Optimal sampling interval for characterisation of the circadian rhythm of body temperature in homeothermic animals using periodogram and cosinor analysis.

Core body temperature (T c) is a critical aspect of homeostasis in birds and mammals and is increasingly used as a biomarker of the fitness of an animal to its environment. Periodogram and cosinor analysis can be used to estimate the characteristics of the circadian rhythm of T c from data obtained on loggers that have limited memory capacity and battery life. The sampling interval can be manipulated to maximise the recording period, but the impact of sampling interval on the output of periodogram or cosinor analysis is unknown. Some basic guidelines are available from signal analysis theory, but those guidelines have never been tested on T c data. We obtained data at 1-, 5- or 10-min intervals from nine avian or mammalian species, and re-sampled those data to simulate logging at up to 240-min intervals. The period of the rhythm was first analysed using the Lomb-Scargle periodogram, and the mesor, amplitude, acrophase and adjusted coefficient of determination (R 2) from the original and the re-sampled data were obtained using cosinor analysis. Sampling intervals longer than 60 min did not affect the average mesor, amplitude, acrophase or adjusted R 2, but did impact the estimation of the period of the rhythm. In most species, the period was not detectable when intervals longer than 120 min were used. In all individual profiles, a 30-min sampling interval modified the values of the mesor and amplitude by less than 0.1°C, and the adjusted R 2 by less than 0.1. At a 30-min interval, the acrophase was accurate to within 15 min for all species except mice. The adjusted R 2 increased as sampling frequency decreased. In most cases, a 30-min sampling interval provides a reliable estimate of the circadian T c rhythm using periodogram and cosinor analysis. Our findings will help biologists to select sampling intervals to fit their research goals.

Behavioural and physiological responses to stressors in sheep with temperament classified by genotype or phenotype.

The single nucleotide polymorphism (SNP) rs107856856, located in the tryptophan hydroxylase-2 gene, is associated with the behavioural phenotype for sheep temperament measured at weaning. Here, we tested the association between that SNP and physiological and behavioural responses to stressors in adult sheep. Two groups of adult sheep, one with genotype A/A (calm genotype) and the other with G/G (nervous genotype) in rs107856856, were selected from 160 sheep and were exposed, twice, to an open-field arena and an isolation box test (IBT). During each repeat, the behaviour and physiological responses (cortisol, prolactin, dehydroepiandrosterone [DHEA], brain derived neurotrophic factor [BDNF], characteristics of the response of body temperature, and oxidative stress) were measured. The behavioural and physiological responses of the sheep were compared between genotypes and also between groups classified on their phenotype as assessed by their initial isolation box score ("low responders" and "high responders"). The SNP rs107856856 had some effects on the behavioural phenotype (IBT score) but no effects on the physiological response to stress (cortisol, prolactin, DHEA, BDNF, oxidative stress or changes in body temperature) in the adult sheep, probably because the sheep were exposed, and therefore had adapted, to human contact during their life.

Functional interactions between coat structure and colour in the determination of solar heat load on arid living kangaroos in summer: balancing crypsis and thermoregulation.

Interactions of solar radiation with mammal fur are complex. Reflection of radiation in the visible spectrum provides colour that has various roles, including sexual display and crypsis, i.e., camouflage. Radiation that is absorbed by a fur coat is converted to heat, a proportion of which impacts on the skin. Not all absorption occurs at the coat surface, and some radiation penetrates the coat before being absorbed, particularly in lighter coats. In studies on this phenomenon in kangaroos, we found that two arid zone species with the thinnest coats had similar effective heat load, despite markedly different solar reflectances. These kangaroos were Red Kangaroos (Osphranter rufus) and Western Grey Kangaroos (Macropus fuliginosus).Here we examine the connections between heat flow patterns associated with solar radiation, and the physical structure of these coats. Also noted are the impacts of changing wind speed. The modulation of solar radiation and resultant heat flows in these coats were measured at wind speeds from 1 to 10 m s-1 by mounting them on a heat flux transducer/temperature-controlled plate apparatus in a wind tunnel. A lamp with a spectrum like solar radiation was used as a proxy for the sun. The integrated reflectance across the solar spectrum was higher in the red kangaroos (40 ± 2%) than in the grey kangaroos (28 ± 1%). Fur depth and insulation were not different between the two species, but differences occurred in fibre structure, notably in fibre length, fibre density and fibre shape. Patterns of heat flux within the species' coats occurred despite no overall difference in effective solar heat load. We consider that an overarching need for crypsis, particularly for the more open desert-adapted red kangaroo, has led to the complex adaptations that retard the penetrance of solar radiation into its more reflective fur.

Social media as a tool to understand the distribution and ecology of elusive mammals.

Comparatively little is known about the distribution and ecology of Aardvark (Orycteropus afer) and Temminck's Ground Pangolin (Smutsia temminckii). Both are elusive species that are normally nocturnal, solitary, and fossorial. Formally collected records have been used to map the distribution of these species, and social media records provide a tool to gather information on their distribution and ecology. We obtained 680 photographs and videos of aardvarks and 790 of ground pangolins in southern Africa from publicly available posts on Facebook and Instagram (2010-2019). The images provide new insights into the distribution, activity, drinking, and predation-and confirm that aardvarks are more diurnally active when they are in poor body condition. Social media can provide useful supplementary information for understanding of elusive mammals. These "soft" data can be applied to other species.

Finding biomarkers of experience in animals.

At a time when there is a growing public interest in animal welfare, it is critical to have objective means to assess the way that an animal experiences a situation. Objectivity is critical to ensure appropriate animal welfare outcomes. Existing behavioural, physiological, and neurobiological indicators that are used to assess animal welfare can verify the absence of extremely negative outcomes. But welfare is more than an absence of negative outcomes and an appropriate indicator should reflect the full spectrum of experience of an animal, from negative to positive. In this review, we draw from the knowledge of human biomedical science to propose a list of candidate biological markers (biomarkers) that should reflect the experiential state of non-human animals. The proposed biomarkers can be classified on their main function as endocrine, oxidative stress, non-coding molecular, and thermobiological markers. We also discuss practical challenges that must be addressed before any of these biomarkers can become useful to assess the experience of an animal in real-life.

Effect of Neck-Deep Immersion in Cool or Thermoneutral Water on Blood Glucose Levels in Individuals With Type 1 Diabetes.

Context: It is unclear whether immersion in cool water, typical of many beaches, increases the concentration of blood glucose in individuals with type 1 diabetes mellitus (T1DM). Objective: To test the hypothesis in individuals with T1DM that immersion neck-deep in cool water (COOL) causes an increase in blood glucose concentration, but not exposure to thermoneutral water (THERMO) or thermoneutral air. Methods: Eight overnight-fasted participants with T1DM were exposed for 60 minutes on separate days to 3 experimental conditions: cool water (COOL, 23 °C); thermoneutral water (THERMO, 33.5 °C); or thermoneutral air (24 °C). They then recovered for 60 minutes on land at 24 °C. At time intervals, we measured: blood glucose and plasma insulin concentration, rate of carbohydrate and fat oxidation, skin and core temperature, subcutaneous blood flow, and shivering via electromyography. Results: There was no change in blood glucose concentration during the 3 experimental conditions (P > .05). During recovery after COOL, blood glucose increased (P < .05) but did not change in the other 2 conditions. The rate of carbohydrate oxidation during and early after COOL was higher than in the other 2 conditions (P < .05), and COOL led to a decrease in subcutaneous blood flow and the concentration of plasma insulin (P < .05). Conclusion: Cool or thermoneutral neck-deep immersion in water does not cause a change in the concentration of blood glucose in people with T1DM, but on-land recovery from COOL causes an increase in blood glucose that may be due, at least in part, to the accompanying decrease in plasma insulin.

Correction: Could the ketogenic diet induce a shift in thyroid function and support a metabolic advantage in healthy participants? A pilot randomized-controlled-crossover trial.

[This corrects the article DOI: 10.1371/journal.pone.0269440.].

Cold and Hungry: Heterothermy Is Associated with Low Leptin Levels in a Bulk Grazer during a Drought.

AbstractReduced energy intake can compromise the ability of a mammal to maintain body temperature within a narrow 24-h range, leading to heterothermy. To investigate the main drivers of heterothermy in a bulk grazer, we compared abdominal temperature, body mass, body condition index, and serum leptin levels in 11 subadult Cape buffalo (Syncerus caffer caffer) during a drought year and a nondrought year. Low food availability during the drought year (as indexed by grass biomass, satellite imagery of vegetation greenness, and fecal chlorophyll) resulted in lower body condition index, lower body mass relative to that expected for an equivalent-aged buffalo, and lower leptin levels. The range of 24-h body temperature rhythm was 2°C during the nondrought year and more than double that during the drought year, and this was caused primarily by a lower minimum 24-h body temperature rhythm during the cool dry winter months. After rain fell and vegetation greenness increased, the minimum 24-h body temperature rhythm increased, and the range of 24-h body temperature rhythm was smaller than 2°C. In order of importance, poor body condition, low minimum 24-h air temperature, and low serum leptin levels were the best predictors of the increase in the range of 24-h body temperature rhythm. While the thermoregulatory role of leptin is not fully understood, the association between range of 24-h body temperature rhythm and serum leptin levels provides clues about the underlying mechanism behind the increased heterothermy in large mammals facing food restriction.

Pekin ducks are motivated to lay in their preferred nest substrate.

Nest design is one factor contributing to floor-laying in farmed poultry. We investigated: (i) if ducks (Anas platyrhynchos) prefer a particular nest substrate; and (ii) how important that preference is to them, indicated by stress-induced hyperthermia, egg albumen corticosterone, and behaviour. Twelve female ducks that were trained in a push-door task had temperature data loggers implanted. Preference testing identified the most and least preferred nest substrates between sawdust, astroturf, and hemp fibres. A behavioural demand test then required the ducks to use push-doors to access nests containing either the most or least preferred substrate. The preferred substrate door was loaded with increasing weight (0-120% of bodyweight, four nights per workload) and eventually blocked to prevent nest access. The least preferred substrate door remained unweighted. The overall rank order of substrate preferences was sawdust > hemp > astroturf. Six of the 12 birds pushed all workloads and attempted to push the blocked door. The area under the curve (AUC) of hyperthermia was larger when the preferred substrate door was blocked compared with 0%. The AUC did not differ between nights 2-4 of the blocked door compared with night 1. Egg albumen corticosterone was unaffected. We conclude that laying Pekin ducks prefer manipulatable nest substrates and accessing one is important enough to pay a cost. The results indicate that a manipulatable substrate should be provided to commercially farmed nesting ducks.

Could the ketogenic diet induce a shift in thyroid function and support a metabolic advantage in healthy participants? A pilot randomized-controlled-crossover trial.

BACKGROUND: The ketogenic diet (KD) has been shown to result in body mass loss in people with disease as well as healthy people, yet the effect of the KD on thyroid function and metabolism are unknown. OBJECTIVE: We aimed to determine the effects of a KD, compared with an isocaloric high-carbohydrate low-fat (HCLF) diet, on resting metabolic rate and thyroid function in healthy individuals. DESIGN: Eleven healthy, normal-weight participants (mean(SD) age: 30(9) years) completed this randomized crossover-controlled study. For a minimum of three weeks on each, participants followed two isocaloric diets: a HCLF diet (55%carbohydrate, 20%fat, 25%protein) and a KD (15%carbohydrate, 60%fat, 25% protein), with a one-week washout period in-between. Importantly, while on the KD, the participants were required to remain in a state of nutritional ketosis for three consecutive weeks. Crossover analyses and linear mixed models were used to assess effect of diet on body mass, thyroid function and resting metabolic rate. RESULTS: Both dietary interventions resulted in significant body mass loss (p<0.05) however three weeks of sustained ketosis (KD) resulted in a greater loss of body mass (mean (95%CI): -2.9 (-3.5, -2.4) kg) than did three weeks on the HCLF diet (-0.4 (-1.0, 0.1) kg, p < 0.0001). Compared to pre-diet levels, the change in plasma T3 concentration was significantly different between the two diets (p = 0.003), such that plasma T3 concentration was significantly lower following the KD diet (4.1 (3.8, 4.4) pmol/L, p<0.0001) but not different following the HCLF diet (4.8 (4.5, 5.2) pmol/L, p = 0.171. There was a significant increase in T4 concentration from pre-diet levels following the KD diet (19.3 (17.8, 20.9) pmol/L, p < 0.0001), but not following the HCLF diet (17.3 (15.7, 18.8) pmol.L, p = 0.28). The magnitude of change in plasma T4 concentration was not different between the two diets (p = 0.4). There was no effect of diet on plasma thyroid stimulating hormone concentration (p = 0.27). There was a significantly greater T3:T4 ratio following the HCLF diet (0.41 (0.27, 0.55), p < 0.0001) compared to pre-diet levels but not following the KD diet (0.25 (0.12, 0.39), p = 0.80). CONCLUSIONS: Although the diets were isocaloric and physical activity and resting metabolic rate remained constant, the participants lost more mass after the KD than after the HCLF diet. The observed significant changes in triiodothyronine concentration suggest that unknown metabolic changes occur in nutritional ketosis, changes that warrant further investigation. TRIAL REGISTRATION: Pan African Clinical Trial Registry: PACTR201707002406306 URL: https://pactr.samrc.ac.za/.

Effects of Simulated High Altitude on Blood Glucose Levels During Exercise in Individuals With Type 1 Diabetes.

CONTEXT: Current exercise guidelines for individuals with type 1 diabetes (T1D) do not consider the impact that high altitude may have on blood glucose levels (BGL) during exercise. OBJECTIVE: To investigate the effect of acute hypoxia (simulated high altitude) on BGL and carbohydrate oxidation rates during moderate intensity exercise in individuals with T1D. METHODS: Using a counterbalanced, repeated measures study design, 7 individuals with T1D completed 2 exercise sessions; normoxia and hypoxia (~4200 m simulated altitude). Participants cycled for 60 min on an ergometer at 45% of their sea-level V̇O2peak, and then recovered for 60 min. Before, during, and after exercise, blood samples were taken to measure glucose, lactate, and insulin levels. Respiratory gases were collected to measure carbohydrate oxidation rates. RESULTS: Early during exercise (<30 min), there was no fall in BGL in either condition. After 1 h of exercise and during recovery, BGL were significantly lower under the hypoxic condition compared to both pre-exercise levels (P = 0.008) and the normoxic condition (P = 0.027). Exercise in both conditions resulted in a significant rise in carbohydrate oxidation rates, which returned to baseline levels postexercise. Before, during, and after exercise, carbohydrate oxidation rates were higher under the hypoxic compared with the normoxic condition (P < 0.001). CONCLUSIONS: The greater decline in BGL during and after exercise performed under acute hypoxia suggests that exercise during acute exposure to high altitude may increase the risk of hypoglycemia in individuals with T1D. Future guidelines may have to consider the impact altitude has on exercise-mediated hypoglycemia.

Kisspeptin impacts on circadian and ultradian rhythms of core body temperature: Evidence in kisspeptin receptor knockout and kisspeptin knockdown mice.

Kisspeptin is vital for the regulation of both fertility and metabolism. Kisspeptin receptor (Kiss1r) knockout (KO) mice exhibit increased adiposity and reduced energy expenditure in adulthood. Kiss1r mRNA is expressed in brown adipose tissue (BAT) and Kiss1r KO mice exhibit reduced Ucp1 mRNA in BAT and impaired thermogenesis. We hypothesised that mice with diminished kisspeptin signalling would exhibit reduced core body temperature (Tc) and altered dynamics of circadian and ultradian rhythms of Tc. Tc was recorded every 15-min over 14-days in gonadectomised wild-type (WT), Kiss1r KO, and also Kiss1-Cre (95% reduction in Kiss1 transcription) mice. Female Kiss1r KOs had higher adiposity and lower Ucp1 mRNA in BAT than WTs. No change was detected in Kiss1-Cre mice. Mean Tc during the dark phase was lower in female Kiss1r KOs versus WTs, but not Kiss1-Cre mice. Female Kiss1r KOs had a lower mesor and amplitude of the circadian rhythm of Tc than did WTs. In WT mice, there were more episodic ultradian events (EUEs) of Tc during the dark phase than the light phase, but this measure was similar between dark and light phases in Kiss1r KO and Kiss1-Cre mice. The amplitude of EUEs was higher in the dark phase in female Kiss1r KO and male Kiss1-Cre mice. Given the lack of clear metabolic phenotype in Kiss1-Cre mice, 5% of Kiss1 transcription may be sufficient for proper metabolic control, as was shown for fertility. Moreover, the observed alterations in Tc suggest that kisspeptin has a role in circadian and ultradian rhythm-driven pathways.

Diet-altered body temperature rhythms are associated with altered rhythms of clock gene expression in peripheral tissues in vivo.

Temperature rhythms can act as potent signals for the modulation of the amplitude and phase of clock gene expression in peripheral organs in vitro, but the relevance of the circadian rhythm of core body temperature (Tc) as a modulating signal in vivo has not yet been investigated. Using calorie restriction and cafeteria feeding, we induced a larger and a dampened Tc amplitude, respectively, in male Wistar rats, and investigated the circadian expression profile of the core clock genes Bmal1, Per2, Cry1, and Rev-erbα, the heat-responsive genes heat shock protein 90 (Hsp90) and cold-inducible RNA binding protein (Cirbp), and Pgc1α, Pparα/γ/δ, Glut1/4, and Chop10 in the liver, skeletal muscle, white adipose tissue (WAT), and adrenal glands. Diet-altered Tc rhythms differentially affected the profiles of clock genes, Hsp90, and Cirbp expression in peripheral tissues. Greater Tc amplitudes elicited by calorie restriction were associated with large amplitudes of Hsp90 and Cirbp expression in the liver and WAT, in which larger amplitudes of clock gene expression were also observed. The amplitudes of metabolic gene expression were greater in the WAT, but not in the liver, in calorie-restricted rats. Conversely, diet-altered Tc rhythms were not translated to distinct changes in the amplitude of Hsp90, Cirbp, or clock or metabolic genes in the skeletal muscle or adrenal glands. While it was not possible to disentangle the effects of diet and temperature in this model, taken together with previous in vitro studies, our study presents novel data consistent with the notion that the circadian Tc rhythm can modulate the amplitude of circadian gene expression in vivo. The different responses of Hsp90 and Cirbp in peripheral tissues may be linked to the tissue-specific responses of peripheral clocks to diet and/or body temperature rhythms, but the association with the amplitude of metabolic gene expression is limited to the WAT.

Comparative Expression Profiling and Sequence Characterization of ATP1A1 Gene Associated with Heat Tolerance in Tropically Adapted Cattle.

Climate change is an imminent threat to livestock production. One adaptation strategy is selection for heat tolerance. While it is established that the ATP1A1 gene and its product play an important role in the response to many stressors, there has been no attempt to characterize the sequence or to perform expression profiling of the gene in production animals. We undertook a field experiment to compare the expression profiles of ATP1A1 in heat-tolerant Vechur and Kasaragod cattle (Bos taurus indicus) with the profile of a heat-susceptible crossbreed (B. t. taurus × B. t. indicus). The cattle were exposed to heat stress while on pasture in the hot summer season. The environmental stress was quantified using the temperature humidity index (THI), while the heat tolerance of each breed was assessed using a heat tolerance coefficient (HTC). The ATP1A1 mRNA of Vechur cattle was amplified from cDNA and sequenced. The HTC varied significantly between the breeds and with time-of-day (p < 0.01). The breed-time-of-day interaction was also significant (p < 0.01). The relative expression of ATP1A1 differed between heat-tolerant and heat-susceptible breeds (p = 0.02). The expression of ATP1A1 at 08:00, 10:00 and 12:00, and the breed-time-of-day interaction, were not significant. The nucleotide sequence of Vechur ATP1A1 showed 99% homology with the B. t. taurus sequence. The protein sequence showed 98% homology with B. t. taurus cattle and with B. grunniens (yak) and 97.7% homology with Ovis aries (sheep). A molecular clock analysis revealed evidence of divergent adaptive evolution of the ATP1A1 gene favoring climate resilience in Vechur cattle. These findings further our knowledge of the relationship between the ATP1A1 gene and heat tolerance in phenotypically incongruent animals. We propose that ATP1A1 could be used in marker assisted selection (MAS) for heat tolerance.

A Comparison of Physical Activity Levels, Sleep Disrupting Behavior, and Stress/Affective Distress as Predictors of Sleep as Indexed by Actigraphy.

BACKGROUND: Several behaviors have been reported to interfere with sleep in otherwise healthy adults, including low physical activity (PA) levels. However, few studies have compared low PA with the other behavioral risk factors of objective sleep impairment, despite the behavior tending to cooccur in highly stressed and affectively distressed individuals. Thus, the authors compared objective and subjective measures of PA and other potential sleep disrupting behaviors as predictors of objective sleep (sleep onset latency, actual sleep time, total sleep duration, awake time, and sleep efficacy) at baseline (T1) and 3 months later (T2). METHODS: A community-derived sample of 161 people aged 18-65 years were asked about PA, other behavior (ie, night eating, electronic device use, watching television, caffeine and alcohol use), stress, affective distress (ie, anxiety, depression), and demographics including shift work and parenting young children in an online questionnaire at T1 and T2. PA and sleep were also monitored for 24 hours each at T1 and T2 using actigraphy. RESULTS: Multiple regression analyses indicated that sleep at T1 was associated with PA (ie, total number of steps, metabolic equivalents/time, time spent travelling) after controlling mean ambient temperature and relevant demographics. At T2, longer sleep onset latency was predicted by parenting young children and night time television viewing; shorter sleep duration was predicted by female gender; and awake time and sleep efficacy were predicted by alcohol intake after controlling T1 sleep measures, demographics, and mean ambient temperature. CONCLUSION: The risk factors for objective sleep impairment included parenting young children and watching television at night, whereas better sleep outcomes were associated with greater engagement with PA.

Daily temperature cycles prolong lifespan and have sex-specific effects on peripheral clock gene expression in Drosophila melanogaster.

Circadian rhythms optimize health by coordinating the timing of physiological processes to match predictable daily environmental challenges. The circadian rhythm of body temperature is thought to be an important modulator of molecular clocks in peripheral tissues, but how daily temperature cycles affect physiological function is unclear. Here, we examined the effect of constant temperature (Tcon, 25°C) and cycling temperature (Tcyc, 28°C:22°C during light:dark) paradigms on lifespan of Drosophila melanogaster, and the expression of clock genes, heat shock protein 83 (Hsp83), Frost (Fst) and senescence marker protein-30 (smp-30). Male and female D. melanogaster housed at Tcyc had longer median lifespans than those housed at Tcon. Tcyc induced robust Hsp83 rhythms and rescued the age-related decrease in smp-30 expression that was observed in flies at Tcon, potentially indicating an increased capacity to cope with age-related cellular stress. Ageing under Tcon led to a decrease in the amplitude of expression of all clock genes in the bodies of male flies, except for cyc, which was non-rhythmic, and for per and cry in female flies. Strikingly, housing under Tcyc conditions rescued the age-related decrease in amplitude of all clock genes, and generated rhythmicity in cyc expression, in the male flies, but not the female flies. The results suggest that ambient temperature rhythms modulate D. melanogaster lifespan, and that the amplitude of clock gene expression in peripheral body clocks may be a potential link between temperature rhythms and longevity in male D. melanogaster. Longevity due to Tcyc appeared predominantly independent of clock gene amplitude in female D. melanogaster.

How dryland mammals will respond to climate change: the effects of body size, heat load and a lack of food and water.

Mammals in drylands are facing not only increasing heat loads but also reduced water and food availability as a result of climate change. Insufficient water results in suppression of evaporative cooling and therefore increases in body core temperature on hot days, while lack of food reduces the capacity to maintain body core temperature on cold nights. Both food and water shortage will narrow the prescriptive zone, the ambient temperature range over which body core temperature is held relatively constant, which will lead to increased risk of physiological malfunction and death. Behavioural modifications, such as shifting activity between night and day or seeking thermally buffered microclimates, may allow individuals to remain within the prescriptive zone, but can incur costs, such as reduced foraging or increased competition or predation, with consequences for fitness. Body size will play a major role in predicting response patterns, but identifying all the factors that will contribute to how well dryland mammals facing water and food shortage will cope with increasing heat loads requires a better understanding of the sensitivities and responses of mammals exposed to the direct and indirect effects of climate change.

Calm Hu ram lambs assigned by temperament classification are healthier and have better meat quality than nervous Hu ram lambs.

The objective of this study was to evaluate the effects of temperament classification (assessed using an arena test) on health and productivity of Hu ram lambs. In experiment one, eight ram lambs classified as calm and eight classified as nervous (selected from 100 ram lambs) were fed individually for 60-days to compare food intake, food digestibility, weight gain, and biochemical indices of health. In experiment two, nine ram lambs classified as calm and nine classified as nervous (selected from 150 ram lambs) were fed in a group and slaughter traits, meat quality, and muscle histology were compared. Calm lambs had higher dry matter digestibility, lower serum TNF-α, higher total antioxidant capacity, higher total superoxide dismutase activity, higher dressing percentage, higher cross-sectional area of loin, higher myofibre density, lower ultimate pH of the meat, and higher meat redness, than nervous lambs. Selection for calm temperament could be beneficial to health, slaughter, and carcass traits in Hu ram lambs.

Pekin ducks are motivated to access their nest site and exhibit a stress-induced hyperthermia when unable to do so.

The origins of floor-laying in ducks could be low motivation for a nest, or stress related to difficulties with accessing a nest (e.g. competition). Using a behavioural demand test, we investigated if increasing the work required to access their nest impacted ducks' behaviour and two indicators of stress: egg corticosterone concentration and elevation of core body temperature (stress-induced hyperthermia (SIH)). Twelve laying Pekin ducks previously trained in an operant push-door task were required to use a push-door to access their nest. The door was loaded with increasing weight (0-160% of individual BW, four nights per workload) and eventually blocked to prevent nest access. Before testing, temperature data loggers were implanted in the abdomen. Eggs were collected daily to measure corticosterone concentrations. Behaviour towards the push-door was quantified. Three birds were excluded from the experiment at an early stage. Five of the nine remaining birds pushed all workloads up to 160% BW and attempted to pass the blocked door, with another two birds pushing up to 80 and 140% BW. For those that pushed at all workloads (n = 5) the area under the curve (AUC) of hyperthermia was larger at workloads of 80% (P < 0.001), 120% (P < 0.01), 140% (P < 0.001), 160% (P < 0.001), and when the door was blocked (P < 0.001), compared with 0%. On the first night when the door was blocked, all five birds pushed more at the door, but no attempts were made to push on the following 3 nights, yet the AUC of hyperthermia did not differ between nights 2-4 of the blocked door, compared with the first night that the door was blocked. Increasing workload and inability to access the nest had no effect on corticosterone in egg albumen. It was concluded that laying Pekin ducks were motivated to access a nest. Although it was not possible to differentiate metabolic from psychogenic stress on the first night that nest access was denied, we suggest that the occurrence of hyperthermia on the subsequent nights was due to SIH resulting from frustration at their inability to use their preferred nest. Floor-laying therefore is unlikely due solely to low nest-seeking motivation. Egg corticosterone was not a relevant indicator of acute stress. Strategies to improve nest availability (e.g. decreasing competition) could improve the welfare of commercial ducks.