Animal Welfare Guidelines for International Development Organisations in the Global South.

International development organisations have improved billions of human lives in the Global South. However, in both their projects and advice to governments, most of these organisations neglect animal welfare. This blindspot matters. Poor welfare standards risk the organisation's reputation, particularly with donors; they reduce livestock lifespans and productivity, harming recipients; and they cause animals unnecessary pain and suffering. Here, we set out animal welfare guidelines for international development organisations. They were developed through extensive stakeholder engagement with organisations, donors, and recipients, especially in Africa. To comprehensively cover animal welfare, the guidelines encompass governance structure within the organisation, staff training, standard operating procedures, water, food, housing, social isolation, enrichment, drainage and waste disposal, disease, invasive procedures, transport, slaughter, breeds, record-keeping, and monitoring and evaluation of success. We urge international development organisations to adopt and institutionalise these guidelines, so they promote good animal welfare.

Is it time for insect researchers to consider their subjects' welfare?

Recent evidence suggests that at least some insect species might plausibly feel pain. These findings should prompt researchers to think about the welfare implications of insect experiments.

Short-Term Microplastic Exposure Impairs Cognition in Hermit Crabs.

We tested whether acute microplastic exposure impacts information gathering and processing (cognition) in hermit crabs (Pagurus bernhardus). For five days, we kept 51 hermit crabs in tanks containing either polyethylene microspheres (n = 27) or no plastic (n = 24). We then transferred individuals into an intermediate-quality shell and presented them with two vials containing either a better or worse shell. Because touching both shell vials required an equivalent behavioural response, this design controlled for general activity. Plastic-exposed hermit crabs were less likely and slower than controls to touch the better shell vial, instead preferring the worse shell vial. Microplastics, therefore, impaired assessments and decision-making, providing direct evidence of acute microplastic exposure disrupting hermit crab cognition.

Does microplastic exposure and sex influence shell selection and motivation in the common European hermit crab, Pagurus bernhardus?

Microplastics (<5 mm) are a threat to marine biodiversity however their effects on animal cognition and behaviour are unclear. We investigated whether microplastic exposure affects shell selection behaviour and motivation in the common European hermit crab, Pagurus bernhardus. Subjects were maintained for 5 days in tanks containing either: polyethylene microplastic spheres (n = 40), or no plastic (n = 40). They were then placed in low-quality shells and presented with an alternative high-quality shell. When they first touched the high-quality shell, the hermit crabs were startled using visual and aural stimuli. We recorded the post-startle latency to re-contact the high-quality shell, quantifying motivation to explore and acquire a better shell. Plastic-exposed females were more likely to select the high-quality shell than control females. As hypothesised, female hermit crabs had longer initial contact latencies, startle durations, and shell entry latencies than males. We also found an interaction effect on shell investigation duration: females from the control treatment spent longer investigating the high-quality shell compared to males. This was absent in the microplastic treatment with females behaving similar to males. This controlled study serves as a starting point to investigate the effects of microplastics and sex differences on behaviour when under predatory threat, and demonstrated sex dependent sensitivity to an environmental pollutant of global concern.

Canine Socialisation: A Narrative Systematic Review.

There are over 10 million pet dogs in the UK alone, and they have become a member of modern human families. If not properly socialised as puppies, dogs have a higher risk of problematic behaviours during adulthood, yet socialisation studies are lacking. Much of the experimental research was carried out at least 50 years ago, and the importance of socialisation was demonstrated so clearly that further studies with unsocialised controls would be deemed unethical. In this review, the aim was to evaluate all literature relevant to canine socialisation. This review used PRISMA-P guidelines to identify 29 studies: 14 were questionnaire-based studies (two of which also had a testing element), 15 included some form of experimental manipulation relating to socialisation, and one was a purely observational study. Based on this literature review, we recommend future research into minimum necessary socialisation levels, as well as breed differences in the timing of effective socialisation. Such studies will help owners and breeders produce well-adjusted adult dogs.

Motivational trade-offs and modulation of nociception in bumblebees.

Insects are traditionally thought to respond to noxious stimuli in an inflexible manner, without the ability to modulate their behavior according to context. We investigated whether bumblebees' attraction to high sucrose solution concentrations reduces their avoidance of noxious heat. Bees were given the choice between either unheated or noxiously heated (55 °C) feeders with different sucrose concentrations and marked by different colors. Bees avoided noxious feeders when the unheated feeders contained high sucrose concentrations, but progressively increased feeding from noxious feeders when the sucrose concentration at unheated feeders decreased. This shows a motivational trade-off of nociceptive responses. Bees used learned color cues for their decisions, and thus the trade-off was based on processing in the brain, rather than just peripheral processing. Therefore, bees can use contextual information to modulate nociceptive behavior. This ability is consistent with a capacity for pain experiences in insects.

Pasture access and eye temperature in dairy cows.

Pasture access can benefit dairy cows' behavior, health, and welfare, but herds are increasingly housed indoors full-time. Recent infrared thermal-imaging (thermography) studies suggest that higher eye temperatures may be a physiological indicator of chronic stress. We, therefore, hypothesized that, compared to cows with pasture access, cows housed indoors full-time would have higher eye temperatures. In a two-phase crossover experiment, 29 Holstein-Friesian dairy cows experienced 18 days of overnight pasture access and 18 days of full-time indoor housing. We measured each animal's eye temperature 16 times (eight/phase). During Phase One, cows with pasture access had higher eye temperatures than cows housed indoors full-time (contrary to our hypothesis). However, during Phase Two, cows with pasture access had lower eye temperatures than cows housed indoors full-time. It is, therefore, unclear whether eye temperature reflected disparities in dairy cow welfare between different housing treatments.

Correction: McEvoy et al. Canine Socialisation: A Narrative Systematic Review. Animals 2022, 12, 2895.

The authors wish to make the following corrections to this paper [...].

Separating conscious and unconscious perception in animals.

In a new study, Ben-Haim et al. use subliminal stimuli to separate conscious and unconscious perception in macaques. A programme of this type, using a range of cognitive tasks, is a promising way to look for conscious perception in more controversial cases.

Animal contests and microplastics: evidence of disrupted behaviour in hermit crabs Pagurus bernhardus.

Microplastics are ubiquitous in global marine systems and may have negative impacts on a vast range of species. Recently, microplastics were shown to impair shell selection assessments in hermit crabs, an essential behaviour for their survival. Hermit crabs also engage in 'rapping' contests over shells, based on cognitive assessments of shell quality and opponent fighting ability and, hence, are a useful model species for examining the effects of microplastics on fitness-relevant behaviour in marine systems. Here, we investigated how a 5-day microplastic exposure (25 microplastics/litre) affected the dynamics and outcome of 120 staged hermit crab contests. Using a 2 × 2 factorial design, we examined how microplastics (i.e. presence or absence) and contestant role (i.e. attacker or defender) affected various behavioural variables. Significantly higher raps per bout were needed to evict microplastic-treated defenders when attackers were pre-exposed to control conditions (i.e. no plastic). Also, significantly longer durations of rapping bouts were needed to evict control-treated defenders when attackers were pre-exposed to microplastics. We suggest that microplastics impaired defenders' ability to identify resource holding potential and also affected attackers' rapping strength and intensity during contests. These impacts on animal contests indicate that microplastics have broader deleterious effects on marine biota than currently recognized.

Effect of Environmental Complexity and Stocking Density on Fear and Anxiety in Broiler Chickens.

Barren housing and high stocking densities may contribute to negative affective states in broiler chickens, reducing their welfare. We investigated the effects of environmental complexity and stocking density on broilers' attention bias (measure of anxiety) and tonic immobility (measure of fear). In Experiment 1, individual birds were tested for attention bias (n = 60) and in Experiment 2, groups of three birds were tested (n = 144). Tonic immobility testing was performed on days 12 and 26 (n = 36) in Experiment 1, and on day 19 (n = 72) in Experiment 2. In Experiment 1, no differences were observed in the attention bias test. In Experiment 2, birds from high-complexity pens began feeding faster and more birds resumed feeding than from low-complexity pens following playback of an alarm call, suggesting that birds housed in the complex environment were less anxious. Furthermore, birds housed in high-density or high-complexity pens had shorter tonic immobility durations on day 12 compared to day 26 in Experiment 1. In Experiment 2, birds from high-density pens had shorter tonic immobility durations than birds housed in low-density pens, which is contrary to expectations. Our results suggest that birds at 3 weeks of age were less fearful under high stocking density conditions than low density conditions. In addition, results indicated that the complex environment improved welfare of broilers through reduced anxiety.

Optimism and pasture access in dairy cows.

Allowing dairy cattle to access pasture can promote natural behaviour and improve their health. However, the psychological benefits are poorly understood. We compared a cognitive indicator of emotion in cattle either with or without pasture access. In a crossover experiment, 29 Holstein-Friesian dairy cows had 18 days of overnight pasture access and 18 days of full-time indoor housing. To assess emotional wellbeing, we tested cows on a spatial judgement bias task. Subjects learnt to approach a rewarded bucket location, but not approach another, unrewarded bucket location. We then presented cows with three "probe" buckets intermediate between the trained locations. Approaching the probes reflected an expectation of reward under ambiguity-an "optimistic" judgement bias, suggesting positive emotional states. We analysed the data using linear mixed-effects models. There were no treatment differences in latency to approach the probe buckets, but cows approached the known rewarded bucket slower when they had pasture access than when they were indoors full-time. Our results indicate that, compared to cattle housed indoors, cattle with pasture access display less anticipatory behaviour towards a known reward. This reduced reward anticipation suggests that pasture is a more rewarding environment, which may induce more positive emotional states than full-time housing.

Emotion in animal contests.

Emotions encompass cognitive and behavioural responses to reward and punishment. Using contests as a case-study, we propose that short-term emotions underpin animals' assessments, decision-making and behaviour. Equating contest assessments to emotional 'appraisals', we describe how contestants appraise more than resource value and outcome probability. These appraisals elicit the cognition, drive and neurophysiology that governs aggressive behaviour. We discuss how recent contest outcomes induce long-term moods, which impact subsequent contest behaviour. Finally, we distinguish between integral (objectively relevant) and incidental (objectively irrelevant) emotions and moods (affective states). Unlike existing ecological models, our approach predicts that incidental events influence contest dynamics, and that contests become incidental influences themselves, potentially causing maladaptive decision-making. As affective states cross contexts, a more holistic ethology (incorporating emotions and moods) would illuminate animal cognition and behaviour.

Microplastics disrupt hermit crab shell selection.

Microplastics (plastics < 5 mm) are a potential threat to marine biodiversity. However, the effects of microplastic pollution on animal behaviour and cognition are poorly understood. We used shell selection in common European hermit crabs (Pagurus bernhardus) as a model to test whether microplastic exposure impacts the essential survival behaviours of contacting, investigating and entering an optimal shell. We kept 64 female hermit crabs in tanks containing either polyethylene spheres (n = 35) or no plastic (n = 29) for 5 days. We then transferred subjects into suboptimal shells and placed them in an observation tank with an optimal alternative shell. Plastic-exposed hermit crabs showed impaired shell selection: they were less likely than controls to contact optimal shells or enter them. They also took longer to contact and enter the optimal shell. Plastic exposure did not affect time spent investigating the optimal shell. These results indicate that microplastics impair cognition (information-gathering and processing), disrupting an essential survival behaviour in hermit crabs.

Pasture Access Affects Behavioral Indicators of Wellbeing in Dairy Cows.

Dairy cows are increasingly housed indoors, either year-round or for long stretches over the winter and around parturition. This may create health and welfare issues. In cattle, lying and walking are highly motivated, and herds synchronize lying behavior when they have comfortable surfaces and little competition for space. Lying and walking activity can, therefore, indicate good welfare. Using a repeated measures crossover design, we gave 29 Holstein-Friesian dairy cows 18 days of overnight pasture access (PAS treatment) and 18 days of indoor housing (PEN treatment). Accelerometers recorded their lying and locomotory behavior. We measured behavioral synchrony with Fleiss' Kappa and analyzed the accelerometry data using linear mixed models. Compared to the PEN treatment, the PAS treatment had longer overnight lying durations (χ21 = 27.51, p < 0.001), fewer lying bouts (χ21 = 22.53, p < 0.001), longer lying bouts (χ21 = 25.53, p < 0.001), and fewer transitions up or down (χ21 = 16.83, p < 0.001). Herd lying behavior was also more synchronous at pasture (χ21 = 230.25, p < 0.001). In addition, nightly step counts were higher in the PAS treatment than the PEN treatment (χ21 = 2946.31, p < 0.001). These results suggest pasture access improves dairy cow welfare by increasing comfort, reducing competition and boredom, and facilitating motivated behavior.

Affect-Driven Attention Biases as Animal Welfare Indicators: Review and Methods.

Attention bias describes the differential allocation of attention towards one stimulus compared to others. In humans, this bias can be mediated by the observer's affective state and is implicated in the onset and maintenance of affective disorders such as anxiety. Affect-driven attention biases (ADABs) have also been identified in a few other species. Here, we review the literature on ADABs in animals and discuss their utility as welfare indicators. Despite a limited research effort, several studies have found that negative affective states modulate attention to negative (i.e., threatening) cues. ADABs influenced by positive-valence states have also been documented in animals. We discuss methods for measuring ADAB and conclude that looking time, dot-probe, and emotional spatial cueing paradigms are particularly promising. Research is needed to test them with a wider range of species, investigate attentional scope as an indicator of affect, and explore the possible causative role of attention biases in determining animal wellbeing. Finally, we argue that ADABs might not be best-utilized as indicators of general valence, but instead to reveal specific emotions, motivations, aversions, and preferences. Paying attention to the human literature could facilitate these advances.