Loss of a grooming enrichment impacts physical, behavioural, and physiological measures of welfare in grazing beef cattle.

Pasture-based beef cattle are raised in a range of production environments. Some paddocks may contain trees and other objects that allow for grooming, hence being naturally enriching, whilst others may be barren without these opportunities. Additionally, it is not uncommon for cattle to move between these enriched and barren environments as part of routine management. While the benefits of enrichment are well studied, how this 'enrichment loss' impacts cattle welfare as access to stimuli is removed is unknown. This trial assessed the impacts of the loss of an enriching object (grooming brush) on grazing beef cattle welfare and production characteristics. When grooming brush access was blocked, cattle became dirtier, showed reduced average daily gain, and had elevated faecal cortisol metabolites, although this varied according to the degree of initial individual brush use. Additionally, allogrooming and grooming on other objects were reduced when access to the brush was returned, potentially indicating a rebound effect. These results demonstrate that the loss of adequate grooming objects can impair the overall welfare of grazing cattle; however, further work is needed to determine exactly which natural or artificial objects provide adequate grooming opportunities.

Sex impacts pain behaviour but not emotional reactivity of lambs following ring tail docking.

Studies in humans have shown sex differences in response to painful events, however, little is known in relation to sex differences in sheep. Understanding sex differences would enable improved experimental design and interpretation of studies of painful procedures in sheep. To examine sex differences in response to pain, 80 lambs were tested across five cohorts of 16. The lambs were penned in groups containing two male and two female lambs with their respective mothers. Lambs were randomly allocated from within each block to one of four treatment groups; FRing-Female lamb, ring tail docked without analgesia, MRing-Male lamb, ring tail docked without analgesia, FSham-Female lamb, tail manipulated and MSham-Male lamb, tail manipulated. Following treatment, lambs were returned to their pen and were video recorded for 45 mins for behavioural observations of acute pain and posture. An hour after treatment, lambs then underwent an emotional reactivity test that consisted of three phases: Isolation, Novelty and Startle. Following treatment, Ring lambs displayed more abnormal postures (mean = 2.5 ± 0.5) compared to Sham lambs (mean = 0.05 ± 0.4, P = 0.0001). There was an effect of sex on the display of acute pain-related behaviours in lambs that were tail docked (P < 0.001), with female lambs displaying more acute behaviours (mean count = +2.2). This difference in behaviour between sexes was not observed in Sham lambs. There was no effect of sex on display of postures related to pain (P = 0.99). During the Novelty and Startle phase of the emotional reactivity test, Ring lambs tended to (P = 0.084) or did (P = 0.018) show more fear related behaviours, respectively. However, no effect of sex was observed. The results of this study indicate that a pain state may alter the emotional response of lambs to novel objects and potential fearful situations. It was also demonstrated that female lambs display increased sensitivity to the acute pain caused by tail docking compared to males.

Repeatability of an attention bias test for sheep suggests variable influence of state and trait affect on behaviour.

Understanding the effects of repeated testing on behaviour is essential for behavioural tests that are re-applied to the same individuals for research and welfare assessment purposes. Assessing the repeatability of behaviour can also help us understand the influence of persistent traits vs transient states on animal responses during testing. This study examined the repeatability of behavioural responses in an attention bias test developed for sheep as a measure of affective state. Sheep were assessed in the attention bias test three times (n = 81 sheep), with testing occurring at intervals of 1 year then 2 weeks. During testing, individual sheep were exposed to a dog located behind a window for 3 s in a 4 × 4 m arena, then the dog was obscured from view, removed and sheep behaviours were recorded for 180 s. We hypothesised that behaviours in the test would have moderate-high repeatability but that the mean behavioural responses would change over consecutive trials as sheep habituated to the test environment. To estimate repeatability, data were modelled using restricted maximum likelihood linear mixed-effects models, fitting animal ID as a random effect. Vigilance behaviour, defined as having the head at or above shoulder height, was moderately repeatable (r = 0.58). Latency to eat (r = 0.20) and duration spent looking towards the previous location of the dog (attention to the dog wall) (r = 0.08) had low repeatability. Mean latency to eat did not differ significantly between trials (P = 0.2) and mean vigilance behaviour tended to decrease over the trials (P = 0.07). Mean duration of attention to the dog wall significantly decreased across the trials (P < 0.001), while mean zones crossed increased (P < 0.001), as did behaviours directed towards the exit door such as duration in proximity and pawing at the door. Overall, vigilance behaviour was moderately repeatable, suggesting it may have been driven by temperament or personality traits, while attention and feeding behaviours may have been more influenced by transient affective states or other factors, however further research is needed to better tease apart these potential effects. Sheep demonstrated some habituation to the test over consecutive trials. Care should therefore be taken during future application of the test to ensure all animals undergoing attention bias testing have equivalent experience for a valid interpretation of their relative behavioural responses.

Beef Cattle Preference and Usage of Environmental Enrichments Provided Simultaneously in a Pasture-Based Environment.

Environmental enrichment can improve livestock welfare through increasing environmental complexity to promote a greater range of natural behaviours. However, there is limited understanding of the need for and impacts of enrichments for extensively managed beef cattle that can sometimes be kept in grassed paddocks devoid of additional natural and artificial features, i.e., 'barren pastures'. This trial assessed which enrichments beef cattle preferred and utilised in a barren paddock environment. Eight groups of seven Angus steers housed on pastured paddocks devoid of natural or artificial features were observed during daylight hours for two days a week over a period of three weeks, after being presented with four enrichments simultaneously: a cattle brush, a piece of hanging rope, a tree stump, and a woodchip pile. Although enrichment use generally decreased over time, the brush, stump, and woodchip maintained a higher level of use than the rope, based on the frequency of interactions and number of displacements around the enrichments (both p < 0.001). This suggests that the brush, stump, and woodchip pile were more valuable resources to the cattle, allowing for grooming and lying behaviours, although oral manipulations also occurred on the stump, woodchip, and rope. The inclusion of these enrichments can increase the complexity of barren pasture environments and allow for the increased expression of natural behaviours, potentially contributing to improved welfare.

Attention Bias Test Measures Negative But Not Positive Affect in Sheep: A Replication Study.

An attention bias test has been developed as a measure of negative affective states in sheep. The test measures an individual's allocation of attention between a threatening (previous location of a dog) and positive (conspecific photo) stimulus over a 3 min period. This study replicated a previously inconclusive study, to determine whether the test could assess positive affective states under more controlled conditions and with a younger population of animals. Pharmacological treatments were used to induce anxious, calm, happy, and control affective states prior to entering the attention bias test arena (n = 20/treatment). We hypothesized that sheep in positive and negative affective states could be differentiated using key measures of attention during testing, including vigilance (head at or above shoulder height) and duration looking towards the valenced stimuli. Anxious sheep were more vigilant than control animals during attention bias testing as predicted (linear mixed effects model, p = 0.002), but the positive groups did not differ from controls (p > 0.05). There was no effect of treatment on looking behaviors (p > 0.05). We suggest this attention bias test paradigm can assess negative but not positive affect in sheep and that modifications to the ethogram or stimuli are needed to more clearly characterize the direction of attention during testing.

Pharmacologically-induced stress has minimal impact on judgement and attention biases in sheep.

The emotional impact of exposure to stressors has not been well quantified in animals. We hypothesised that exogenous induction of stress in sheep would induce a pessimistic judgement bias and increased attention towards a threatening stimulus, suggestive of a negative emotional state. Stress was induced pharmacologically by administering synthetic adrenocorticotropic hormone. Judgement bias was assessed using a spatial go/no-go task after exposure to acute stress (one injection), chronic stress (21 daily injections) and acute-on-chronic stress (2 min isolation after 28 daily injections). Attention bias was assessed during chronic stress only (22 daily injections). In contrast with our hypotheses, there was no strong evidence that Synacthen administration altered judgement bias or attention bias at any stage of the experiment. Stressed sheep were more likely to approach ambiguous locations than saline Control animals, however, statistical evidence for models fitting treatment group was very weak. Overall, our findings suggest that elevated levels of cortisol may not fully explain changes to judgement bias observed in previous studies after environmentally-induced stress. Further studies are required to better understand which aspects of environmentally-induced stress alter judgement bias and to further validate cognitive methods of assessing affect in sheep.

The influence of pharmacologically-induced affective states on attention bias in sheep.

When an individual attends to certain types of information more than others, the behavior is termed an attention bias. The occurrence of attention biases in humans and animals can depend on their affective states. Based on evidence from the human literature and prior studies in sheep, we hypothesized that an attention bias test could discriminate between pharmacologically-induced positive and negative affective states in sheep. The test measured allocation of attention between a threat and a positive stimulus using key measures of looking time and vigilance. Eighty 7-year-old Merino ewes were allocated to one of four treatment groups; Anxious (m-chlorophenylpiperazine), Calm (diazepam), Happy (morphine) and Control (saline). Drugs were administered 30 min prior to attention bias testing. The test was conducted in a 4 × 4.2 m arena with high opaque walls. An approximately life-size photograph of a sheep was positioned on one wall of the arena (positive stimulus). A small window with a retractable opaque cover was positioned on the opposite wall, behind which a dog was standing quietly (threat). The dog was visible for 3 s after a single sheep entered the arena, then the window was covered and the dog was removed. Sheep then remained in the arena for 3 min while behaviors were recorded. Key behaviors included time looking toward the dog wall or photo wall, duration of vigilance behavior and latency to become non-vigilant. In contrast with our hypothesis, no significant differences were found between treatment groups for duration of vigilance or looking behaviors, although Anxious sheep tended to be more vigilant than Control animals (P < 0.1) and had a longer latency to become non-vigilant (P < 0.001). A total of 24 of 80 animals were vigilant for the entire test duration. This censoring of data may explain why no differences were detected between groups for vigilance duration. Overall, a lack of difference between groups may suggest the test cannot discriminate positive and negative states in sheep. We suggest that the censoring of vigilance duration data, the use of insufficient drug doses, the potential influence of background noise and the age of the sheep may explain a lack of difference between groups. Due to these potential effects, it remains unclear whether the attention bias test can detect positive states in sheep.

Attention Bias Test Differentiates Anxiety and Depression in Sheep.

Negative affective states such as anxiety and depression pose a risk to animal welfare, however, practical tests for assessing these states in animals are limited. In humans, anxious individuals are shown to pay more attention toward threatening information than non-anxious individuals, known as an attention bias. Previously, an attention bias test was developed and validated as a measure of anxious states in sheep, where more anxious sheep showed increased attention toward a threat (dog) and were more vigilant than Control animals. Studies in humans suggest that attention biases also occur in depressed individuals, with observations of attention biases toward threats, as well as biases away from positive stimuli. Given these findings, we hypothesized that an attention bias test for sheep could also be used to assess states of depression. We predicted that Merino ewes in pharmacologically induced Depressed (para-chlorophenylalanine) and Anxious (m-chlorophenylpiperazine) states would show greater attention toward a threat than Control animals (saline), but that the Depressed sheep would show relatively less interest in a positive stimulus (photograph of a conspecific). During testing, Depressed sheep paid more attention toward the threat and less toward the photograph than Control animals as predicted (Analyses of Variance, P < 0.05, n = 16 per treatment). Interestingly, Anxious sheep showed an attention bias in the opposite direction, paying more attention toward the photograph and less toward the threat than Control animals (P < 0.05). Both Anxious and Depressed sheep were more vigilant than Control animals (P = 0.002). These results suggest the attention bias test can be used to measure and differentiate states of depression and anxiety in livestock. The bidirectional nature of the attention bias identified between treatments highlights the importance of measuring multiple behaviors in the test and considering the context in which the test is applied. This will enable a clearer characterization of the affective state of an animal, as an aspect of its welfare.

A Novel Protocol to Assess Acclimation Rate in Bos taurus Heifers during Yard Weaning.

The speed with which animals acclimate to a new environment could be an important measure of ability to cope with management induced stress. This study developed a measure of acclimation rate in a group of 50 Bos taurus heifers during yard weaning over nine days. We recorded the time and order in which heifers moved through a novel funnel structure into a feeding yard daily. We hypothesised that addition of an obstacle at the entrance would increase the time it took heifers to move through the funnel, but that they would acclimate to the obstacle over a three-day period. The change in latency to move through could then be used as a measure of acclimation rate. We hypothesised that individuals which acclimated to obstacles at a faster rate might display favourable temperament as assessed by flight time. All heifers took longer to move through the funnel after a novel object was introduced, then latency decreased over the following two days while the object was present. This indicates the protocol could be useful for measuring acclimation rate at a group level. Individual acclimation rate variables, measured as change in times and orders of heifers between test days, did not appear to have any consistent relationships with flight time or weight change during or post-weaning (p > 0.05). We concluded that the protocol was inappropriate for assessing acclimation rate at an individual level, due to social effects while testing heifers as a group. Heifers which were consistently one of the first 20 to move through the funnel had a significantly greater average weight 5 and 10 months post-weaning (345 ± 9 kg and 518 ± 10 kg respectively) than heifers which were consistently one of the last 20 through the funnel (311 ± 8 kg and 484 ± 8 kg respectively; p < 0.001). This may indicate order of movement through the funnel was related to feeding motivation or another aspect of temperament not reflected by flight time.

Towards a more practical attention bias test to assess affective state in sheep.

Tests for attention bias potentially offer more rapid assessment of affective state in animals than existing cognitive methods. An attention bias test has previously been developed for sheep and validated as a measure of anxious states. The 3 minute test assessed behavioural responses of sheep in an enclosed arena after brief exposure to the threat of a dog. Experiment 1 of the current study aimed to refine the previously developed method, removing the need for a habituation period and shortening the test duration. Sheep were given either an anxiolytic drug, an anxiogenic drug or a control treatment prior to testing to induce contrasting affective states. Differences in behaviour were found between the treatment groups within the first 45s of the test, indicating the original test duration could be shortened from 180 s. During testing, 36 of 40 animals in the control and anxiolytic groups ate the novel feed offered in the test, indicating it is not necessary to habituate animals to a feed container. Experiment 2 aimed to confirm the responses measured in the test were primarily towards the dog rather than other aspects of the test environment. Sheep exposed to an empty window at the beginning of the test behaved differently to those which were exposed to a dog, indicating sheep behaviour in the test is at least partially a response to the dog. A third group of sheep were also tested with the dog immediately after having small data loggers attached to their necks. Behaviour of these sheep did not differ from the sheep tested without loggers, indicating data logger attachment did not impact their behaviour in the test. In both experiments, treatments did not appear to modify activity (zones crossed), which we propose indicates the test was primarily detecting valence of the affective state rather than arousal.