Does housing system affect horse welfare? The AWIN welfare assessment protocol applied to horses kept in an outdoor group-housing system: The 'parcours'.

Outdoor group housing is generally reported as being beneficial to the welfare of horses compared to single boxes, being considered to show greater similarities with the living conditions of feral horses, allowing full expression of behaviours such as grazing, social interactions and free movement. However, concerns persist regarding the ability to maintain a good nutritional state and the possibility of acquiring injury. No data reporting a comprehensive assessment of welfare for horses in outdoor group-housing systems are currently available. The present study aimed at applying a scientifically valid welfare assessment protocol to group-housed outdoor horses in 'parcours', a particular management system used in the south of France. 'Parcours' are semi-natural areas, grazed by domestic herbivores located in lowland, mountain, or marsh. One hundred and seventy-one horses older than a year pertaining to six farms and kept on 'parcours' were evaluated by a trained veterinarian using a modified version of the second level Animal Welfare Indicators (AWIN) welfare assessment protocol for horses. No major welfare issues were detected. Horses in 'parcours' displayed few abnormal behaviours, they could move freely for most of the day and interact with conspecifics, maintaining a healthy state of nutrition and a good relationship with humans. The main welfare concerns were related to the presence of superficial integument alterations such as alopecia, difficulty in reaching quality controlled water sources and a lack of shelter. As the number of facilities involved in this study is relatively limited, further harmonised data collection should aim to enlarge the sample size and allow comparison with different outdoor group-housing conditions.

Application of QBA to Assess the Emotional State of Horses during the Loading Phase of Transport.

To identify feasible indicators to evaluate animals' emotional states as a parameter to assess animal welfare, the present study aimed at investigating the accuracy of free choice profiling (FCP) and fixed list (FL) approach of Qualitative Behaviour Assessment (QBA) in horses during the loading phase of transport. A total of 13 stakeholders were trained to score 2 different sets of videos of mixed breed horses loaded for road transport, using both FCP and FL, in 2 sessions. Generalized Procustes Analysis (GPA) consensus profile explained a higher percentage of variation (80.8%) than the mean of 1000 randomized profiles (41.2 ± 1.6%; p = 0.001) for the FCP method, showing an excellent inter-observer agreement. GPA identified two main factors, explaining 65.1% and 3.7% of the total variation. Factor 1 ranging from 'anxious/ to 'calm/relaxed', described the valence of the horses' emotional states. Factor 2, ranging from 'bright' to 'assessing/withdrawn', described the arousal. As for FL, Principal Component Analysis (PCA) first and second components (PC1 and PC2, respectively), explaining on average 59.8% and 12.6% of the data variability, had significant agreement between observers. PC1 ranges from relaxed/confident to anxious/frightened, while PC2 from alert/inquisitive to calm. Our study highlighted the need for the use of descriptors specifically selected, throughout a prior FCP process for the situation we want to evaluate to get a good QBA accuracy level.

The Impact of Noise Anxiety on Behavior and Welfare of Horses from UK and US Owner's Perspective.

Noise anxiety is an over-reaction to loud noises commonly detected among pets and can greatly impact on their welfare and on their management. When exposed to noisy events, horses can show intense escape attempts, which may cause severe accidents for the horse and the rider/handler. The aim of the present study was to investigate, through a web survey, UK and US owners' perception of noise anxiety severity in their horses, their management strategies and perceived efficacy. The questionnaire was shared via social networking and advertised as "What is your horse afraid of?". Over a total of 1836 questionnaires filled out; 409 owners reported that their horse has shown unusual behavior during a noise event. A two-step cluster analysis identified two groups: very anxious (VA) and slightly anxious (SA). VA horses were reported to have higher frequency of anxiety behaviors; higher frequency of signs of noise reactivity; and their anxiety did not improve with time. The most used management strategies consisted in providing hay throughout the night, turning in/out their horse or moving it to a paddock. A binomial logistic regression identified that horses that have reported injuries during noise events were more likely to be clustered as VA (OR = 0.24, 95% CI: 0.08-0.76); while providing hay throughout the night was more likely to be very effective management strategy in SA horses (OR = 0.41, 95% CI: 0.16-1.01). Our results confirmed that noise anxiety is a growing behavioral problem that can lead to important welfare concerns for horses. New management strategies, including the use of medicinal products, should be considered to reduce behavioral and physiological signs and help horses to cope with noisy events.

Effects of BEMER® physical vascular therapy in horses under training. A randomized, controlled double blind study.

Horses preparation for competition may cause psychological and physical stress. Physical vascular therapy BEMER® is reported to increase vasomotion and microcirculation, supporting body healing. This study aimed at assessing whether BEMER® physical vascular therapy in horses influences recovery rate of hematological and biochemical blood parameters within 1 h after moderate exercise and reduces stress measured by physiological and behavioral indicators. This prospective, randomized, double blinded, placebo-controlled crossover study included twelve warmblood horses (3 mares, 8 geldings, 1 stallion). Additionally to their daily work, horses were subjected to 15 min of exercise on a longe. Horses were randomly divided in two groups: A (n = 6), B (n = 6). Group A underwent first to BEMER® blanket for two weeks, then to Placebo blanket for two weeks. Group B did the opposite. Blood samples, thermographic infrared images, Heart Rate Variability and behavior were analyzed. ANOVA was used to investigate any treatment effect. After two weeks of treatment, although not statistically significant, hematocrit (%) measured immediately post exercise was lower in horses undergoing BEMER® treatment (48.30% ± 3.21) than both No blanket (51.15% ± 3.57) and Placebo blanket (49.58% ± 5.77). While wearing BEMER® blanket and after treatment, horses had a lower LF/HF ratio compared to other groups, although this difference was not statistically significant. These results possibly suggest an effect of BEMER® therapy on vagal activity and relaxation. Substantial progress in recovery after exercise was not confirmed, leading to the need for further investigation on the overall effect of BEMER® therapy.

Equine Transport-Related Problem Behaviors and Injuries: A Survey of Italian Horse Industry Members.

An online survey was conducted to determine associations between equine transport management and transport-related injuries and problem behaviors in Italy. The survey was composed of four sections: respondents' demographic information and background, transport management practices, journey details and vehicle design, and transport injuries experienced by the horse in the previous two-year period. Univariable and multivariable logistic regression with a binary outcome variable was performed to explore associations between variables (respondents' and journeys' details and transport practices) and equine transport-related problem behaviors (TRPBs) and injuries. TRPBs were also considered an explanatory variable for injuries. The survey generated 201 responses; only 148 were complete and analyzed. TRPBs were reported by 14.45% of the respondents and the odds of TRPBs was linked to the respondent gender (p = 0.034), the use of tranquilizers prior to transport (p = 0.002), the use of a whip for loading (p = 0.049), the lack of protection equipment (p = 0.050), and shavings (p = 0.025) on the vehicle floor. Horse injuries (11.49%) were reported by more respondents who did not check the brakes of their transport vehicle before traveling (p = 0.043), had vehicles with padding on the chest bar (p = 0.038), and for horses reported to display TRPBs (p = 0.001). Finally, 10 respondents reported they were injured during horse transport (10/140; 7.14%), 50% simultaneously with their horses. The study findings should be interpreted with caution due to small sample size bias and participants' recall bias. Nevertheless, the results are in concordance with the literature, confirming that horse transport is a risk for the horse's and handler's health and well-being. Further studies are needed to identify best management practices to educate equine industry members on how to minimize transport-related problems.

Towards an improved pain assessment in castrated horses using facial expressions (HGS) and circulating miRNAs.

BACKGROUND: Pain in horses is an emergent welfare concern, and its assessment represents a challenge for equine clinicians. This study aimed at improving pain assessment in horses through a convergent validation of existing tools: we investigated whether an effective analgesic treatment influences the horse grimace scale (HGS) and the concentration of specific circulating microRNAs (miRNAs). METHODS: Eleven stallions underwent routine surgical castration under general anaesthesia. They were divided into two analgesic treatment groups: castration with the administration of preoperative flunixin and castration with preoperative flunixin plus a local injection of mepivacaine into the spermatic cords. HGS and levels of seven circulating miRNAs were evaluated pre-, 8 and 20 hours post-procedure. RESULTS: Compared to pre-castration, HGS, miR-126-5p, miR-145 and miR-let7e increased significantly in horses receiving flunixin at 8 hours post-castration (Friedman test, p < 0.05). Both behavioural and molecular changes occurred in horses receiving flunixin only, confirming that the addition of local mepivacaine is an effective analgesic treatment. CONCLUSIONS: Combining the use of HGS and circulating miRNAs, particularly miR-145, could be meaningful to monitor acute pain conditions in horses. Our results further validate the HGS as a method to assess acute pain in horses and point out miR-145 as a promising biomarker to identify pain.

Use of Detomidine Oromucosal Gel for Alleviation of Acute Anxiety and Fear in Horses: A Pilot Study.

The aim of this randomized, double-blind, placebo-controlled, parallel group clinical field study was to evaluate the effect of detomidine oromucosal gel in alleviating anxiety and fear in horses. Sixteen horses with a history of acute anxiety and fear associated with firework-related noise entered the study. On New Year's Eve, eight horses were treated with 30 µg/kg detomidine gel and eight horses with placebo gel. When fireworks were present, 75% (6/8) of the detomidine-treated horses were scored by their owners as having a good or excellent treatment effect on anxiety and fear, while 50% (3/6) of horses receiving placebo were scored to have a good effect. Horses' behavior was video-recorded and assessed with a focal animal continuous method by a treatment-blind expert observer. Results showed that when fireworks were present, walking behavior decreased significantly (p < 0.05) after treatment with detomidine and that horses of the placebo group, overall, showed more restlessness, vocalization, and signs of colic (Wilcoxon matched-pairs test on the first PC, p = 0.007). This study indicates that detomidine oromucosal gel can be used to alleviate acute noise-related anxiety and fear in horses, but larger treatment groups are needed to confirm the results.

Does Thirty-Minute Standardised Training Improve the Inter-Observer Reliability of the Horse Grimace Scale (HGS)? A Case Study.

The Horse Grimace Scale (HGS) is a facial-expression-based pain coding system that enables a range of acute painful conditions in horses to be effectively identified. Using valid assessment methods to identify pain in horses is of a clear importance; however, the reliability of the assessment is highly dependent on the assessors' ability to use it. Training of new assessors plays a critical role in underpinning reliability. The aim of the study was to evaluate whether a 30-minute standardised training program on HGS is effective at improving the agreement between observers with no horse experience and when compared to an HGS expert. Two hundred and six undergraduate students with no horse experience were recruited. Prior to any training, observers were asked to score 10 pictures of horse faces using the six Facial Action Units (FAUs) of the HGS. Then, an HGS expert provided a 30-minute face-to-face training session, including detailed descriptions and example pictures of each FAU. After training, observers scored 10 different pictures. Cohen's k coefficient was used to determine inter-observer reliability between each observer and the expert; a paired-sample t-test was conducted to determine differences in agreement pre- and post-training. Pre-training, Cohen's k ranged from 0.20 for tension above the eye area to 0.68 for stiffly backwards ears. Post-training, the reliability for stiffly backwards ears and orbital tightening significantly increased, reaching Cohen's k values of 0.90 and 0.91 respectively (paired-sample t-test; p < 0.001). The results suggest that this 30-minute face-to-face training session was not sufficient to allow observers without horse experience to effectively apply HGS. However, this standardised training program could represent a starting point for a more comprehensive training program for those without horse experience in order to increase their reliably in applying HGS.

Habituation to Transport Helps Reducing Stress-Related Behavior in Donkeys During Loading.

Adopting proper animal management strategies, including training, might reduce to a substantial extent the adverse effects of transport-related stress in animals. The aim of this study was to evaluate the effect of habituation to transport on stress-related behaviors and physiological indicators during loading and unloading in donkeys. Fourteen donkeys were recruited and divided in two treatment groups: Habituation (H; M = 5, F = 2) and Control (C; M = 5, F = 2). H donkeys were gradually habituated to be transported, traveling together with their mothers and other adult donkeys well-accustomed to transport, while C donkeys had never been transported before. Loading and unloading phases were video recorded and behavior was analyzed. Saliva samples for cortisol concentration determination were collected at rest and after unloading. Latency time to load was significantly shorter for H donkeys than C donkeys (Mann-Whitney; p = 0.004). C donkeys also showed significantly more stress-related behaviors (Mann-Whitney; p = 0.026) and required a higher but not statistically significant number of human interventions to load. Cortisol concentration increased in both groups, but no differences were found between them (Mann-Whitney; p > 0.05). These results suggest that habituation to transport could mitigate stress during loading procedures in donkeys reducing loading time, frequency of stress-related behaviors and diminishing the need of human intervention.

May Salivary Chromogranin A Act as a Physiological Index of Stress in Transported Donkeys? A Pilot Study.

Road transport is known to be a stressful animal husbandry procedure as it induces the activation of two main physiological stress-related pathways: the hypothalamic-pituitary-adrenal cortex axis and the sympathetic-adrenal medulla axis. This preliminary study aimed to investigate whether salivary chromogranin A (CgA) concentration, known as a biomarker of the sympathetic activity system during psychological stress, may represent a novel physiological index of transportation-induced stress in donkeys. Nineteen Romagnolo donkeys, raised in groups on paddocks, were subject to two transportations, following the farm's routine procedures, for a mean duration of 64 min each on two consecutive days. Salivary samples were gently collected 15 min before and 15 min after each transportation. Salivary CgA was measured by a commercially available enzyme-linked immunosorbent assay test. Results showed that CgA salivary levels significantly decreased after both transportations. The physiological mechanisms underlying this result may be related to catestatin activity, a bioactive product of the proteolytic cleavage of CgA, that acts as an inhibitor of catecholamine release. This hypothesis requires further investigation, particularly considering the limited number of subjects involved in this preliminary study. The identification of a reliable and non-invasive stress-marker would represent a useful tool for improving farm animals' welfare in transport conditions.

Positive Reinforcement-Based Training for Self-Loading of Meat Horses Reduces Loading Time and Stress-Related Behavior.

The present work aimed to evaluate the efficacy of a self-loading training using positive reinforcement on stress-related behaviors shown by meat horses during loading procedures into a truck. Thirty-two meat horses (M = 18; F = 14; 6 month-old) were included in the study. All horses had limited interactions with the farmer and were not used to be restrained nor lead by halter. Horses were divided in two groups: Control Group (C; N = 14) and Training Group (T; N = 18). T horses were trained to self-load: in order to teach the horses to enter into the truck, a targeting training technique throughout a shaping process was applied. Training sessions were performed three times a week, from 9:30 a.m. to 1:30 p.m. and from 2:30 p.m. to 4:30 p.m., for 6 weeks; training was then repeated once a week to maintain the memory until the transport toward a slaughterhouse. The loading phase was video-recorded and loading time was directly recorded using a stopwatch. All horses were transported to the same slaughterhouse in 14 different days using the same truck. Behavior was subsequently analyzed with a focal animal continuous recording method. Loading time was shorter in T horses (mean ± SD = 44.44 ± 47.58 s) than in C horses (mean ± SD = 463.09 ± 918.19 s) (T-test; p = 0.019). T horses showed more forward locomotion toward the truck than C horses (T-test; p = 0.029). Our preliminary findings suggest that self-loading training may be useful to mitigate loading-related stress in meat horses, minimally socialized with humans.

Text Mining Analysis to Evaluate Stakeholders' Perception Regarding Welfare of Equines, Small Ruminants, and Turkeys.

Welfare of animals significantly depends on how stakeholders perceive their needs and behave in a way to favor production systems that promote better welfare outcomes. This study aimed at investigating stakeholders' perception of the welfare of equines, small ruminants, and turkeys using text mining analysis. A survey composed by open-ended questions referring to different aspects of animal welfare was carried out. Text mining analysis was performed. A total of 270 surveys were filled out (horses = 122, sheep = 81, goats = 36, turkeys = 18, donkeys = 13). The respondents (41% veterinarians) came from 32 different countries. To describe welfare requirements, the words "feeding" and "water" were the most frequently used in all the species, meaning that respondents considered the welfare principle "good feeding" as the most relevant. The word "environment" was considered particularly important for turkeys, as well as the word "dry", never mentioned for other species. Horses stakeholders also considered "exercise" and "proper training" important. Goat stakeholders' concerns are often expressed by the word "space", probably because goats are often intensively managed in industrialized countries. Although the sample was too small to be representative, text mining analysis seems to be a promising method to investigate stakeholders' perception of animal welfare, as it emphasizes their real perception, without the constraints deriving by close-ended questions.

Can grimace scales estimate the pain status in horses and mice? A statistical approach to identify a classifier.

Pain recognition is fundamental for safeguarding animal welfare. Facial expressions have been investigated in several species and grimace scales have been developed as pain assessment tool in many species including horses (HGS) and mice (MGS). This study is intended to progress the validation of grimace scales, by proposing a statistical approach to identify a classifier that can estimate the pain status of the animal based on Facial Action Units (FAUs) included in HGS and MGS. To achieve this aim, through a validity study, the relation between FAUs included in HGS and MGS and the real pain condition was investigated. A specific statistical approach (Cumulative Link Mixed Model, Inter-rater reliability, Multiple Correspondence Analysis, Linear Discriminant Analysis and Support Vector Machines) was applied to two datasets. Our results confirm the reliability of both scales and show that individual FAU scores of HGS and MGS are related to the pain state of the animal. Finally, we identified the optimal weights of the FAU scores that can be used to best classify animals in pain with an accuracy greater than 70%. For the first time, this study describes a statistical approach to develop a classifier, based on HGS and MGS, for estimating the pain status of animals. The classifier proposed is the starting point to develop a computer-based image analysis for the automatic recognition of pain in horses and mice.

Welfare Conditions of Donkeys in Europe: Initial Outcomes from On-Farm Assessment.

This paper is a baseline study to present the initial outcomes of data collected in a sample of EU donkey farms using the AWIN welfare assessment protocol for donkeys, comprehensive of 22 valid, reliable and feasible animal-based indicators. A total of 20 donkey facilities (N = 12 in Italy and N = 8 in United Kingdom) were visited and 278 donkeys of different breed, aged 2-45 years, were assessed. Three assessors underwent a common training period to learn how to perform and score all the indicators included in the protocol. Data was collected using digitalized systems and downloaded to a database. A descriptive statistic for each welfare indicator was calculated. The authors found recurrent issues: 25% of donkeys were moderately over weight; although most of the assessed animals had good quality hoof care, 15.16% of them presented some signs of neglect, such as overgrowth and/or incorrect trimming; 18.05% of donkeys showed an avoidance reaction to an approaching human in the avoidance distance test. The protocol has proven to be applicable in different management conditions and for donkeys of different attitude.

Using the Horse Grimace Scale (HGS) to Assess Pain Associated with Acute Laminitis in Horses (Equus caballus).

Acute laminitis is a common equine disease characterized by intense foot pain, both acutely and chronically. The Obel grading system is the most widely accepted method for describing the severity of laminitis by equine practitioners, however this method requires movement (walk and trot) of the horse, causing further intense pain. The recently developed Horse Grimace Scale (HGS), a facial-expression-based pain coding system, may offer a more effective means of assessing the pain associated with acute laminitis. The aims of this study were: to investigate whether HGS can be usefully applied to assess pain associated with acute laminitis in horses at rest, and to examine if scoring HGS using videos produced similar results as those obtained from still images. Ten horses, referred as acute laminitis cases with no prior treatment, were included in the study. Each horse was assessed using the Obel and HGS (from images and videos) scales: at the admission (before any treatment) and at seven days after the initial evaluation and treatment. The results of this study suggest that HGS is a potentially effective method to assess pain associated with acute laminitis in horses at rest, as horses showing high HGS scores also exhibited higher Obel scores and veterinarians classified them in a more severe painful state. Furthermore, the inter-observer reliability of the HGS total score was good for both still images and video evaluation. There was no significant difference in HGS total scores between the still images and videos, suggesting that there is a possibility of applying the HGS in clinical practice, by observing the horse for a short time. However, further validation studies are needed prior to applying the HGS in a clinical setting.

The development of the AWIN welfare assessment protocol for donkeys / O desenvolvimento do protocolo de avaliação de bem-estar AWIN (Animal Welfare Indicators) para jumentos

The donkey population has increased in the last 10 years, with an estimated 50 million donkeys currently worldwide. Donkey welfare, meanwhile, is an increasing global concern that receives close public scrutiny. However, multiple challenges are surrounding how donkey welfare is assessed and recorded. The Animal Welfare Indicators (AWIN) project is the first project, funded by the European Commission, intended to improve donkey welfare by developing a scientifically sound and practical on-farm welfare assessment protocol. The present study describes the procedure for the development of the AWIN welfare assessment protocol for donkeys: 1) selection of promising welfare indicators; 2) research to cover gaps in knowledge; 3) stakeholder consultation; 4) testing the prototype protocol on-farm. The proposed two-level strategy improved on-farm feasibility, while the AWIN donkey app enables the standardized collection of data with prompt results. Although limitations are linked with a relatively small reference population, the AWIN welfare assessment protocol represents the first scientific and standardized approach to evaluate donkey welfare on-farm.(AU)

Na última década, a população de jumentos vem aumentando; estima-se que existam aproximadamente 50 milhões de em todo o mundo. O bem-estar dos jumentos é uma preocupação global crescente, que recebe um escrutínio público próximo. No entanto, existem vários desafios em torno de como o bem-estar do jumento é avaliado e registrado. O projeto Indicadores de Bem-Estar Animal (AWIN) foi o primeiro projeto, financiado pela Comissão Europeia, destinado a melhorar o bem-estar dos jumentos, desenvolvendo um protocolo de avaliação do bem-estar cientificamente válido e prático na fazenda. O presente estudo descreve o procedimento para o desenvolvimento do protocolo de avaliação de bem-estar AWIN para jumentos: 1) seleção de indicadores promissores de bem-estar; 2) pesquisa para cobrir lacunas no conhecimento; 3) consulta às partes interessadas; 4) testando o protocolo do protótipo em fazendas. A estratégia proposta em dois níveis de avaliação melhorou a viabilidade na fazenda, além disso, o aplicativo AWIN donkey permite coletar dados de maneira padronizada e mostrar resultados rapidamente. Embora a limitação esteja ligada a uma população de referência relativamente pequena, o protocolo de avaliação de bem-estar do AWIN representa a primeira abordagem científica e padronizada para avaliar o bem-estar de jumentos em fazendas.(AU)