physical and chemical restraint, venepuncture, injection, catching and cage-change. Training them to co-operate, using positive reinforcement techniques, is one means of significantly reducing the adverse impact of such procedures upon them and, therefore, is a refinement. Furthermore, the additional time that staff spend with the primates, and the need for individual recognition and close observation of animal behavior, mean that the trainer develops a relationship with each individual animal which can be beneficial for animal welfare (e.g., by reducing the occurrence of abnormal behavior and fear of humans). We surveyed use of training in thirteen UK establishments using and breeding primates, utilizing a mixed-mode questionnaire. The survey demonstrated that there is widespread awareness of training as a refinement and appreciation of its diverse benefits, but training is not used as widely or as fully as it might be. This is largely due to real constraints (principally staff and time and a lack of confidence in ability to train), and perceived constraints (such as a supposed lack of information on how to train and assessment of the benefits, and an overestimation of the time investment needed). There is also considerable variation between establishments in the purposes of training and techniques used, with a reliance on negative reinforcement in three. We conclude that there is considerable scope for refinement of common scientific, veterinary and husbandry procedures, and refer to some resources designed to help establishments take action (e.g., Prescott and Buchanan-Smith 2003).     

Alteration of high density lipoprotein subfraction distribution with induction of serum amyloid A protein (SAA) in the nonhuman primate

Overnight chair restraint results in a dramatic increase in serum amyloid A protein (apoSAA) of nonhuman primate high density lipoprotein (HDL). To determine whether apoSAA induction resulted in a displacement of indigenous HDL protein or a change in the subfraction distribution of HDL, we analyzed the characteristics of HDL subfractions in eight vervet monkeys before and 24 hr after apoSAA induction. Blood was taken from each animal before and after chair restraint to induce apoSAA. HDL was isolated from the plasma by ultracentrifugation and agarose column chromatography. The isolated HDL was subfractionated by density gradient centrifugation and five resulting subfractions were analyzed for protein and lipid content. With apoSAA induction there was a significant increase in d less than 1.09 g/ml protein, phospholipid, and free and esterified cholesterol which resulted in a 44% increase in the total mass of this subfraction. Concomitantly, there was a significant decrease in d 1.10-1.11 g/ml protein, total cholesterol, and cholesteryl ester, which resulted in a 16% decrease in the total mass of the subfraction. The response of the d 1.10-1.11 and d greater than 1.12 g/ml subfraction protein, cholesterol, and phospholipid concentrations to chair restraint for individual animals was directly proportional to their plasma HDL concentrations. Although there was a change in the HDL subfraction concentrations after chair restraint, there was no change in the lipid composition of the HDL subfractions nor in the total amount of HDL protein. However, the apoSAA/A-I ratio was significantly increased with induction while the apoA-II + C's/A-I ratio remained unchanged. The apoSAA/A-I ratio progressively increased with the density of the HDL subfraction. The protein composition of the d greater than 1.12 g/ml subfraction was changed from an average of three apoA-I and two apoA-II (or C's) molecules per particle to an average of two apoA-I, one apoA-II (or C's), and three or four apoSAA molecules per particle after chair restraint. Thus, apoSAA was predominantly associated with the denser HDL subfractions even though the lighter HDL subfractions were the most responsive in terms of changes in concentration. These data suggest that chair restraint of nonhuman primates induces apoSAA which displaces apoA-I and apoA-II or C's from HDL without altering the overall lipid and protein composition of the particle. In addition, chair restraint alters the concentration of HDL subfractions in ways that may be independent of apoSAA induction.     

Novel restraint system for neuroendocrine studies of socially living common marmoset monkeys

We describe a novel soft jacket and sling-harness restraint that permits species-typical postures for small-bodied primates, such as the common marmoset (Callithrix jacchus), during long-term (>6 h), continuous restraint. The restraint system is straightforward to use and manipulate, it is easily repaired, and the materials used are readily available. The soft jacket allows for increased versatility and longevity, and the sling-harness provides for greater movement and much longer duration of continuous restraint (up to 3 days) compared to a previously described, more conventional chair restraint for small-bodied primates. The new restraint system prevents the normal diurnal decrease in plasma cortisol levels across the daylight hours; however, it does not disrupt ovulatory cycles. Unlike the previously available techniques, therefore, this new restraint system is applicable to many neurobiological and neuroendocrine studies involving small-bodied, non-human primates and is especially suited to investigations requiring the maintenance of relationships within social groups.     

A restraint chair for primates

A humane, adjustable and comfortable restraint chair is described which is suitable for a wide variety of experiments on unsedated primates weighing between 3.5 and 15 kg.     

Use of a telemetry system to examine recovery of the cardiovascular system after excitement induced by handling stress in a conscious cynomolgus monkey (Macaca fascicularis)

The aim of this study was to determine the time required for the cardiovascular system of a conscious cynomolgus monkey, in which a telemetry had been implanted, to recover from excitement induced by handling stress. With enforcement of guidelines regarding safety pharmacological studies, cardiovascular studies in primates have become more important. However, as macaque monkeys are promptly excited under experimental procedures, it is often difficult to evaluate the drug effects on the cardiovascular system. Therefore, we tested monkey chair restraint and intravenous injection of saline. After monkey chair restraint and intravenous injection, approximately 30 minutes were required for recovery of both heart rate and blood pressure to their pre-treatment level; however, ECG parameters such as PR, QRS interval, and QTc did not drastically change. Based on our current results and with sufficient consideration of autonomic nervous effects, accurate evaluation of drug effects on the cardiovascular system should now be possible.     

The influence of physical and chemical restraint on the physiology of the chacma baboon (Papio ursinus)

The chacma baboon (Papio ursinus) is extensively used in South Africa for biomedical research. Being a large primate, it is always necessary to apply some measure of chemical or physical restraint. The physiological effects of placing an animal in a restraint chair are compared with the effects of various chemical agents, such as ketamine, halothane, and ketamine/xylazine combination over 90 min. It was found that ketamine and thiopentone infusion were a satisfactory chemical restraint agent that gave a stable physiological state over 90 min.     

The fine structural morphology of adrenal cortices of normal and stressed squirrel monkeys

The fine structural morphology of the male squirrel monkey adrenal cortex has been examined. When compared to other laboratory animals, the squirrel monkey adrenal cortex secretes large amounts of cortisol and maintains extraordinarily high plasma cortisol levels for prolonged periods of time. The normal cortical cells have numerous mitochondria with either a tubulo-vesicular or lamellar internal membrane arrangement, a well-developed agranular endoplasmic reticulum which is arranged in juxtaposition to mitochondria and lipid droplets, several lysosomes, and numerous thin-walled blood vessels of large caliber, suggestive of a rich blood flow through the gland. These characteristics have heretofore been associated with hypersecretion. Their presence in the squirrel monkey cortex, known to have high secretory activity, lends credence to the correlation of hyperdevelopment of the agranular reticulum with increased rates of secretion of corticoids. During chair restraint, the plasma cortisol levels rise two to three fold. Adrenocortical cells thus stressed exhibit a depletion and disorientation of membranes both of the agranular endoplasmic reticulum and mitochondria and a loss of ribosomes, lysosomes and, to some degree, intracellular lipid. The animal appears to be responding maximally to the stress of chair restraint. These fine structural characteristics are interpreted as an example of an adrenal cortex in the process of becoming functionally exhausted, since these animals sometimes do not survive the stress of chair restraint.     

Positive reinforcement training affects hematologic and serum chemistry values in captive chimpanzees (Pan troglodytes)

Positive reinforcement training (PRT) techniques have received considerable attention for their stress reduction potential in the behavioral management of captive nonhuman primates. However, few published empirical studies have provided physiological data to support this position. To address this issue, PRT techniques were used to train chimpanzees (Pan troglodytes) to voluntarily present a leg for an intramuscular (IM) injection of anesthetic. Hematology and serum chemistry profiles were collected from healthy chimpanzees (n=128) of both sexes and various ages during their routine annual physical examinations over a 7-year period. Specific variables potentially indicative of acute stress (i.e., total white blood cell (WBC) counts, absolute segmented neutrophils (SEG), glucose (GLU) levels, and hematocrit (HCT) levels) were analyzed to determine whether the method used to administer the anesthetic (voluntary present for injection vs. involuntary injection) affected the physiological parameters. Subjects that voluntarily presented for an anesthetic injection had significantly lower mean total WBC counts, SEG, and GLU levels than subjects that were involuntarily anesthetized by more traditional means. Within-subjects analyses revealed the same pattern of results. This is one of the first data sets to objectively demonstrate that PRT for voluntary presentation of IM injections of anesthetic can significantly affect some of the physiological measures correlated with stress responses to chemical restraint in captive chimpanzees.     

Acute restraint device for rhesus monkeys

A practical and inexpensive alternative to the standard primate chair is described. The apparatus is designed to allow easy removal of rhesus monkeys from their home cages and to allow restraint without anesthesia. A portable Plexiglas cage, which can be adjusted to accommodate rhesus monkeys of varying sizes, is placed against an animal's home cage. The animal is then trained to avoid the squeeze mechanism of the home cage by going into the Plexiglas cage. The animal's head can then be secured by means of poles hooked to a light weight collar worn permanently. This device and procedure allows an investigator to work with a restrained animal without resorting to drugs, unnecessary force or chronic restraint. Animals can be transferred daily with this technique with minimal conditioning and cooperation.     

Effect of Roman chair exercise training on the development of lumbar extension strength

The purpose of this study was to determine the effect of 45 degrees Roman chair exercise training on the development of lumbar extension strength. Fifteen healthy volunteers (9 women, 6 men) were recruited from a university setting and were randomly assigned to 1 of 2 groups. One group (n = 9) performed progressive resistance back extension exercise on a 45 degrees Roman chair once weekly for 12 weeks. Training consisted of one set of 8-20 dynamic repetitions to volitional exhaustion using hand-held metal plates for additional resistance. The other group did not train (control, n = 6). Peak isometric lumbar extension torque was measured on a lumbar extension dynamometer before and after the 12-week program. Following training, peak isometric lumbar extension torque did not increase for the Roman chair group (before: 224.0 +/- 134.1 N x m; after: 240.3 +/- 137.4 N x m; p > 0.05) compared with the control group (before: 175.6 +/- 68.9 N x m; after: 178.2 +/- 69.9 N x m; p > 0.05), despite an increase in dynamic exercise load.     

Using training to moderate chimpanzee aggression during feeding

The effects of human and nonhuman primate interactions on the well-being of captive nonhuman primates have been studied less rigorously than other forms of environmental improvement. Human intervention might be used to moderate severe aggression of socially housed animals at feeding times, one of the most common behavioral management problems. In this study, positive reinforcement training techniques were applied to reduce a dominant male chimpanzee's aggression and chasing during meals. Verbal commands and food reinforcers were used to train him to sit and remain seated while other group members received and ate their share of produce. Observational data were collected recording all instances of agonism in the group during periods when meals were fed and during times when no meals were offered, both before and after the training was accomplished. Multivariate analysis of variance for repeated measures results indicated that the training program successfully reduced the group's incidence of displaying, submission, and aggression during feeding periods. This effect was specific to the times meals were fed; there was no generalized reduction in the group's agonism after the training. This study exemplifies the use of positive reinforcement techniques to modify chimpanzee social dynamics during a problematic situation and of observational data to evaluate quantitatively the behavioral effects of the training. Training can be employed to help solve behavioral management problems among socially housed animals, and it can help improve the psychological well-being of captive chimpanzees. © 1994 Wiley-Liss, Inc.     

Training large macaws for artificial insemination procedures

For some endangered parrot species, captive breeding may be the only insurance for their survival. However, many individuals in captivity do not reproduce. Artificial insemination (AI) may help overcome reproductive failures or geographic distance. For semen collection in birds, massage is the most commonly used method. However, this process, which usually requires capture and restraint, involves risk of stress and injuries. The aim of this experiment was to train large macaws to accept the physical manipulations of their body parts needed for the artificial insemination process. Within 15 weeks, a male and a female Buffon's macaw (Ara ambiguus) learned to accept handling without apparent stress. A pair of green-winged macaws (Ara chloropterus) progressed more slowly and displayed some signs of stress. This stress highlights the need to monitor the birds' possible signs of discomfort during the training in order to adapt the working protocol. These results demonstrate that it is possible to train the AI behaviors, thus avoiding the capture, restraint, and anesthesia. An added benefit to this is the potential for the training to provide a form of behavioral enrichment.     

Human muscle power output during upper- and lower-body exercises

The purpose of this study was to evaluate the use of traditional resistance training equipment in the measurement of muscular power. This was accomplished by measuring the velocity of movement through a measured distance during maximal effort lifts using a Smith rack. The reliability of the method was established using 10 male volunteers who performed both bench press and squat exercises in a Smith rack. Maximal power output was determined at 30, 40, 50, 60, 70, 80, and 90% of the subject's 1 repetition maximum (1RM). Test-retest power values were not statistically different. Another 15 male volunteers who had previous muscle biopsy data from the vastus lateralis muscle performed the same maximal power output evaluation. There were no significant relationships between peak power outputs and fiber-type expressions when linear regressions were performed. The power curve produced by graphing power output vs. the percentage of 1RM indicates that peak power output occurs between 50 and 70% of 1RM for the squat and between 40 and 60% of 1RM for the bench press. These data indicate that this method of evaluation of muscle power is reliable, although it is not predictive of muscle fiber-type percentages.     

In vivo contact kinematics and contact forces of the knee after total knee arthroplasty during dynamic weight-bearing activities

Analysis of polyethylene component wear and implant loosening in total knee arthroplasty (TKA) requires precise knowledge of in vivo articular motion and loading conditions. This study presents a simultaneous in vivo measurement of tibiofemoral articular contact forces and contact kinematics in three TKA patients. These measurements were accomplished via a dual fluoroscopic imaging system and instrumented tibial implants, during dynamic single leg lunge and chair rising-sitting. The measured forces and contact locations were also used to determine mediolateral distribution of axial contact forces. Contact kinematics data showed a medial pivot during flexion of the knee, for all patients in the study. Average axial forces were higher for lunge compared to chair rising-sitting (224% vs. 187% body weight). In this study, we measured peak anteroposterior and mediolateral forces averaging 13.3% BW during lunge and 18.5% BW during chair rising-sitting. Mediolateral distributions of axial contact force were both patient and activity specific. All patients showed equitable medial-lateral loading during lunge but greater loads at the lateral compartment during chair rising-sitting. The results of this study may enable more accurate reproduction of in vivo loads and articular motion patterns in wear simulators and finite element models. This in turn may help advance our understanding of factors limiting longevity of TKA implants, such as aseptic loosening and polyethylene component wear, and enable improved TKA designs.     

The use of nonmetal electrodes in electroejaculation of restrained but unanesthetized macaques

A successful technique for electroejaculation with nonmetallic electrodes cut from defibrillation pads is described. Twenty-six adult male cynomolgus and eleven adult male rhesus macaques were electroejaculated while immobilized with chair restraint. From 123 attempted electroejaculations in both species of macaques, 119 semen specimens were obtained. The volume, concentration, % motility, and % normal forms of cynomolgus and rhesus macaque semen are presented. The use of nonmetallic electrodes provides a high quality ejaculate while minimizing the risks of adverse affects on valuable populations of macaques.     

Negative versus positive reinforcement: An evaluation of training strategies for rehabilitated horses

Rescued equids are often exposed to rehabilitation and training (or retraining) programmes to improve their physical and psychological well-being as well as to facilitate the re-homing process. Training uses either positive or negative reinforcement learning procedures and it is considered here that, there may be welfare implications associated with using the latter technique as it has the potential to overlay acute stress on animals with a chronic stress life history. The aim of this study, therefore, was to compare these training strategies (negative versus positive reinforcement) on equine behaviour and physiology as the first step in establishing an optimal rehabilitation approach (from a welfare perspective) for equids that have been subjected to chronic stress in the form of long-term neglect/cruelty. Over a 7-week period, 16 ponies (aged 6–18 months) were trained using either positive (‘positive’) (n = 8) or negative reinforcement (‘negative’) (n = 8) techniques to lead in hand, stand to be groomed, traverse an obstacle course and load into a trailer. Heart rate was measured (5 s intervals) on days 1 and 4 of each training week, ‘Pre’- (1 h), ‘During’ (0.5 h) and ‘Post’- (1 h) training session. Ethograms (10.00–20.00 h) outside of the training period were also compiled twice weekly. In addition, weekly arena tests (as a measure of reactivity) were also performed 1 week before and during the 7 weeks of training.Results showed significant differences between the two training schedules for some measures during the latter stages of the trial and suggested that animals trained under a positive reinforcement schedule were more motivated to participate in the training sessions and exhibited more exploratory or ‘trial and error’ type behaviours in novel situations/environments. In this context, the incorporation of positive reinforcement schedules within a rehabilitation programme may be of benefit to the animal from a welfare perspective.