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

Recognised-by-law versus other identification systems in pigs: piglets discomfort evaluation and performance testing

The aim of the study was to evaluate the performance of recognised by Italian law (tattoos) and other (ear tags and injectable transponders) identification systems and to investigate if they caused different levels of short-term discomfort in piglets. Ninety-six ten-day-old piglets – equally assigned to four experimental groups – were identified with electronic ear tags (EET), transponders injected in the auricle base (TAB), in intraperitoneal position (TIP), or ear tattoo (T). Losses, breakages and readability of the identification devices were recorded to evaluate their efficiency. Forty-eight piglets were observed continuously for 5 min after identification. Behavioural indicators of pain, discomfort or ease were recorded in terms of duration of occurrence or frequency. Non-parametric analyses of variance were used to compare time required for the application of the devices and behavioural reactions of piglets. No healing problems or breakages of the electronic devices were observed during the trial. The readability after nine months was 93.8% for EET and TIP, 86.7% for TAB and 0% for T. The time required for the application differed significantly (P<0.001) among the four identification devices. EET and TIP showed better results than T considering losses, breakages, readability and time required for application. Behaviours most frequently recorded were head shaking and ear scratching, observed more frequently in piglets after the application of EET (P<0.001). The application of these identification devices caused a mild discomfort in piglets; further studies should investigate in detail the intensity and duration of reactions to the identification procedure

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)

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\u2019 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\u2019 and journeys\u2019 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\u2019 recall bias. Nevertheless, the results are in concordance with the literature, confirming that horse transport is a risk for the horse\u2019s and handler\u2019s 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

Conflict Behavior in Show Jumping Horses: A Field Study

peer-reviewedThe study objective was to determine if there was a relationship between behavioral and physiological stress measures in sport horses and their performance. Nineteen horses competed in show jumping events (6 housed at the center and 13 transported), while 5 horses at home training served as controls. The competition horses were assigned to “light” (obstacles ≤100 cm) and “difficult” class (obstacles >100 cm). The conflict behaviors (CBs/min) in two rounds were calculated. Total faults were classified as “less faults” (≤one fault) or “more faults” (>one fault). Salivary cortisol concentration (SCC) before the first round (SCC-SP1), 20 minutes (SCC-SP2), and 60 minutes after the second round (SCC-SP3) was measured. The increase (SCC-in) and decrease (SCC-dec) in SCC were calculated. No effect of competition was found. Horses that waited longer for the second round had greater CB (P < .05). Conflict behavior was more frequent in horses from the “more faults” (P = .05) and “difficult” (a tendency; P = .06) classes. No correlation of CB with SCC was found. SCC-SP2 was greater in “more faults” (P < .01) and “transported” (P < .01) horses. Competition increased the SCC (P < .05), whereas SCC-SP2 was greater in less successful horses (P < .05). Transported horses and horses with more faults had the greatest SCC-SP2 and SCC-dec (P < .05). Our results suggest that horses which presented stress response were also less successful in competition. The adoption of effective methods to reduce transport and competition stress could enhance welfare and performance of sport horses during competition

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

Identification of Altered miRNAs in Cerumen of Dogs Affected by Otitis Externa

Otitis externa is one of the most common diseases in dogs. It is associated with bacteria and yeast, which are regarded as secondary causes. Cerumen is a biological substance playing an important role in the protection of ear skin. The involvement of cerumen in immune defense is poorly understood. MicroRNAs can modulate the host immune response and can provide promising biomarkers for several inflammatory and infectious disorder diagnosis. The aims of this study were to profile the cerumen miRNA signature associated with otitis externa in dogs, integrate miRNAs to their target genes related to immune functions, and investigate their potential use as biomarkers. Cerumen was collected from healthy and otitis affected dogs and the expression of miRNAs was profiled by Next Generation Sequencing; the validation of the altered miRNAs was performed using RT-qPCR. The potential ability of miRNAs to modulate immune-related genes was investigated using bioinformatics tools. The results pointed out that 32 miRNAs, of which 14 were up- and 18 down-regulated, were differentially expressed in healthy vs. otitis-affected dogs. These results were verified by RT-qPCR. To assess the diagnostic value of miRNAs, ROC analysis was carried out, highlighting that 4 miRNAs are potential biomarkers to discriminate otitis-affected dogs. Bioinformatics showed that cerumen miRNAs may be involved in the modulation of host immune response. In conclusion, we have demonstrated for the first time that miRNAs can be efficiently extracted and quantified from cerumen, that their profile changes between healthy and otitis affected dogs, and that they may serve as potential biomarkers. Further studies are necessary to confirm their diagnostic value and to investigate their interaction with immune-related genes

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