Qualitative Behavioural Assessment as a welfare indicator for farmed Atlantic salmon (Salmo salar) in response to a stressful challenge.

Animal welfare assessments have struggled to investigate the emotional states of animals while focusing solely on available empirical evidence. Qualitative Behavioural Assessment (QBA) may provide insights into an animal's subjective experiences without compromising scientific rigor. Rather than assessing explicit, physical behaviours (i.e., what animals are doing, such as swimming or feeding), QBA describes and quantifies the overall expressive manner in which animals execute those behaviours (i.e., how relaxed or agitated they appear). While QBA has been successfully applied to scientific welfare assessments in a variety of species, its application within aquaculture remains largely unexplored. This study aimed to assess QBA's effectiveness in capturing changes in the emotional behaviour of Atlantic salmon following exposure to a stressful challenge. Nine tanks of juvenile Atlantic salmon were video-recorded every morning for 15 min over a 7-day period, in the middle of which a stressful challenge (intrusive sampling) was conducted on the salmon. The resultant 1-min, 63 video clips were then semi-randomised to avoid predictability and treatment bias for QBA scorers. Twelve salmon-industry professionals generated a list of 16 qualitative descriptors (e.g., relaxed, agitated, stressed) after viewing unrelated video-recordings depicting varying expressive characteristics of salmon in different contexts. A different group of 5 observers, with varied experience of salmon farming, subsequently scored the 16 descriptors for each clip using a Visual Analogue Scale (VAS). Principal Components Analysis (correlation matrix, no rotation) was used to identify perceived patterns of expressive characteristics across the video-clips, which revealed 4 dimensions explaining 74.5% of the variation between clips. PC1, ranging from 'relaxed/content/positive active' to 'unsettled/stressed/spooked/skittish' explained the highest percentage of variation (37%). QBA scores for video-clips on PC1, PC2, and PC4 achieved good inter- and intra-observer reliability. Linear Mixed Effects Models, controlled for observer variation in PC1 scores, showed a significant difference between PC1 scores before and after sampling (p = 0.03), with salmon being perceived as more stressed afterwards. PC1 scores also correlated positively with darting behaviours (r = 0.42, p < 0.001). These results are the first to report QBA's sensitivity to changes in expressive characteristics of salmon following a putatively stressful challenge, demonstrating QBA's potential as a welfare indicator within aquaculture.

Digital Livestock Technologies as boundary objects: Investigating impacts on farm management and animal welfare.

Digital Livestock Technologies (DLTs) can assist farmer decision-making and promise benefits to animal health and welfare. However, the extent to which they can help improve animal welfare is unclear. This study explores how DLTs may impact farm management and animal welfare by promoting learning, using the concept of boundary objects. Boundary objects may be interpreted differently by different social worlds but are robust enough to share a common identity across them. They facilitate communication around a common issue, allowing stakeholders to collaborate and co-learn. The type of learning generated may impact management and welfare differently. For example, it may help improve existing strategies (single-loop learning), or initiate reflection on how these strategies were framed initially (double-loop learning). This study focuses on two case studies, during which two DLTs were developed and tested on farms. In-depth, semi-structured interviews were conducted with stakeholders involved in the case studies (n = 31), and the results of a separate survey were used to complement our findings. Findings support the important potential of DLTs to help enhance animal welfare, although the impacts vary between technologies. In both case studies, DLTs facilitated discussions between stakeholders, and whilst both promoted improved management strategies, one also promoted deeper reflection on the importance of animal emotional well-being and on providing opportunities for positive animal welfare. If DLTs are to make significant improvements to animal welfare, greater priority should be given to DLTs that promote a greater understanding of the dimensions of animal welfare and a reframing of values and beliefs with respect to the importance of animals' well-being.

Once bitten, twice shy: Aggressive and defeated pigs begin agonistic encounters with more negative emotions.

Aggression between unfamiliar commercial pigs is common and likely invokes strong emotions in contestants. Furthermore, contest outcomes affect subsequent aggressive behaviour, suggesting a potential lasting influence on affective state. Here we used a combination of qualitative and quantitative methods to assess the emotional expression of pigs in agonistic encounters. We investigated how recent victory or defeat influences emotions expressed in a subsequent contest, and the role of aggressiveness as a personality trait in emotional expression. We observed the pre-escalation contest behaviour (second contest; age 13 wks) in animals of different aggressiveness (categorised using two resident intruder tests as Agg+ or Agg-, age 9 wks), which had recently won or lost a contest (first contest; 10 wks). We measured gaze direction and ear position. Observers watched video clips of the initial 30 s of the second contest and evaluated the emotional expression of 57 pigs (25 contest 1 winners, 32 contest 1 losers) using qualitative behavioural assessment (QBA) with a fixed list of 20 descriptive terms. QBA identified three principal components (PCs), accounting for 68% of the variation: PC1 (agitated/tense to relaxed/content), PC2 (fearful/aimless to confident/enjoying) and PC3 (listless/ indifferent). Agg- pigs and males showed a more positive emotionality (PC2). PC1 and PC3 were unaffected by first contest outcome and aggressiveness. Agg+ pigs were more likely to hold their ears back (X2 =7.8, p = 0.005) during the early contest period. Differences in attention were detected in the contest outcome × aggressiveness interaction (χ24.3, p = 0.04), whereby approaching the opponent was influenced by winning and losing in the Agg- pigs only. QBA and gaze behaviour reveal differences in emotional valence between pigs of different aggressiveness: less aggressive pigs may be more susceptible to the emotional impact of victory and defeat but overall, more aggressive pigs express more negative emotionality at the start of agonistic encounters.

Qualitative Behavioral Assessment in Juvenile Farmed Atlantic Salmon (Salmo salar): Potential for On-Farm Welfare Assessment.

There is a growing scientific and legislative consensus that fish are sentient, and therefore have the capacity to experience pain and suffering. The assessment of the welfare of farmed fish is challenging due to the aquatic environment and the number of animals housed together. However, with increasing global production and intensification of aquaculture comes greater impetus for developing effective tools which are suitable for the aquatic environment to assess the emotional experience and welfare of farmed fish. This study therefore aimed to investigate the use of Qualitative Behavioral Assessment (QBA), originally developed for terrestrial farmed animals, in farmed salmon and evaluate its potential for use as a welfare monitoring tool. QBA is a "whole animal" approach based on the description and quantification of the expressive qualities of an animal's dynamic style of behaving, using descriptors such as relaxed, agitated, lethargic, or confident. A list of 20 qualitative descriptors was generated by fish farmers after viewing video-footage showing behavior expressions representative of the full repertoire of salmon in this context. A separate, non-experienced group of 10 observers subsequently watched 25 video clips of farmed salmon, and scored the 20 descriptors for each clip using a Visual Analog Scale (VAS). To assess intra-observer reliability each observer viewed the same 25 video clips twice, in two sessions 10 days apart, with the second clip set presented in a different order. The observers were unaware that the two sets of video clips were identical. Data were analyzed using Principal Component (PC) Analysis (correlation matrix, no rotation), revealing four dimensions that together explained 79% of the variation between video clips, with PC1 (Tense/anxious/skittish-Calm/mellow/relaxed) explaining the greatest percentage of variation (56%). PC1 was the only dimension to show acceptable inter- and intra-observer reliability, and mean PC1 scores correlated significantly to durations of slow and erratic physical movements measured for the same 25 video clips. Further refinements to the methodology may be necessary, but this study is the first to provide evidence for the potential of Qualitative Behavioral Assessment to serve as a time-efficient welfare assessment tool for juvenile salmon under farmed conditions.

High-starch diets alter equine faecal microbiota and increase behavioural reactivity.

Gut microbiota have been associated with health, disease and behaviour in several species and are an important link in gut-brain axis communication. Diet plays a key role in affecting the composition of gut microbiota. In horses, high-starch diets alter the hindgut microbiota. High-starch diets are also associated with increased behavioural reactivity in horses. These changes in microbiota and behaviour may be associated. This study compares the faecal microbiota and behaviour of 10 naïve ponies. A cross-over design was used with experimental groups fed high-starch (HS) or high-fibre (HF) diets. Results showed that ponies were more reactive and less settled when being fed the HS diet compared to the HF diet. Irrespective of diet, the bacterial profile was dominated by two main phyla, Firmicutes, closely followed by Bacteroidetes. However, at lower taxonomic levels multivariate analysis of 16S rRNA gene sequencing data showed diet affected faecal microbial community structure. The abundance of 85 OTUs differed significantly related to diet. Correlative relationships exist between dietary induced alterations to faecal microbiota and behaviour. Results demonstrate a clear link between diet, faecal microbial community composition and behaviour. Dietary induced alterations to gut microbiota play a role in affecting the behaviour of the host.

Using Qualitative Behaviour Assessment to Investigate Human-Animal Relationships in Zoo-Housed Giraffes (Giraffa camelopardalis).

Human-Animal Relationships (HAR) in zoos develop from repeated interactions between animals and their caretakers. HAR have been shown to affect health and welfare in farm animals, but limited zoo-based studies exist. This study investigates the association between the qualitative behaviour assessment (QBA) of emotional expression in giraffes and keeper action score in four types of keeper-animal interaction (KAI). Three giraffes generating 38 clips. QBA, using a free-choice profiling methodology, was applied instructing 18 observers to assess giraffe expressions shown in these clips. QBA scores were analysed using Generalized Procrustes Analysis. Keeper actions during each KAI event were rated by an independent marker, resulting in cumulative scores for keeper action quality. The association between QBA and the keeper action was analyzed using Spearman's rank correlations. Two main QBA dimensions were identified explaining 59% of the variation between clips. There were significant effects of giraffe and KAI type on QBA dimension 2 (inquisitive/impatient-calm/distracted), and significant positive associations between keeper action quality rating and QBA dimensions 1 and 2, indicating that positive keeper actions resulted in calm and confident giraffes with a willingness to interact. This is the first successful application of QBA for empirically addressing HARs in zoos, however given the small sample size of giraffes in this study, it can be regarded as a pilot study only, and further research is needed to validate the use of QBA in this context.

The Assessment of Landscape Expressivity: A Free Choice Profiling Approach.

In this paper we explore a relational understanding of landscape qualities. We asked three independent groups of human observers to assess the expressive qualities of a range of landscapes in the UK and in Spain, either by means of personal visits or from a projected digital image. We employed a Free Choice Profiling (FCP) methodology, in which observers generated their own descriptive terminologies and then used these to quantify perceived landscape qualities on visual analogue scales. Data were analysed using Generalised Procrustes Analysis, a multivariate statistical technique that does not rely on fixed variables to identify underlying dimensions of assessment. The three observer groups each showed significant agreement, and generated two main consensus dimensions that suggested landscape 'health' and 'development in time' as common perceived themes of landscape expressivity. We critically discuss these outcomes in context of the landscape assessment literature, and suggest ways forward for further development and research.

Evaluation of Animal-Based Indicators to Be Used in a Welfare Assessment Protocol for Sheep.

Sheep are managed under a variety of different environments (continually outdoors, partially outdoors with seasonal or diurnal variation, continuously indoors) and for different purposes, which makes assessing welfare challenging. This diversity means that resource-based indicators are not particularly useful and, thus, a welfare assessment scheme for sheep, focusing on animal-based indicators, was developed. We focus specifically on ewes, as the most numerous group of sheep present on farm, although many of the indicators may also have relevance to adult male sheep. Using the Welfare Quality® framework of four Principles and 12 Criteria, we considered the validity, reliability, and feasibility of 46 putative animal-based indicators derived from the literature for these criteria. Where animal-based indicators were potentially unreliably or were not considered feasible, we also considered the resource-based indicators of access to water, stocking density, and floor slipperiness. With the exception of the criteria "Absence of prolonged thirst," we suggest at least one animal-based indicator for each welfare criterion. As a minimum, face validity was available for all indicators; however, for many, we found evidence of convergent validity and discriminant validity (e.g., lameness as measured by gait score, body condition score). The reliability of most of the physical and health measures has been tested in the field and found to be appropriate for use in welfare assessment. However, for the majority of the proposed behavioral indicators (lying synchrony, social withdrawal, postures associated with pain, vocalizations, stereotypy, vigilance, response to surprise, and human approach test), this still needs to be tested. In conclusion, the comprehensive assessment of sheep welfare through largely animal-based measures is supported by the literature through the use of indicators focusing on specific aspects of sheep biology. Further work is required for some indicators to ensure that measures are reliable when used in commercial settings.

Validating the use of qualitative behavioral assessment as a measure of the welfare of sheep during transport.

We tested the application of qualitative behavioral assessment (QBA) as a welfare assessment tool. Sheep were exposed to road transport treatments, and behavioral expressions were compared between experimental treatments and validated by correlation with physiological measures. We compared journeys differing in ventilation (closed vs. open-sided trailer), flooring (grip vs. nongrip flooring), and driving styles (stop-start vs. continuous driving). Blood samples were collected immediately before loading and after unloading; heart rate and core body temperatures were recorded continuously. Continuous video footage was edited to show individual sheep to observers for QBA using free-choice profiling (observers used their own descriptive terms). There was significant consensus in observers' scores for the sheep in each experiment (p < .001). Observers distinguished between sheep exposed to flooring (p = .014) or driving-style (p = .005) treatments, but not between ventilation treatments. QBA scores were compared (p < .05) with plasma leptin, glucose, and insulin-like growth factor-1 concentrations; white blood cell profiles; red blood cell counts; hematocrit; body temperatures; and heart rate variability. Observer assessments reflected treatment differences, and correlations between behavioral expression and physiological responses were found.

Qualitative Behavioural Assessment of emotionality in pigs.

Scientific assessment of affective states in animals is challenging but vital for animal welfare studies. One possible approach is Qualitative Behavioural Assessment (QBA), a 'whole animal' methodology which integrates information from multiple behavioural signals and styles of behavioural expression (body language) directly in terms of an animal's emotional expression. If QBA provides a valid measure of animals' emotional state it should distinguish between groups where emotional states have been manipulated. To test this hypothesis, QBA was applied to video-recordings of pigs, following treatment with either saline or the neuroleptic drug Azaperone, in either an open field or elevated plus-maze test. QBA analysis of these recordings was provided by 12 observers, blind to treatment, using a Free Choice Profiling (FCP) methodology. Generalised Procrustes Analysis was used to calculate a consensus profile, consisting of the main dimensions of expression. Dimension one was positively associated with terms such as 'Confident' and 'Curious' and negatively with 'Unsure' and 'Nervous'. Dimension two ranged from 'Agitated'/'Angry' to 'Calm'/'Relaxed'. In both tests, Azaperone pre-treatment was associated with a more positive emotionality (higher scores on dimension one reflecting a more confident/curious behavioural demeanour) than control pigs. No effect of drug treatment on dimension two was found. Relationships between qualitative descriptions of behaviour and quantitative behavioural measures, taken from the same recordings, were found. Overall, this work supports the use of QBA for the assessment of emotionality in animals.