Effects of post-treatment retention interval and context on neophobia and conditioned taste aversion

We have repeatedly observed that a delay between acquisition and test, and the nature of the context in which the delay is spent, modulates latent inhibition (LI) of conditioned taste aversion (CTA; e.g. [Anim. Learn. Behav. 28 (2000) 389; Anim. Learn. Behav. 30 (2002) 112]). The present paper analysed the effects of delayed testing and treatment context after flavor exposure on the recovery of neophobia (Experiment 1) and on extinction after simple conditioning (Experiment 2). Two experiments were conducted with the same factorial design (2x2: 1 day versus 21 days of delay between first and second stage, and home versus experimental cages as place of experimental treatment). There were independent effects of both variables on habituation of neophobia and conditioning strength as measured on extinction trials. The long delay produced a reduction of neophobia (Experiment 1) and an increase in conditioning (Experiment 2). In addition, more of the flavored solution was consumed when the experimental treatment was conducted in the home cage than in the experimental cage (Experiment 1), and there was stronger conditioning when the delay period took place in the experimental cages than in the home cages (Experiment 2). The implications of these results for LI, as well as their relevance for experiments that use the CTA paradigm, are discussed.     

Alterations in plasma catecholamines and behavior during acute stress in spontaneously hypertensive and Wistar-Kyoto normotensive rats

The responsiveness of the sympathoadrenal system to stress was assessed in spontaneously hypertensive (SHR) and Wistar-Kyoto normotensive (WKY) rats at 6, 18, and 48 weeks of age. Two days after insertion of a tail arterial catheter, each rat was transferred from its home cage to a shock chamber, and after 5 min received 60 footshocks over a 5 min interval. Blood samples were taken from undisturbed rats when in the home cage, 3–5 min after transfer to the shock chamber, and at the end of shock. An additional group of naive SHR and WKY rats was exposed to footshock and behavioral responses were recorded. There were no strain differences in levels of norepinephrine (NE) or epinephrine (EPI) while rats were undisturbed in their home cages. Transfer to the shock chamber resulted in a greater increase in plasma levels of both catecholamines in SHRs of each age. A similar pattern was evident after footshock; SHR rats had significantly higher post-shock levels of plasma NE and EPI than age-matched WKY rats. During shock, SHR rats were more active and jumped and reared more frequently than WKYs. These results demonstrate that the sympathoadrenal system of SHR rats is more responsive than normotensive rats to stressful stimuli and that this hyper-responsitivity is independent of increases in blood pressure. The excessive discharge of NE and EPI into plasma during stress may contribute to the development and maintenance of high blood pressure in SHR rats.     

The role of contextual cues in operant responding in rats

Following lever-press training on a variable-interval 60-second schedule of food presentation, groups of rats either remained in their home cages or were exposed to the operant chamber, from which lever and food had been removed, for five sessions. The lever was replaced in the chamber and rats from Group 1 (exposure to chamber) and Group 3 (home cage) were returned to the variable-interval schedule. Although response rates in test sessions were somewhat lower than at the end of training, there was no statistically significant difference in rates for either group. Rats in Group 2 (exposure to chamber) and Group 4 (home cage) received two test sessions of extinction. During the first session, Group 2 rates of lever pressing were significantly higher than Group 4 rates. These findings do not support the view that associations between contextual cues and the reinforcer serve to energize instrumental behavior (Pearce & Hall, 1979), and provide only minimal support for the view that contextual cues control responses that compete with the operant (Mills, 1980).     

Activity of anterolateral motor cortex neurons in instumental food-acquisition and alcohol-acquisition behavior

Singleunit activity of anterolateral area of motor cortex in rabbits subjected to chronic ethanol treatment was recorded to study interconnections of neuronal mechanisms of newly formed instrumental alcohol-acquisition behavior (IAB) and previously formed food-acquisition behavior (IFB). Adult animals were trained to perform IFB in experimental cage equipped with two food boxes and two pedals situated in the corners of the cage. Food was presented automatically in a food box after the pressing of an appropriate pedal. Same rabbits after 9 mo. of chronic alcohol treatment were trained to perform IAB in the same experimental cage (gelatin capsules filled with 15% ethanol solution were placed into the food box instead of food). Activity of 121 units of anterolateral area of motor cortex was studied. Each unit discharges were analysed in IAB as well as in IFB. The data obtained testifies that neuronal sets subserving IAB and IFB overlap but not completely. 44 "common" neurons permanently activated in both behaviors and 3 neurons specifically activated in each of behaviors (one in IAB and two in IFB) were found. We consider the formation of IAB as systemogenesis that is related to the consolidation processes: the formation of new neuronal specializations and to the accommodative re-consolidation: modification of early specialized cells ("common"). It is shown in the Discussion that present experiments help us not only understand interconnections of neuronal mechanisms of newly formed IAB and early formed IFB but also provide an additional insight into the nature of similarity between neuronal mechanisms of long-term memory and long-lived modifications resulting from repeated drug exposure.     

The experimental model of establishment and maintenance of social hierarchy in laboratory mice

The goal of the investigation is to establish a new experimental model of social hierarchy in laboratory mice. The important feature of the model is a group of two males only placed into the special experimental cage. The cage consists of two nestling chambers and central chamber. Observations were made 3 times per day (20 min each) throughout 5 days. Aggression, defence and sniffing behaviours of both partners were recorded during each observation period using video camera. Social rank was detected by asymmetry in aggressive behaviour. It was shown that social hierarchy was established during the first four tests limited by the first 24 h after pairing. After this period aggression and defence behaviours were greatly decreased. After the first 5 tests a stabilization in aggression (number of wins, active attacks and not direct aggression) and defence (number of vertical defensive postures and escapes) as well as sniffing behaviours took place. But a strong asymmetry in mentioned here behaviours in males of different ranks (dominants and subordinates) was kept up. Thus we offer a new experimental model of minimal society, which entirely reproduces a pattern of establishment and maintenance of social hierarchy. Our model demonstrates significant differences in agonistic, defence and sniffing behaviours between males of different social rank.     

Close Proximity of the Heterosexual Partner Reduces the Physiological and Behavioral Consequences of Novel-Cage Housing in Black Tufted-Ear Marmosets (Callithrix kuhli)

The present studies assessed the extent to which heterosexual pairmates could buffer marmosets (Wied's black tufted-ear marmoset,Callithrix kuhli)against stress. Six male and six female marmosets from established groups were exposed to two experimental manipulations together with a control condition. Each condition lasted a total of 4 days. For the two experimental conditions, animals were removed from the family group and housed in a novel cage for 48 h in either the presence or the absence of the heterosexual pairmate. During the 48-h novel-cage housing period and for 48 h upon reunion of the subjects with the family group, concentrations of urinary cortisol were measured in the first void sample of the day and behavioral observations were conducted. When animals were housed alone in a novel cage they exhibited significant elevations in levels of urinary cortisol after 24 and 48 h of novel-cage exposure. In contrast, when marmosets were housed in the novel cage in the presence of the pairmate, levels of urinary cortisol did not change across the 4-day period. The presence of the social partner also reduced the behavioral manifestations of exposure to novelty. Upon reunion with the family group, animals that had been housed in the novel cage alone spent significantly more time in close proximity to the pairmate than animals that had been housed with the partner. A second experiment was conducted to determine the effect that separation from the pairmate, only (independent of any effects of novelty), had on levels of cortisol. Concentrations of urinary cortisol were measured in subjects housed in the familiar home cage, but in the absence of the pairmate, over a 48-h period and compared to concentrations of excreted cortisol immediately prior to separation. Separation from the pairmate did not elevate cortisol levels when the subject was housed in the home cage, suggesting that elevated cortisol levels in animals housed alone in the novel cage were in response to novelty exposure rather than to separation from the pairmate. Since the physical presence of the heterosexual partner reduced the physiological and behavioral effects of novel-cage housing, social attachments might function as homeostatic regulators of HPA function in marmosets.     

Stress-induced hyperthermia depends on both time of day and light condition

Rats placed in an environment other than their home cage increase their body temperature (Tb) by more than 1 degree C. This stress-induced hyperthermia is considered to be a fever, in the sense that the Tb rise seems to reflect an upward shift in the level of regulated Tb (set point). The circadian rhythm of Tb also reflects changes in set point. One might therefore expect to see differences in response to such stress during various phases of the light-dark (LD) cycle as Tb fluctuates between L and D. To test this, 3- to 6-month-old male Long-Evans rats were taken from their home cages (12:12 LD) and placed individually in a Plexiglas container for 30 min. Tb and activity were measured via telemetry. In the first experiment, rats were placed in the container during day (from 1 to 3 h after lights on) and night (from 1 to 3 h after lights off), with light on or off during the test. There was a significant Tb rise in response to placement in the container at all times except when the rats were tested during the night with light on in the container; in that condition there was no Tb rise. In the second experiment, the authors determined that 30 min of light in the home cage before the test did not affect Tb: If the light was on in the test situation, hyperthermia was inhibited, and if it was off, hyperthermia was as high as control levels. In the third experiment, to determine whether this effect was time dependent, the test was performed at 4-h intervals, with light on or off during the test. The strongest inhibiting effect of light was in early night. In the fourth experiment, the authors turned the lights on during early night while the rats were in their home cages. This reduced their Tb significantly by less than 0.3 degrees C. The authors conclude that both clock time and light condition during testing are factors affecting the Tb rise in response to stress.     

Response of adult New Zealand white rabbits to enrichment objects and paired housing.

Enhancing the psychological well-being of laboratory animals has received much attention recently. Although many studies have been undertaken to determine the effects of cage enrichment techniques on dogs and nonhuman primates, other than scant empirical observations, little has been done to measure these events objectively in lagomorphs. We studied adult female New Zealand White (NZW) rabbits to learn if, when given the opportunity, individual rabbits would use different enrichment objects placed in their cages, and to determine if rabbits preferred to be in proximity to one another, or apart. Three different objects were evaluated with eight rabbits individually housed in conventional cages. Each object introduced into individual rabbit cages stimulated substantial interaction, especially chewing behavior. Eight other rabbits were pair-housed in a modified caging system with a special access port between two separate cages. When given a choice, rabbits preferred to be in the same cage with other rabbits. In both studies, individual behaviors were monitored, as well as either the type of interaction and percentage of observations spent with each object or, in the housing study, percentage of observations involved with different types of activity, and relative location of the paired rabbits.     

Assessing burrowing, nest construction, and hoarding in mice

Deterioration in the ability to perform "Activities of daily living" (ADL) is an early sign of Alzheimer's disease (AD). Preclinical behavioural screening of possible treatments for AD currently largely focuses on cognitive testing, which frequently demands expensive equipment and lots of experimenter time. However, human episodic memory (the most severely affected aspect of memory in AD) is different to rodent memory, which seems to be largely non-episodic. Therefore the present ways of screening for new AD treatments for AD in rodents are intrinsically unlikely to succeed. A new approach to preclinical screening would be to characterise the ADL of mice. Fortuitously, several such assays have recently been developed at Oxford, and here the three most sensitive and well-characterised are presented. Burrowing was first developed in Oxford. It evolved from a need to develop a mouse hoarding paradigm. Most published rodent hoarding paradigms required a distant food source to be linked to the home cage by a connecting passage. This would involve modifying the home cage as well as making a mouse-proof connecting passage and food source. So it was considered whether it would be possible to put the food source inside the cage. It was found that if a container was placed on the floor it was emptied by the next morning., The food pellets were, however, simply deposited in a heap at the container entrance, rather than placed in a discrete place away from the container, as might be expected if the mice were truly hoarding them. Close inspection showed that the mice were performing digging ("burrowing") movements, not carrying the pellets in their mouths to a selected place as they would if truly hoarding them. Food pellets are not an essential substrate for burrowing; mice will empty tubes filled with sand, gravel, even soiled bedding from their own cage. Moreover, they will empty a full tube even if an empty one is placed next to it. Several nesting protocols exist in the literature. The present Oxford one simplifies the procedure and has a well-defined scoring system for nest quality. A hoarding paradigm was later developed in which the mice, rather than hoarding back to the real home cage, were adapted to living in the "home base" of a hoarding apparatus. This home base was connected to a tube made of wire mesh, the distal end of which contained the food source. This arrangement proved to yield good hoarding behaviour, as long as the mice were adapted to living in the "home base" during the day and only allowed to enter the hoarding tube at night.     

Relationship of early infant state measures to behavior over the first year of life in the tufted capuchin monkey (Cebus apella)

Data on activity states were collected from 29 group-housed capuchin monkey (Cebus apella) infants for 3 h each week from birth to 11 weeks of age. The amounts of time spent in sleeping/drowsy, alert–quiet, and alert–active states were measured in these subjects. Videotaped observations of these infants were recorded 3 times/week in the home cage over the first year of life and were scored for a number of social and exploratory behaviors. The extent to which early infant activity state scores predicted later behavior in the home cage was examined. Infant state measures correlated significantly with home cage behavior during months 2–6 in that infants that had been more active in early infancy spent more time alone, with other animals, and in exploration and play and less time with mothers than did quieter infants. Early state measures were less successful in predicting home cage scores beyond 8 months of age, whereas differences in behavior attributable to housing variables became more salient in the latter part of the first year. There was also a negative correlation between mother and infant activity in months 2 and 3, in that more sedentary mothers tended to have more active infants. Am. J. Primatol. 44:43–56, 1998. © 1998 Wiley-Liss, Inc. This article is a U.S. Government work, and, as such, is in the public domain in the United States of America.     

Refining cage change routines: comparison of cardiovascular responses to three different ways of cage change in rats

Cage change is one of the unavoidable routines in laboratory rodent care. However, cage change disrupts the rodents' olfactory environment and can evoke stress reactions. In this study, the short-term cardiovascular responses to three different cage change procedures were compared with telemetric monitoring. These procedures were: placing the rats into a new, clean cage (NEW), transferring the old cage lid into the clean cage (LID) and transferring an enrichment object into the clean cage (ENR) with the animals. Seven outbred rats (four Hsd:Sprague-Dawley and three HsdBrlHan:WIST) were instrumented with telemetric transmitters. The reactions were recorded during the 24 h following the cage change procedures. All cage change procedures (and also simple handling) caused elevated heart rate and mean arterial pressure levels for up to 5 h after the procedure, with the largest effect seen during the first hour. The reactions observed after cage change were significantly (P < 0.05) greater than those observed after simple handling. The reactions after NEW were significantly higher than the reactions after ENR or LID, though the results were dependent on the stock. In Wistar rats the LID procedure resulted in smaller reactions than ENR. In Sprague-Dawley rats, the differences between ENR and LID were not so clear, but the transfer of scent-marked material into the new cage decreased the reactions compared with the NEW procedure also in this stock. Based on these results, using the old cage lid on the new cage could reduce the disturbance of cage change in rats.     

The effect of environmental enrichment on the behaviour of caged rabbits (Oryctolagus cuniculus)

Behaviour and use of the cage area were studied in 96 rabbits (Oryctolagus cuniculus) kept in an enriched cage system - with access to shelter and raised height at the back of the cage - and in a conventional cage system to estimate the effects of the environmental enrichment on the rabbits' welfare. The rabbits' behaviour and placement in the cage were observed, using continuous video recording through 24 h and direct scan sampling during the daytime. In addition, an open-field test was carried out with each rabbit, and after every single test, the rabbits' timidity of being captured was recorded. Rabbits kept in the conventional cage system, especially the females, showed more restlessness, excessive grooming, bar-gnawing and timidity than rabbits kept in the enriched cage system. This indicates increased stress in the rabbits kept in the conventional cage system. All the rabbits performed most of the active behavioural elements in the daytime and were resting mostly at night that shows that the rabbits in both cage systems were adapted to the daily activity in the animal unit; the enrichment had no effect on the daytime activity.Only a few rabbits, particularly the females, used the box as a shelter or resting-place. On the other hand, they more often used the roof of the box as a look-out or resting-place. Furthermore, the rabbits' behaviour showed that they utilised the raised height in the enriched cage system.These results indicate that rabbits kept in an enriched cage system, particularly the females, had better welfare than rabbits kept in a conventional cage system because they had access to shelter and a better chance of interacting with the environment.     

Latency to first naloxone-induced jump as a measure of precipitated abstinence intensity in morphine-dependent mice and the interaction of set on this test

A method developed in this laboratory uses latency to time of first jump after injection of naloxone rather than the number of jumps in a specified period or the number of animals jumping as a measure of the degree of morphine physical dependence. For the test, mice are placed in a glass cylinder used as a test chamber, after being injected with the antagonist. During the development of this method it was observed that repeated exposures of dependent mice to both naloxone and the chamber yielded shorter latencies to first jump than did repeated exposures to naloxone alone in animals with the same degree of physical dependence. It appears that learning develops when naloxone injections are given repeatedly and followed by exposure to the test chamber and that this learned behavior is manifested by a reduced latency to first jump which may be confused with increased intensity of the opiate-withdrawal syndrome.     

The reinforcing property and the rewarding aftereffect of wheel running in rats: a combination of two paradigms

Wheel running reinforces the behavior that generates it and produces a preference for the context that follows it. The goal of the present study was to demonstrate both of these effects in the same animals. Twelve male Wistar rats were first exposed to a fixed-interval 30 s schedule of wheel-running reinforcement. The operant was lever-pressing and the reinforcer was the opportunity to run for 45 s. Following this phase, the method of place conditioning was used to test for a rewarding aftereffect following operant sessions. On alternating days, half the rats responded for wheel-running reinforcement while the other half remained in their home cage. Upon completion of the wheel-running reinforcement sessions, rats that ran and rats that remained in their home cages were placed into a chamber of a conditioned place preference (CPP) apparatus for 30 min. Each animal received six pairings of a distinctive context with wheel running and six pairings of a different context with their home cage. On the test day, animals were free to move between the chambers for 10 min. Results showed a conditioned place preference for the context associated with wheel running; however, time spent in the context associated with running was not related to wheel-running rate, lever-pressing rate, or post-reinforcement pause duration.     

The newborn rat's stress system readily habituates to repeated and prolonged maternal separation, while continuing to respond to stressors in context dependent fashion

Adrenal corticosterone secretion of newborn mice rapidly desensitizes to repeated maternal absence. The present study investigated the effects of novelty exposure, maternal care and genotype on this phenomenon.Maternal separation (MS) took place on postnatal days (pnd) 3-5. In Wistar rats, the degree of novelty in the MS-environment was varied by exposing pups to: (i) “home separation”: pups remained in the home cage; (ii) “novel separation”: pups were placed individually in a novel cage. Maternal care was recorded on pnd 1 to 4. To investigate the effect of genotype, we also examined Long Evans in the “home separation” condition. Basal and stress-induced ACTH and corticosterone levels were measured. Adrenal tyrosine hydroxylase (TH) and melanocortin receptor-2 (MCR-2) proteins served as markers for adrenal function.We show, in both rat strains, that the rise in plasma corticosterone induced by a single 8 h-MS on pnd 5 was abolished, when this separation procedure had also been performed on pnd 3 and 4. Habituation to maternal absence occurred irrespective of housing conditions. However, pups in the “home separation” condition received less maternal care upon reunion than those placed in the “novel separation”. These “home separation” pups appeared more responsive to a subsequent acute novelty-stressor, and their adrenal TH and MCR-2 were higher. Long Evans rats appeared more stress responsive than the Wistars, in the home separation condition.In conclusion, separation environment, maternal care and genotype do not affect adrenal desensitization to repeated 8 h-MS itself, but may modulate the adrenal stress-responsiveness of separated pups.     

Ranging Patterns of a Western Gorilla Group During Habituation to Humans in the Dzanga-Ndoki National Park,Central African Republic

The ranging patterns of western gorillas have been described in relation to their more frugivorous diet as compared to mountain gorillas. However, the lack of data from habituated groups prevents a full understanding of their ranging behavior and its implications for western gorilla social behavior. Unhabituated gorillas often flee from approaching researchers, possibly resulting in longer daily ranges and home range expansions and/or shifts. I report on the influence of habituation on the ranging patterns of a western gorilla group. I analyzed progress in habituation through changes in gorilla reactions, displayed upon detecting the observers. I measured daily path length and home range while following the group's trails and monitored diet via fecal analyses and direct observations. During the study period, habituation progressed and the gorillas increasingly ignored the arrival of observers. As a result, daily path lengths were longer during the early stages of habituation, when the gorillas were avoiding observers. There was no range expansion or shift, though the measure is more difficult to investigate, being subject to error due to variation in sample size. We recorded seasonal changes in the consumption of fruit by gorillas, but no direct effect on ranging patterns is detectable with the data available, which is likely a consequence of the combined effect of habituation and seasonality on group movements. The findings are not only important for understanding the extent of human influence while conducting research on wild gorillas but also suggest the need for caution when interpreting results from non-habituated gorillas.     

Hyperactive behaviour in the mouse model of mucopolysaccharidosis IIIB in the open field and home cage environments

Mucopolysaccharidosis IIIB (MPS IIIB) is a lysosomal storage disorder characterized by severe behavioural disturbances and progressive loss of cognitive and motor function. There is no effective treatment, but behavioural testing is a valuable tool to assess neurodegeneration and the effect of novel therapies in mouse models of disease. Several groups have evaluated behaviour in this model, but the data are inconsistent, often conflicting with patient natural history. We hypothesize that this discrepancy could be due to differences in open field habituation and home cage behaviour. Eight-month-old wild-type and MPS IIIB mice were tested in a 1-h open field test, performed 1.5 h after lights on, and a 24-h home cage behaviour test performed after 24 h of acclimatization. In the 1-h test, MPS IIIB mice were hyperactive, with increased rapid exploratory behaviour and reduced immobility time. No differences in anxiety were seen. Over the course of the test, differences became more pronounced with maximal effects at 1 h. The 24-hour home cage test was less reliable. There was evidence of increased hyperactivity in MPS IIIB mice, however, immobility was also increased, suggesting a level of inconsistency in this test. Performance of open field analysis within 1-2 h after lights on is probably critical to achieving maximal success as MPS IIIB mice have a peak in activity around this time. The open field test effectively identifies hyperactive behaviour in MPS IIIB mice and is a significant tool for evaluating effects of therapy on neurodegeneration.     

Microclimate in two types of rat cages

The microclimate in two types of rat cages (a Makrolon type IV with a solid floor and a stainless steel cage with a wire mesh floor (five rats per cage)) placed in the same macro-environment was compared. The temperature, relative humidity and ammonia concentration in the cages were measured twice a day for 8 days. The cages were cleaned every 4 days. The greatest difference between the cage types was in the ammonia build-up. In Makrolon cages the ammonia concentration never reached 5 ppm, whereas in steel cages it showed a constant increase and already exceeded the threshold limit for man (25 ppm for 8 h per day) on the third day after cleaning.     

Fear responses of individually-caged laying hens as a function of cage level and aisle

Chickens kept under intensive conditions are often housed in multi-deck battery cage systems. The effects on egg production of housing birds at different cage levels have been extensively studied, but little is known of the behavioural consequences of this. The present study examined the responses of individually-caged adult laying hens from each tier of a 3-tier battery system in a number of situations intended to elicit fear. Birds housed in the top tier showed (a) longer durations of tonic immobility, (b) greater avoidance of a novel rod placed at the front of the cage, and (c) lower levels of approach and greater inhibition when placed in a pen containing either a human being or one of two novel, inanimate objects, than did those from middle and bottom tiers. The latter birds behaved remarkably similarly. The consistently higher fear scores recorded by hens from the top tier are considered likely to reflect differences in general fearfulness as a function of cage level, and may be a consequence of long-term exposure to different degrees of environmental stimulation.