Efficacy of a standardized training on horse welfare indicators: a preliminary study

Harmonized data collection is essential to obtain a reliable picture of equine welfare conditions. Effective education on how to assess and score welfare indicators plays a critical role in terms of inter-observer reliability. The Horse Grimace Scale (HGS), a facial-expression-based pain coding system, is able to identify a range of acute pain conditions in horses. This study aimed at evaluating the efficacy of a standardized training on HGS inter-observer reliability.Students in Veterinary Medicine from the University of Milan (N=46) and the University of Teramo (N=31) were recruited. Prior to any training, students were asked to score 10 pictures of horse faces using the six Facial Action Units (FAUs) of the HGS: Stiffly backwards ears, Orbital tightening, Tension above the eye area, Prominent strained chewing muscles, Mouth strained, Strained nostrils. Then, a 30-min training session was provided, including detailed descriptions and example pictures of each FAU, as well as a discussion of five pictures previously scored by an experienced assessor. After training, students scored other 10 pictures. To determine the inter-observer reliability pre and post-training, Intra-class Correlation Coefficient (ICC) was used.Students’ reliability was good even before training (ICC=0,986 for the overall HGS score), with Tension above the eye area, and Strained nostrils appearing more challenging to be scored reliably. Reliability improved after the 30 min training for the overall HGS score (ICC=0,992) and for each FAU (see table 1). According to Cicchetti (1994), an ICC score between 0.75 and 1.00 can be considered excellent.Our results suggest that the HGS scoring system is easy to apply even without any training; however, the training method applied proved useful to improve the reliability of HGS scores

Does poor stunning affect the assessment of animal-based measures consciousness in lambs at slaughter?

Oral session 2[EN] According to EU legislation on the killing of animals (EC n. 1099/2009), to ensure that animals do not regain consciousness before slaughter, the efficiency of stunning method must be evaluated through animal-based measures (ABMs)

Use of Qualitative Behaviour Assessment as an Indicator of Welfare in Donkeys

One of the objectives of the Animal Welfare Indicators project was to develop animal-based indicators to assess donkey welfare, including their emotional state. This study aimed to develop a fixed rating scale of Qualitative Behaviour Assessment (QBA) for donkeys, to evaluate the inter-observer reliability when applied on-farm, and to assess whether the QBA outcomes correlate to other welfare measures. A fixed list of 16 descriptors was designed on the basis of a consultation in a focus group. The fixed list was then used by four trained observers to score nine 2 min videos of groups of donkeys owned by six farms and on-farm to score 11 donkey facilities representative of the most common type of donkey facilities in Western Europe. On each farm one experienced assessor collected different welfare measures on all the adult donkeys. The QBA scores and welfare measures were analysed using Principal Component Analysis (PCA, correlation matrix, no rotation). Kendall’s W and ANOVA were used to assess inter-observer reliability. PCA revealed three main components explaining 79% of total variation between them. PC1ranged from at ease/relaxed to aggressive/uncomfortable, suggesting that this Component is important in the description of the valence of donkeys’ affective states. PC2 was more related to the level of arousal of donkeys, ranging from apathetic to distressed/responsive. The four assessors showed a good level of agreement on the first two dimensions of the PCA (Kendall’s W varying from 0.61 to 0.90), and there was no significant effect of observer on donkey QBA scores (ANOVA p \u3e 0.05), both for the videos and on-farm. PCA of all measures together showed positive QBA descriptors on PC1 (relaxed, at ease, happy, friendly) to be associated with positive human–donkey interaction indicators (absence of tail tuck, no avoidance, and positive reaction to an assessor walking down the side of the donkey). Our findings suggest that QBA is a suitable tool to identify the emotional state of donkeys on-farm. A fixed list of descriptors can be used consistently by different trained assessors as a valid addition to a number of animal welfare assessment indicators

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.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

Evaluation of Sheep Anticipatory Response to a Food Reward by Means of Functional Near-Infrared Spectroscopy

Anticipatory behaviour to an oncoming food reward can be triggered via classical conditioning, implies the activation of neural networks, and may serve to study the emotional state of animals. The aim of this study was to investigate how the anticipatory response to a food reward affects the cerebral cortex activity in sheep. Eight ewes from the same flock were trained to associate a neutral auditory stimulus (water bubble) to the presence of a food reward (maize grains). Once conditioned, sheep were trained to wait 15 s behind a gate before accessing a bucket with food (anticipation phase). For 6 days, sheep were submitted to two sessions of six consecutive trials each. Behavioural reaction was filmed and changes in cortical oxy- and deoxy-hemoglobin concentration ([ΔO2Hb] and [ΔHHb] respectively) following neuronal activation were recorded by functional near infrared spectroscopy (fNIRS). Compared to baseline, during the anticipation phase sheep increased their active behaviour, kept the head oriented to the gate (Wilcoxon’s signed rank test; p ≤ 0.001), and showed more asymmetric ear posture (Wilcoxon’s signed rank test; p ≤ 0.01), most likely reflecting a learnt association and an increased arousal. Results of trial-averaged [ΔO2Hb] and [ΔHHb] within individual sheep showed in almost every sheep a cortical activation during the anticipation phase (Student T-test; p ≤ 0.05). The sheep showed a greater response of the right hemisphere compared to the left hemisphere, possibly indicating a negative affective state, such as frustration. Behavioural and cortical changes observed during anticipation of a food reward reflect a learnt association and an increased arousal, but no clear emotional valence of the sheep subjective experience. Future work should take into consideration possible factors affecting the accurateness of measures, such as probe’s location and scalp vascularization

A Systematic Review on Commercially Available and Validated Sensor Technologies for Welfare Assessment of Dairy Cattle

In order to base welfare assessment of dairy cattle on real-time measurement, integration of valid and reliable precision livestock farming (PLF) technologies is needed. The aim of this study was to provide a systematic overview of externally validated and commercially available PLF technologies, which could be used for sensor-based welfare assessment in dairy cattle. Following Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, a systematic literature review was conducted to identify externally validated sensor technologies. Out of 1,111 publications initially extracted from databases, only 42 studies describing 30 tools (including prototypes) met requirements for external validation. Moreover, through market search, 129 different retailed technologies with application for animal-based welfare assessment were identified. In total, only 18 currently retailed sensors have been externally validated (14%). The highest validation rate was found for systems based on accelerometers (30% of tools available on the market have validation records), while the lower rates were obtained for cameras (10%), load cells (8%), miscellaneous milk sensors (8%), and boluses (7%). Validated traits concerned animal activity, feeding and drinking behavior, physical condition, and health of animals. The majority of tools were validated on adult cows. Non-active behavior (lying and standing) and rumination were the most often validated for the high performance. Regarding active behavior (e.g., walking), lower performance of tools was reported. Also, tools used for physical condition (e.g., body condition scoring) and health evaluation (e.g., mastitis detection) were classified in lower performance group. The precision and accuracy of feeding and drinking assessment varied depending on measured trait and used sensor. Regarding relevance for animal-based welfare assessment, several validated technologies had application for good health (e.g., milk quality sensors) and good feeding (e.g., load cells, accelerometers). Accelerometers-based systems have also practical relevance to assess good housing. However, currently available PLF technologies have low potential to assess appropriate behavior of dairy cows. To increase actors' trust toward the PLF technology and prompt sensor-based welfare assessment, validation studies, especially in commercial herds, are needed. Future research should concentrate on developing and validating PLF technologies dedicated to the assessment of appropriate behavior and tools dedicated to monitoring the health and welfare in calves and heifers

Bovine pestivirus is a new alternative virus for multiple myeloma oncolytic virotherapy

The oncolytic viruses have shown promising results for the treatment of multiple myeloma. However, the use of human viruses is limited by the patients' antiviral immune response. In this study, we investigated an alternative oncolytic strategy using non-human pathogen viruses as the bovine viral diarrhea virus (BVDV) that were able to interact with CD46

Non-invasive estimation of in vivo optical properties and hemodynamic parameters of domestic animals: a preliminary study on horses, dogs, and sheep

Biosensors applied in veterinary medicine serve as a noninvasive method to determine the health status of animals and, indirectly, their level of welfare. Near infrared spectroscopy (NIRS) has been suggested as a technology with this application. This study presents preliminary in vivo time domain NIRS measurements of optical properties (absorption coefficient, reduced scattering coefficient, and differential pathlength factor) and hemodynamic parameters (concentration of oxygenated hemoglobin, deoxygenated hemoglobin, total hemoglobin, and tissue oxygen saturation) of tissue domestic animals, specifically of skeletal muscle (4 dogs and 6 horses) and head (4 dogs and 19 sheep). The results suggest that TD NIRS in vivo measurements on domestic animals are feasible, and reveal significant variations in the optical and hemodynamic properties among tissue types and species. In horses the different optical and hemodynamic properties of the measured muscles can be attributed to the presence of a thicker adipose layer over the muscle in the Longissimus Dorsi and in the Gluteus Superficialis as compared to the Triceps Brachii. In dogs the absorption coefficient is higher in the head (temporalis musculature) than in skeletal muscles. The smaller absorption coefficient for the head of the sheep as compared to the head of dogs may suggest that in sheep we are indeed reaching the brain cortex while in dog light penetration can be hindered by the strongly absorbing muscle covering the cranium

The Feasibility of Animal-Based Indicators of Consciousness and Unconsciousness for Stunning in Sheep: A Systematic Review

Background: According to EU legislation, animal-based indicators (ABMs) are used to evaluate the efficacy of stunning methods to ensure that animals do not regain consciousness. EFSA has provided a list of ABMs for electrical and mechanical stunning in sheep; however, there is still a lack of information on their feasibility. We aimed to identify and evaluate the feasibility constraints of ABMs commonly applied in slaughterhouses to assess proper stunning in sheep. Method: For this systematic review, we searched the Scopus and Web of Science databases from 2000 to 8 August 2022, including full peer-reviewed papers written in English on the welfare of sheep at the stunning and restraint phases. We excluded studies using a gas stunning method or without prior stunning, as well as manuscripts in which indicators were applied after sticking. Results: Of 1289 records identified, only 8 papers were eligible for the critical evaluation of physical aspects that affect the feasibility of ABMs. These aspects were defined as a given definition of the feasibility of ABMs, and information was summarized and critically evaluated. The results highlighted a lack of information on the feasibility of ABMs which should be considered in the various conditions of commercial slaughterhouses

Development and Validation of a Test for the Classification of Horses as Broken or Unbroken

Regulation EC 1/2005 has stricter rules for transportation of unbroken (untamed) vs. broken (tamed) horses, but does not provide adequate tools for their identification. This study aimed to develop and validate such a tool. A behavioural test (Broken/Unbroken Test (BUT)) based on approaching, haltering, and leading was applied to 100 horses. Physiological and additional behavioural data were also collected, and the horses’ status (broken/unbroken) was assessed by the expert who administered the BUT. Each horse’s behaviour during the BUT was scored by four trained observers blinded to the horse’s history. The BUT score showed excellent inter-observer, intra-observer, and test–retest reliability (all intraclass correlation coefficients (ICCs) > 0.75). It was also negatively associated with respiratory rate, avoidance distance, and time needed to approach, halter, and lead the horse (p < 0.05 for all). The optimal BUT score cut-off for discrimination between broken and unbroken horses (gold standard: expert judgment) showed 97.8% sensitivity and 97.3% specificity. There was almost perfect agreement between BUT-based and expert classification of horses (ICC = 0.940). These findings confirm the BUT’s construct and criterion validity. The BUT could provide officials with a feasible, reliable, and valid tool to identify a horse’s broken/unbroken status and, consequently, direct stakeholders towards correct transport procedures