Textbook recommendations for gavaging rats vary between 1-5 ml for an adult rat. Rats weighing either 130 g or 250 g were gavaged with varying dosages of barium sulphate (BaSO4). After dosing, radiographs were taken at 0, 15 and 60 min. Animals showing a section of the small intestine totally filled with BaSO4 were scored as displaying spontaneous release. Other rats of the same sizes were gavaged with similar doses and subsequently tested in an open-field arena for behavioural abnormalities that might indicate stress or pain resulting from the procedure. Body temperature before and after treatment was recorded using microchip transponders. None of the 250 g rats in the 1 ml dosage group showed spontaneous release through the pyloric sphincter. In the 2 ml and 4 ml dosage groups, only one out of five animals showed spontaneous release. In the 6 ml dosage group, half of the animals showed spontaneous release. In the 8 ml and 10 ml dosage groups, five out of six and four out of five, respectively, showed spontaneous release. If doses were higher than 12 ml, no animal was able to keep all of the BaSO4 in its stomach. In the rats weighing 130 g, the 3 ml dosage group showed only one out of four rats with spontaneous release, whereas in the 5 ml and 7 ml dosage groups, all animals showed spontaneous release. After 15 min, all of the rats in both weight groups showed BaSO4 in the duodenum. Ambulation, rearing up onto the hind legs and defecation, as well as body temperature immediately after dosing, correlated very strongly with the dose (ml kg−1); increasing the dose resulted in reduced ambulation, rearing, defecation and body temperature. However, 10 min after performance of the open-field test, neither body temperature, serum corticosterone nor serum glucose showed any correlation with dose. This study indicates that high doses (ie doses up to 10 ml for a 250 g rat) might be safe to use; however, if an adverse impact on the rat is to be avoided, use of much lower doses should be considered—for example, doses that do not enforce opening of the pyloric sphincter in any rat. This would be less than 4 ml kg−1 in a 250 g rat.

Most birds have visual sensitivity to ultraviolet (UV) wavelengths, and this sensitivity appears to play a role in their colour vision. Artificial lighting is normally deficient in UV wavelengths. Hence, there may be welfare implications for captive birds kept under such lighting. We investigated whether the absence of UV wavelengths during rearing adversely affects Japanese quail (Coturnix coturnix japonica). We also investigated the short-term effect of switching from UV-containing to UV-deficient lighting, and vice versa. Stress was assessed by monitoring behaviour and plasma corticosterone levels. We did not detect any significant difference in these variables between birds reared either with or without UV. We conclude that rearing quail in an absence of UV does not appear to have a significant impact on their welfare, as measured using these indicators.

Although genetic selection for certain characteristics has compromised the well-being of domestic animals, selective breeding could and probably should be used to improve welfare. Three of the major behavioural problems facing the poultry industry are fear, feather pecking and social stress. However, these and many other behavioural traits respond readily to genetic selection. The present paper reports the results of selective breeding studies in which underlying fearfulness, sociality, feather pecking, adrenocortical responsiveness, and growth rate were manipulated in Japanese quail, Coturnix japonica, or in chickens, Gallus gallus domesticus. The apparent benefits of selection for appropriate levels of these characteristics are discussed, as are the ethical issues involved. The ultimate aims of the selection programmes and the studies described here are to improve the birds ‘ welfare and productivity by increasing their ability to interact successfully with their physical environment, with one another, and with human beings.

Housing conditions can alter both the physiology and behaviour of laboratory animals. Forced-air-ventilated micro-isolation systems increase the efficient use of space, decrease the incidence of disease among laboratory rodents, and provide better working conditions for animal care staff; however, such systems can increase breeding variability and mortality. We examined the possibility that stressors associated with automated housing conditions evoke subtle changes among immune, endocrine, and behavioural parameters in mice housed in a static versus a forced-air-ventilated micro-isolation system. In addition, we assessed the effects of housing in the forced-air-ventilated micro-isolation system both with and without the use of an automatic watering system. Housing in the forced-air-ventilated micro-isolation system, using the automatic watering system, suppressed delayed-type hypersensitivity (DTH) responses, a measure of cell mediated immune function, compared with the responses of mice housed in static cages. Hypothalamic–pituitary–adrenal axis function was also altered by housing in the forced-air-ventilated micro-isolation system with the use of the automatic watering system, such that mice in this housing system had lower resting corticosterone concentrations and increased reactivity to restraint. Despite these changes in corticosterone, housing condition did not alter activity level or exploratory, anxiety-like, or depressive-like behaviours. These results suggest that investigators should carefully consider housing conditions in studies of immune and endocrine function.

Conventional bleeding of small laboratory animals is often associated with stress and injuries that can cause haematomas, inflammation and ultimately the death of animals under investigation. Here, we used faecal glucocorticoid metabolites as an indicator of stress imposed on laboratory mice (Mus musculus domesticus) when bled in three different ways: puncture of the tail vein following mechanical immobilisation; puncture of the retro-orbital plexus following chemical immobilisation; and a minimally invasive technique using blood-sucking bugs (Reduviidae, Heteroptera) without any immobilisation. We hypothesised that blood-sucking bugs provoke a lower hormonal stress response than conventional bleeding techniques because laboratory animals are not handled and because the mechanical stimulus of an insect sting is supposedly weak. Each of the 16 mice was bled using one of the three methods at a time in a random order with seven days of recovery between subsequent bleeding events. To monitor the stress hormones, we determined corticosterone metabolites in faecal samples of mice collected one day before, 8 h after and one day after the bleeding event. Concentrations of faecal glucocorticoid metabolites increased in all three treatment groups compared with baseline values. However, average concentration of stress hormone metabolites after bleeding was higher by a factor of about 1.5 when conventional bleeding techniques were applied than when bugs were used. We conclude that blood-sucking bugs may offer a gentle alternative for obtaining blood samples from small animals such as mice.

Amphibians, like other vertebrates, respond to acute stressors by releasing glucocorticoid steroid hormones that mediate physiological and behavioural responses to stress. Measurement of stress hormones provides a potential means to improve the welfare of laboratory animals. For example, manipulations of laboratory housing and procedures combined with measurement of glucocorticoids may identify which conditions are more stressful to animals. This is important because there is very little experimental evidence to guide best practice for welfare in amphibians and other lower vertebrates. We investigated the effect of different methods of euthanasia on the circulating plasma corticosterone levels in the African clawed frog (Xenopus laevis), a model amphibian organism that is frequently used in laboratories. In particular, we investigated the effect of different concentrations and pH of the anaesthetic tricaine methanesulphonate (MS-222). Low concentration and unbuffered (low pH) solutions of MS-222 caused elevated corticosterone levels, but only after the effect of MS-222 treatment on blood fluid volume had been taken into account. The level of disturbance that animals experienced also affected corticosterone levels. Thus, our data suggest that to minimise stress to X. laevis, animals should be euthanised after minimal disturbance and in a 3 g L−1 MS-222 solution, buffered to pH 7. The potential for the improvement of amphibian welfare using corticosterone measures as a tool is discussed.

Owl cafes, where customers view and interact with owls, have become popular in Japan. There are multiple aspects of the environment which may be stressful to nocturnal owls, including lighting, tethering and frequent interactions with humans but, to date, welfare has not been investigated. This preliminary study examines the effects of owl cafes and customers on the physiological stress (faecal corticosterone levels [FCL]) and behaviour of the owls. Seven eagle owls (Bubo bubo) and two African scops owls (Ptilopsis leucotis), in two cafes, were studied over an eight-day period. Cafe A (n = 5) was closed for one day per week, whilst cafe B (n = 4) was open every day. In cafe A there was higher FCL in owls during open days than closed days suggesting that the conditions on open days increase stress in owls. Eight of nine owls showed evidence of some aversion or avoidance of humans, whilst no affiliative behaviours were observed. The number of visitor interactions was not associated with the level of physiological stress; and when comparing owls, there was a tendency for individual owls with a higher percentage of aversive responses to customers to also have higher FCL. Close human interactions of a negative nature, that result in aversion, may be a significant stressor, but further research is required.

A multitude of endocrine mechanisms are involved in coping with challenges. Glucocorticoids, secreted by the adrenal glands, are in the front line of the battle to overcome stressful situations. They are usually measured in plasma samples as parameters of adrenal activity and thus of disturbance. Unfortunately, collecting blood samples itself can disturb an animal. Thus, non-invasive methods for the determination of glucocorticoids or their metabolites have become increasingly popular. The pros and cons of various non-invasive sample materials (saliva, excreta, milk, hair/feathers and eggs) for glucocorticoid determination are given. Above all, faecal samples offer the advantage that they can be collected easily. In faecal samples, circulating hormone levels are integrated over a certain period of time and represent the cumulative secretion of hormones. Thus, the levels are less affected by short fluctuations or the pulse-like nature of hormone secretion. However, using this technique to assess an animal's adrenocortical activity is not especially simple. Whether frequent sampling is necessary or single samples will suffice depends upon the study's aim (whether one is examining the impact of acute or chronic stressors). Background knowledge of the metabolism and excretion of cortisol/corticosterone metabolites is required and a careful validation for each species and sex investigated is obligatory. The present review also addresses analytical issues regarding sample storage, extraction procedures and immunoassays and includes a comprehensive list of published studies (up to 2011) describing the use of such methods in farmed animals. Applied properly, non-invasive techniques to monitor glucocorticoid metabolites in faecal samples of various species are a useful tool for welfare assessment, especially as they are easily applied at farm or group level.

Environmental adversity increases child susceptibility to disrupted developmental outcomes, but the mechanisms by which adversity can shape development remain unclear. A translational cross-species approach was used to examine stress-mediated pathways by which poverty-related adversity can influence infant social development. Findings from a longitudinal sample of low-income mother–infant dyads indicated that infant cortisol (CORT) on its own did not mediate relations between early-life scarcity-adversity exposure and later infant behavior in a mother-child interaction task. However, maternal CORT through infant CORT served as a mediating pathway, even when controlling for parenting behavior. Findings using a rodent “scarcity-adversity” model indicated that pharmacologically blocking pup corticosterone (CORT, rodent equivalent to cortisol) in the presence of a stressed mother causally prevented social transmission of scarcity-adversity effects on pup social behavior. Furthermore, pharmacologically increasing pup CORT without the mother present was not sufficient to disrupt pup social behavior. Integration of our cross-species results suggests that elevated infant CORT may be necessary, but without elevated caregiver CORT, may not be sufficient in mediating the effects of environmental adversity on development. These findings underscore the importance of considering infant stress physiology in relation to the broader social context, including caregiver stress physiology, in research and interventional efforts.

We previously reported that dietary vitamin E deficiency increased anxiety-like behaviour in rats exposed to social isolation. Here, we performed a detailed investigation of this phenomenon and its underlying mechanism. First, we fed Wistar rats with a vitamin E-free diet for 3 d, 1 week or 2 weeks and found an increase in anxiety-like behaviour after 1 and 2 weeks of vitamin E deficiency based on behavioural indicators. Next, we examined the effect of a control diet (150 mg all-racemic α-tocopheryl acetate/kg) on anxiety-like behaviours in rats that received a 4-week vitamin E-free diet. We found that increased anxiety-like behaviour was reversed to control levels after refeeding vitamin E for 7 d but not for 1 or 3 d. Further, anxiety-like behaviour increased or decreased gradually based on the amount of vitamin E intake; however, it had a quicker progression than physical symptoms of vitamin E deficiency. Moreover, rats fed with excess vitamin E (500 mg all-racemic α-tocopherol/kg diet) showed less anxiety-like behaviour than control rats, indicating that vitamin E supplementation is effective for preventing anxiety increase under social isolation stress. Since plasma corticosterone levels were higher in vitamin E-deficient rats, we investigated the effect of adrenalectomy on anxiety-like behaviour and found that adrenal hormones played an essential role in the increased anxiety-like behaviour induced by vitamin E deficiency. In conclusion, increased anxiety-like behaviour is a symptom that emerges earlier than physical vitamin E deficiency and is caused by adrenal hormone-dependent mechanisms.

Chronically stressed adult male Balb C mice were submitted to the tail suspension test. Chronic immobilization stress (6 h/d for 14 consecutive days) induced a significant reduction in immobility time when compared to non-stressed controls. Pretreatment with LY 53857, a serotonin 5HT2 antagonist, and IPS 339, a selective beta-2 adrenoceptor blocker, reversed immobility time to the levels of non-stressed controls. Chronic administration of corticosterone (100 mg/kg for 7 d) did not modify immobility time as compared to saline treated controls. It is suggested that both serotonergic and adrenergic pathways in the brain may participate in the stress-induced changes occurring in the tail suspension test response and that corticosterone does not appear to play a role in this process.

The novel antidepressant drug tianeptine had an antidepressant-like effect on a rat model of depression based on the deficit in open field activity observed on the day after 2 h immobilization. Thus, tianeptine (10 mg/kg ip) given 2 h after the end of the immobilization period led to the deficit being opposed. Similar results were obtained using rats previously given 10 mg/kg of the drug per day for 13 days and a final dose 2 h after the end of immobilization. Other workers have found that tianeptine attenuates stress-induced activation of the hypothalamo-pituitary-adrenal axis. Possible relationships between these findings are discussed together with the paradoxical co-existence of the antidepressant properties of tianeptine with its ability to decrease the availability of 5-HT to receptors.

The effect of the antidepressant agent, tianeptine, on the hypothalamo-pituitary-adrenal (HPA) axis was studied in adult male rats under basal and stressed conditions. Chronic treatment with tianeptine (10 mg/kg; 2 weeks; twice a day) induced a significant decrease of hypothalamic CRF (–12%) and pituitary ACTH concentrations (–21%), suggesting that tianeptine reduces the activity of hypothalamic CRF neurons and pituitary corticotrophs. The possible effect of tianeptine in the neuroendocrine response to stress was thus investigated. Rats were submitted to restraint stress for 30 min; under these conditions ACTH and corticosterone levels were considerably increased. A single injection of tianeptine was found to significantly reduce stress-evoked elevations of plasma ACTH and corticosterone. Time-course experiments, consisting of administering tianeptine 1–3 h before immobilization stress, revealed that the maximum inhibitory effect of tianeptine occurs about 2 h after injection of the antidepressant. Administration of various doses of tianeptine (from 2.5 to 20 mg/kg) showed that the effect of the drug was dose-dependent; the maximum effective dose being 10 mg/kg. Taken together, these data indicate that tianeptine may exert original ‘anti-stress’ activity.

Force-feeding was considered as a traditional high-efficiency approach to improve growth performance and accelerate fat deposition of Pekin ducks. However, force-feeding is a serious violation of international advocacy on animal welfare, because it can induce serious injuries to animals, such as damages to the digestive tract, effects on immunity and even severe oxidative stress. Therefore, it is urgent to stop force-feeding. The aim of this study was to determine the effects of force feeding on immune function, digestive function and oxidative stress in the mucosa of duodenum and jejunum of Pekin ducks. A total of 500 ducks were randomly divided into two groups. The control group was allowed to feed freely on a basal diet. The experimental group was force-fed by inserting a plastic feeding tube 8 to 10 inches long down the esophagus for 6 days. Compared with the control group, there was a significant (P<0.05) increase in serum diamine oxidase, d-lactic acid, endotoxin and corticosterone levels in the force-feeding group. The crypt depth in duodenum and jejunum showed significant differences (P<0.05) between the two groups and the intestinal villus epithelium cell was severely damaged in force-feeding group. Similarly, the activities of digestive enzymes as well as the levels of immune function in the duodenal and jejunal mucosa in the force-feeding group were significantly higher than the control group (P<0.05). However, there was a significant decrease in the superoxide dismutase, glutathione peroxidase and catalase levels with a marked increase in malondialdehyde level in duodenal and jejunal mucosa (P<0.05). In summary, at the end of the fattening period with force-feeding for 6 days, Pekin ducks experienced an adverse effect on the integrity of their duodenal and jejunal mucosa epithelium cell as well as their immune function and antioxidant capacity of Pekin ducks but also had improvement in digestive enzyme activities.

Free-range laying hen systems are increasing within Australia. The pullets for these systems are typically reared indoors before being provided first range access around 21 to 26 weeks of age. Thus, the rearing and laying environments are disparate and hens may not adapt well to free-range housing. In this study, we reared 290 Hy-Line® Brown day-old chicks divided into two rooms each with feed, water and litter. In the enriched room, multiple structural, manipulable, visual and auditory stimuli were also provided from 4 to 21 days, the non-enriched room had no additional objects or stimuli. Pullets were transferred to the laying facility at 12 weeks of age and divided into six pens (three enriched-reared, three non-enriched-reared) with identical indoor resources and outdoor range area. All birds were first provided range access at 21 weeks of age. Video observations of natural disturbance behaviours on the range at 22 to 23 and 33 to 34 weeks of age showed no differences in frequency of disturbance occurrences between treatment groups (P=0.09) but a decrease in disturbance occurrences over time (P<0.0001). Radio-frequency identification tracking of individually tagged birds from 21 to 37 weeks of age showed enriched birds on average, spent less time on the range each day (P<0.04) but with a higher number of range visits than non-enriched birds from 21 to 24 weeks of age (P=0.01). Enriched birds accessed the range on more days (P=0.03) but over time, most birds in both treatment groups accessed the range daily. Basic external health scoring showed minimal differences between treatment groups with most birds in visibly good condition. At 38 weeks of age all birds were locked inside for 2 days and from 40 to 42 weeks of age the outdoor range was reduced to 20% of its original size to simulate stressful events. The eggs from non-enriched birds had higher corticosterone concentrations following lock-in and 2 weeks following range reduction compared with the concentrations within eggs from enriched birds (P<0.0001). Correspondingly, the enriched hens showing a greater increase in the number of visits following range area reduction compared to non-enriched hens (P=0.02). Only one rearing room per treatment was used but these preliminary data indicate 3 weeks of early enrichment had some long-term effects on hen ranging behaviour and enhanced hen’s adaptability to environmental stressors.

Amphibians are considered to be the most threatened group of vertebrates. Among the multiple factors involved in their decline, habitat loss and alteration as a result of human activities is a major threat. At the individual level the effects of habitat alteration are potentially multiple, including a range of morphological and physiological responses. Analysing and understanding these responses is therefore a critical challenge for amphibian conservation. We examined the influence of intensive vehicle traffic (motorbikes and trucks on unpaved pathways) on the body size and condition and on the production of glucocorticoids (i.e. corticosterone) in the yellow-bellied toad Bombina variegata. In particular, we tested the hypothesis that intensive vehicle traffic has a negative influence on body size and body condition, and postulated that it also increases corticosterone production. Using morphometric data and saliva samples collected from four populations in France, we found that intensive vehicle traffic is associated with a decrease in body size and body condition in both males and females. Furthermore, our analysis revealed that corticosterone production was lower in both sexes in populations experiencing intensive vehicle traffic. We suggest that measures should be applied to reduce vehicle traffic intensity on unpaved pathways during toad breeding activity. This is critical for B. variegata, for which man-made ruts and residual puddles could mitigate the loss of natural habitats.

The probability that an individual will suffer from a major depressive episode is often considered to be influenced by risk factors such as gender, premature parental loss, exposure to pathogenic parental rearing, personality, a history of traumatic events, a previous history of major depression, low social support, recent stressful life events and difficulties and predisposing genetic influences. Although several studies suffer from methodological limitations, major depression (endogenous depression) is regarded as a multifactorial disorder and understanding its etiology requires the rigorous integration of several risk factors.