Non-enriched Animals Research Articles

Differential impact of shorter and longer periods of environmental enrichment on adult zebrafish exploratory activity (Danio rerio) in the novel tank paradigm

Several studies have used zebrafish to investigate the effects of environmental enrichment on behavior and physiology. However, to date there are no studies evaluating the behavioral responses, such as habituation and exploration, of enriched-housed zebrafish when they are submitted to novelty paradigms. The present work was, therefore, designed to evaluate the habituation and exploratory responses of zebrafish exposed to enriched- (EE) and non-enriched (NE) environments when they face novelty. Adult wild-type zebrafish were used. Three different enriched contexts were designed. In Context 1, zebrafish was exposed to enrichment during 7 days, which reduced their total distance traveled in novel tank and social preference tests in comparison to the non-enriched animals. In Context 2, animals were exposed to same enrichment during 14 days. EE exposure did not alter the behavioral responses of zebrafish compared to NE. In Context 3, fish were exposed to enrichment during 14 days, with changing the enriching elements at day 8. Similarly to Context 1, total distance traveled was reduced by EE exposure when compared to NE. Our results suggest a modulatory effect of EE on adult zebrafish locomotion that may be dependent on the time of exposure and on the physical structure of the enriched environment.

Positive Welfare and the Like: Distinct Views and a Proposed Framework.

Positive welfare and related terms such as good welfare, happiness, and a good life are increasingly used in the animal welfare science literature. Overall, they highlight the welfare benefits of providing animals opportunities for positive experiences, beyond the alleviation of suffering. However, the various terms remain loosely defined and are sometimes used interchangeably, resulting in discrepancy. In this perspective article, we lay out the terms and concepts used in the literature. We identify two distinct views: “hedonic positive welfare,” arising from likes and wants and their positive outcomes on welfare; and “positive welfare balance,” as an overall positive welfare state based on positive experiences outweighing negative ones. Eudaimonia, satisfaction with one's life, may emerge as a third view. We propose a framework that is applicable across the different views. The “Vienna Framework” outlines different facets: frequency, duration, arousal, context, previous experience, individual differences, sense of agency, and long-term benefit. The framework aims to encourage researchers to consider the relevance of these facets for their own research, to indicate how the facets are affected by different interventions (e.g., greater sense of agency in enriched compared to non-enriched animals), or to compare different topics with respect to the different facets (e.g., high arousal of play behavior and low arousal of social affiliation). We encourage researchers to carefully consider and clearly state how their work falls along these views and facets, conceptually, and operationally. This should prevent dilution of the meaning of positive welfare and thereby preserve its potential to improve the welfare of animals.

Housing condition-related changes involved in reversal learning and its c-Fos associated activity in the prefrontal cortex

Our study examined how different housing conditions modulated the acquisition of a spatial reference memory task and also, a reversal task in the 4-radial arm water maze (4-RAWM). The animals were randomly assigned to standard or enriched cages, and, as a type of complementary stimulation along with the environmental enrichment (EE), a group of rats also ran 15min/day in a Rotarod. Elevated-zero maze results allowed us to discard that our exercise training increased anxiety-related behaviors. 4-RAWM results revealed that the non-enriched group had a worse performance during the acquisition and also, during the first trial of each session with respect to the enriched groups. Regarding the reversal task, this group made more perseverative errors in the previous platform position. Interestingly, we hardly found differences between the two enriched groups (with and without exercise). We also analyzed how the reversal learning, depending on the previous housing condition, modulated the expression of c-Fos-positive nuclei in different subdivisions of the medial prefrontal cortex (cingulate (Cg), prelimbic (PL) and infralimbic (IL) cortices) and in the orbitofrontal (OF) cortex. The enriched groups had higher c-Fos expression in the Cg and OF cortices and lower in the IL cortex respect to the non-enriched animals. In the PL cortex, we did not find significant differences between the groups that performed the reversal task. Therefore, our short EE protocol improved the performance in a spatial memory and a reversal task, whereas the exercise training, combined with the EE, did not produce a greater benefit. This better performance seemed to be related with the specific pattern of c-Fos expression in brain regions involved in cognitive flexibility.

Increase of glucocorticoid receptor expression after environmental enrichment: Relations to spatial memory, exploration and anxiety-related behaviors

Environmental enrichment (EE) produces a remarkable degree of structural and functional plasticity in the hippocampus and possible mediators of these changes, such as glucocorticoid receptors (GRs), are of considerable interest. GRs are richly expressed in the hippocampus and they are involved in the adaptation to stressors and facilitate active coping in anxious situations. In this study, we assessed the effect of an EE protocol (24h/day during 69days) in adult Wistar rats on the activity in the elevated-zero maze (EZM), performance in the holeboard task (HB) and we also examined the changes in the glucocorticoid receptors (GRs) expression in the dorsal hippocampus (CA1, CA3 and DG). Our EE protocol reduced anxious behaviors in the EZM, so the animals spent more time and made more entries into the open sections. In the HB task, the enriched group showed more explorative behavior, a reduction of anxiety-related behaviors and a better cognitive performance compared to non-enriched animals. With regard to the GR expression, the EE condition produced an increase in the number of immunopositive cells for GRs in CA1, CA3 and DG. These results suggest that the better performance of enriched animals could be mediated in part by the increase of GRs in the dorsal hippocampus, which may alter the hippocampal neuronal function and accordingly, the anxiety levels, the spatial memory performance and the exploration levels in these animals.

Environmental enrichment extends ocular dominance plasticity into adulthood and protects from stroke-induced impairments of plasticity

Ocular dominance (OD) plasticity in mouse primary visual cortex (V1) declines during postnatal development and is absent beyond postnatal day 110 if mice are raised in standard cages (SCs). An enriched environment (EE) promotes OD plasticity in adult rats. Here, we explored cellular mechanisms of EE in mouse V1 and the therapeutic potential of EE to prevent impairments of plasticity after a cortical stroke. Using in vivo optical imaging, we observed that monocular deprivation in adult EE mice (i) caused a very strong OD plasticity previously only observed in 4-wk-old animals, (ii) restored already lost OD plasticity in adult SC-raised mice, and (iii) preserved OD plasticity after a stroke in the primary somatosensory cortex. Using patch-clamp electrophysiology in vitro, we also show that (iv) local inhibition was significantly reduced in V1 slices of adult EE mice and (v) the GABA/AMPA ratio was like that in 4-wk-old SC-raised animals. These observations were corroborated by in vivo analyses showing that diazepam treatment significantly reduced the OD shift of EE mice after monocular deprivation. Taken together, EE extended the sensitive phase for OD plasticity into late adulthood, rejuvenated V1 after 4 mo of SC-rearing, and protected adult mice from stroke-induced impairments of cortical plasticity. The EE effect was mediated most likely by preserving low juvenile levels of inhibition into adulthood, which potentially promoted adaptive changes in cortical circuits.

Enrichment and Training Improve Cognition in Rats with Cortical Malformations

Children with malformations of cortical development (MCD) frequently have associated cognitive impairments which reduce quality of life. We hypothesized that cognitive deficits associated with MCD can be improved with environmental manipulation or additional training. The E17 methylazoxymethanol acetate (MAM) exposure model bears many anatomical hallmarks seen in human MCDs as well as similar behavioral and cognitive deficits. We divided control and MAM exposed Sprague-Dawley rats into enriched and non-enriched groups and tested performance in the Morris water maze. Another group similarly divided underwent sociability testing and also underwent Magnetic Resonance Imaging (MRI) scans pre and post enrichment. A third group of control and MAM rats without enrichment were trained until they reached criterion on the place avoidance task. MAM rats had impaired performance on spatial tasks and enrichment improved performance of both control and MAM animals. Although MAM rats did not have a deficit in sociability they showed similar improvement with enrichment as controls. MRI revealed a whole brain volume decrease with MAM exposure, and an increase in both MAM and control enriched volumes in comparison to non-enriched animals. In the place avoidance task, MAM rats required approximately 3 times as long to reach criterion as control animals, but with additional training were able to reach control performance. Environmental manipulation and additional training can improve cognition in a rodent MCD model. We therefore suggest that patients with MCD may benefit from appropriate alterations in educational strategies, social interaction and environment. These factors should be considered in therapeutic strategies.

Environmental enrichment protects against functional deficits caused by traumatic brain injury

Environmental enrichment (EE) increases cortical weight, neuronal density, dendritic branching, and angiogenesis, all of which may be critical for functional recovery following insult. Our study was designed to determine possible benefits of pre-exposure to EE in preventing functional deficits following traumatic brain injury (TBI) to the prefrontal cortex. To examine the benefit of EE, adult male rats were placed in an enriched environment for 15 days. Enrichment was provided through social interaction, exercise, olfactory stimulation, and new objects/toys to explore. Following enrichment, experimental and age-matched controls were subjected to a moderate medial prefrontal cortex injury via controlled cortical impact (CCI). After 1 week recovery, animals were behaviorally tested to assess memory, anxiety, and sensory neglect. Lesion-induced deficits in spatial memory [Morris water maze (MWM)] were significantly attenuated in EE pre-exposed rats 18–21 days following injury. In addition, TBI-induced sensory neglect was significantly reduced in EE rats relative to non-enriched animals. No differences in anxiety-like behavior on the elevated plus maze (EPM) were detected. The behavioral data suggest that EE is neuroprotective when applied prior to TBI, resulting in improved recovery following injury.

Environmentally enriched rearing environments reduce repetitive perseveration in caged mink, but increase spontaneous alternation

Studies spanning 15 species (including American mink, Neovison vison) demonstrate that within similarly-housed populations, individuals displaying high levels of stereotypic behaviour (SB) typically show perseverative responding (e.g. during set-shifting, or reversal/extinction learning). Similar correlations in autism and schizophrenia suggest this indicates captivity-induced cortico-striatal circuit dysfunction. However, this pattern does not prove developmental impairment: SB, perseveration and their inter-correlations also occur in normal humans. We therefore differentially-reared enriched versus non-enriched mink to investigate whether treatments that exacerbate SB correspondingly increase perseveration (Study 1). Enriched-rearing did reduce SB and perseverative response repetition (in two-choice guessing tasks), while increasing spontaneous alternation: a strategy yielding more rewards, and suggesting enhanced hippocampal development. This complements previous research demonstrating cortical/hippocampal impairments and reduced behavioural flexibility in non-enriched animals, with implications for research animals and wild animals captive-raised for reintroduction into nature. Consistent with previous data, highly stereotypic subjects repeated guessing task responses most rapidly, suggesting disinhibition during repetition. However, unexpectedly, SB and perseveration did not co-vary across individuals. We therefore suggest that behavioural changes manifest as increased perseveration are important but do not fully explain captive animals’ SBs, possible reasons including the contributory role of differential motivations for underlying source behaviours. Re-analyses of old data (Study 2) confirmed that spontaneous alternation is profitable; and demonstrated that the precise methods used for quantifying perseveration and SB can modify the strength of apparent relationships between them, as can statistically controlling for feeding motivation: as predicted, partialling out motivational effects increased the variance in SB predicted by perseveration.

Both increases in immature dentate neuron number and decreases of immobility time in the forced swim test occurred in parallel after environmental enrichment of mice

A direct relation between the rate of adult hippocampal neurogenesis in mice and the immobility time in a forced swim test after living in an enriched environment has been suggested previously. In the present work, young adult mice living in an enriched environment for 2 months developed considerably more immature differentiating neurons (doublecortin-positive, DCX +) than control, non-enriched animals. Furthermore, we found that the more DCX + cells they possessed, the lower the immobility time they scored in the forced swim test. This DCX + subpopulation is composed of mostly differentiating dentate neurons independently of the birthdates of every individual cell. However, variations found in this subpopulation were not the result of a general effect on the survival of any newborn neuron in the granule cell layer, as 5-bromo-2-deoxyuridine (BrdU)-labeled cells born during a narrow time window included in the longer lifetime period of DCX + cells, were not significantly modified after enrichment. In contrast, the survival of the mature population of neurons in the granule cell layer of the dentate gyrus in enriched animals increased, although this did not influence their performance in the Porsolt test, nor did it influence the dentate gyrus volume or granule neuronal nuclei size. These results indicate that the population of immature, differentiating neurons in the adult hippocampus is one factor directly related to the protective effect of an enriched environment against a highly stressful event.

Environmental enrichment effects in social investigation in rats are gender dependent

Environmental enrichment (E) can change neuronal structure, improves learning in various tasks and increases cerebral plasticity. However, no E effects were found in a test measuring social olfactory discrimination memory and, although they have been mainly measured in males, gender differences have been reported in other tests. The aims of the present study were to evaluate gender differential effects of E in the social discrimination paradigm which also involves social olfactory discrimination and in the elevated plus-maze test (EPM) for measuring anxiety. E procedure consisted of a combination of social and physical factors; groups of 11–12 Sprague–Dawley rats were separated by sex in large cages with physical stimulus for a period of 8 weeks starting immediately after weaning. Differential gender E effects appeared in the social exploratory patterns: enriched males showed increased exploratory behaviour towards juvenile rats in comparison to control males, whereas no differences were found in females. No effects of E in social discrimination memory were observed. In the EPM, both enriched male and female rats showed less anxious behaviour than non-enriched animals.

Preweaning enrichment has no lasting effects on adult hippocampal neurogenesis in four‐month‐old mice

Since both living in an enriched environment and physical activity stimulate hippocampal neurogenesis in adult mice, we endeavored to examine whether pre-weaning enrichment, a sensory enrichment paradigm with very limited physical activity, had similar effects on neurogenesis later in life. Mice were removed from the dams for periods of increasing length from post-natal day 7 to 21, and exposed to a variety of sensory stimuli. At the age of 4 months, significant differences could be found between previously enriched and nonenriched animals when spontaneous activity was monitored. Enriched mice moved longer distances, and spent more time in a defined center zone of the open field. Adult neurogenesis was examined by labeling proliferating cells in the dentate gyrus with bromodeoxyuridine (BrdU). Cell proliferation, survival of the newborn cells, and net neurogenesis were similar in both groups. Volumetric measurements and stereological assessment of total granule cell counts revealed no difference in size of the dentate gyrus between both groups. Thus, in contrast to postweaning enrichment, preweaning enrichment had no lasting measurable effect on adult neurogenesis. One of the parameters responsible for this effect might be the lack of physical activity in preweaning enrichment. As physical activity is an integral part of postweaning enrichment, it might be a necessary factor to elicit a neurogenic response to environmental stimuli. The result could also imply that baseline adult hippocampal neurogenesis is independent of the changes induced by preweaning enrichment and might not contribute to the sustained types of plasticity seen in enriched animals.

The effects of environmental enrichment on voluntary ethanol consumption and stress ulcer formation in rats

The effects of exposure to an enriched environment on subsequent voluntary ethanol intake and response to restraint stress were examined. Rats at 21 days of age were reared in an enriched environment for 90 days. Non-enriched animals were reared individually in standard laboratory cages. Following an initial 36 day ethanol exposure period, voluntary ethanol (9% v/v) preference was assessed for 10 days. In addition, at the conclusion of the ethanol test session, animals were exposed to restraint stress for a 3 hr period. Results indicate that exposure to an enriched environment produces increased voluntary ethanol consumption as compared to non-enriched controls. Furthermore, animals reared in the enriched environment demonstrated reduced gastric ulcer severity in response to restraint stress. Ethanol per se did not affect ulcer formation.

Plasticity in the 904-day-old male rat cerebral cortex

Ten pairs of male Long-Evans rats living in nonenriched environments (3 rats per small cage) were transferred to either enriched environments (10 rats per large cage plus “toys”) or nonenriched environments (2 rats per small cage) at 766 days of age. One hundred and thirty-eight days later, at 904 days of age, the cerebral cortical thickness from these animals was measured on projected, 10-μm, thioninestained, transverse sections. Although the thickness in the enriched rats was greater than in the nonenriched rats in all sections through the frontal, parietal, and occipital cortices, the 4 to 10% differences were statistically significantly different in only the frontal and occipital cortices. Right greater than left cortical thickness differences were not statistically significant in either the enriched or the nonenriched animals by 904 days of age. Neuron and glial counts were made on enlarged photographs of area 18 in the occipital cortex on 6-μm-thick, luxol fast blue-stained sections. No significant differences in cell counts were noted between the enriched and nonenriched animals. No significant differences in neuronal counts were found among 108-, 650- (from previous experiments), and the 904-day-old nonenriched rats. The notable findings were the plasticity of the extremely old, enriched rats' occipital cortex and the lack of the loss of neurons in cerebral cortical area 18, whether or not the environments were enriched. These results showed that the cerebral cortex remained structurally plastic throughout the lifetime of the organism.

The role of motor learning and cage size in the early enrichment effect in mice.

Three experiments with C57BL/10J mice examined the possible roles of cage size and simple motor practice as factors responsible for producing improved performance of animals reared in enriched environments. Neither factor was found sufficient to improve subsequent performance in a food-seeking task. Mice reared in flat environments containing a variety of objects, but designed to prevent any climbing practice, out-performed nonenriched animals on a task requiring extensive climbing activity.