Shape and joint angle data for seven European horse breeds and their repeatability.

Conformation traits are important in the selection and distinction between horse breeds, but tend to be evaluated subjectively within a breed and cannot be compared between them. The horse shape space model, using a combination of 253 landmarks and semi-landmarks, provides objective information on the shape of a horse photographed from the side that can be compared between breeds. In this dataset, we are providing the full set of 253 landmarks for 1241 horses from seven breeds, including an R code file to extract joint angle information and transform the raw data into csv files for further analysis, such as breed comparisons, heritability or genome-wide association studies (single- or multibreed). The repeatability of the joint angles are also reported.

Technique description and outcome evaluation of Thoroughbred racehorses following soft palate thermocautery performed under standing sedation.

BACKGROUND: Palatal dysfunction (PD), which encompasses palatal instability (PI) and intermittent dorsal displacement of the soft palate, is the most common performance-limiting upper respiratory tract obstruction in young Thoroughbred racehorses. Soft palate thermocautery (SPT) performed under general anaesthesia is a routinely performed procedure for PD in some countries, but the procedure for and outcome of SPT performed under standing sedation has not been published. OBJECTIVES: (1) To describe a technique for SPT performed under standing sedation; and (2) to assess post-operative performance in horses compared to controls using the Racing Post rating (RPR), British Horseracing Authority official rating (OR), Performance Index and Earnings. STUDY DESIGN: Retrospective case series. METHODS: Medical records were reviewed for all horses that had SPT performed under standing sedation following topical and local infusion of lidocaine hydrochloride into the rostral soft palate, and that were identified to have PI by overground endoscopy. Two matched controls were identified for each case. The median RPR, OR, Performance Index and Earnings for the three pre-operative and three post-operative races were compared. RESULTS: No significant differences were identified between the SPT (n = 23) and Control groups (n = 46) for baseline characteristics or outcomes. MAIN LIMITATIONS: Retrospective study design, small sample size. CONCLUSIONS: No significant differences in racing performance were identified between horses that had SPT performed under standing sedation and controls.

Breaking Latent Infection: How ORF37/38-Deletion Mutants Offer New Hope against EHV-1 Neuropathogenicity.

Equid alphaherpesvirus 1 (EHV-1) has been linked to the emergence of neurological disorders, with the horse racing industry experiencing significant impacts from outbreaks of equine herpesvirus myeloencephalopathy (EHM). Building robust immune memory before pathogen exposure enables rapid recognition and elimination, preventing infection. This is crucial for effectively managing EHV-1. Removing neuropathogenic factors and immune evasion genes to develop live attenuated vaccines appears to be a successful strategy for EHV-1 vaccines. We created mutant viruses without ORF38 and ORF37/38 and validated their neuropathogenicity and immunogenicity in hamsters. The ∆ORF38 strain caused brain tissue damage at high doses, whereas the ∆ORF37/38 strain did not. Dexamethasone was used to confirm latent herpesvirus infection and reactivation. Dexamethasone injection increased viral DNA load in the brains of hamsters infected with the parental and ∆ORF38 strains, but not in those infected with the ∆ORF37/38 strain. Immunizing hamsters intranasally with the ∆ORF37/38 strain as a live vaccine produced a stronger immune response compared to the ∆ORF38 strain at the same dose. The hamsters demonstrated effective protection against a lethal challenge with the parental strain. This suggests that the deletion of ORF37/38 may effectively inhibit latent viral infection, reduce the neuropathogenicity of EHV-1, and induce a protective immune response.

From Human Perception of Good Practices to Horse (Equus Caballus) Welfare: Example of Equine-Assisted Activities.

Equine-assisted intervention (EAI) studies deal with clients, whereas very few studies focused on the effects on animals. EAI equids are also submitted to management, which influences their welfare. Management and working conditions depend on human decisions and perception. We gathered information through a survey about facilities managers' strategies (n = 51) and obtained direct information on management and working practices and their consequences on equids' welfare through an observational study (n = eight facilities, 174 equids). Differences in managers' perceptions of good management practices were related to the facility's involvement in EAI, e.g., increased awareness of equids' needs (housing and feeding), especially when EAI was the main activity. A detailed observational study on eight additional facilities confirmed that. Facility management profiles were paralleled by equids' welfare profiles. Clear correlates were found between management decisions and welfare consequences. One major factor influencing welfare and human-equid interactions appeared to be working modalities, with more EAI facilities practicing groundwork and bitless work. Facilities where equids were the most involved in mixed activities had the most equids with compromised welfare. Given EAI clients' particularities, conventional working modalities are less adapted and at risk of increased discomfort for equids. Overall, survey and observational approaches converged but some discrepancies (choice of equid type) appeared between the reported and observed prevalence.

Mule trains to mountain roads: the role of working mules in supporting resilient communities in the Himalayas.

Working equids play a central role in mountainous communities, but their work often goes unnoticed by the wider world, with sparse documentation of their role, value, or welfare - a state which often extends to their human counterparts. Communities living in the remote Manaslu Valley, Nepal, face a number of uncertainties, including extreme weather events due to the seasonal monsoon and, more recently, the construction of a new road network. Using semi-structured interviews, questionnaires, and Equine Assessment Research and Scoping (EARS) welfare assessments, we outline the specific role of pack mules in supporting the lives of local people, explain the nuanced links between human experience and mule welfare, and gain insight into how people living in this volatile environment manage uncertainty and risk. Mule work was felt to be the 'only option' for a sustainable livelihood for most mule owners although, in some cases, mules had enabled respondents to diversify their income. Mule owners with more husbandry experience did not own mules in more positive behavioural states, which may suggest a lack of generational knowledge and support networks. Short-term ongoing risks, such as the monsoon or unstable tracks, had a larger impact both financially and emotionally than the long-term but distant implications of the road construction. Mule owners must constantly balance the risk of working during the monsoon season, when conditions are treacherous but pay was higher, with losing valuable income but keeping themselves and their mules safe; they do, however, have a more mobile option for employment than non-owners. Mules enable a level of resilience and agility for communities living with constant uncertainty and change, which is only beginning to be recognised formally within the sustainable development sphere. Integration of animal welfare into the SDGs would allow humanitarian aid initiatives to strengthen support networks around working equids, which would greatly benefit the mules and humans alike.

Why Were Zebras Not Domesticated? A Review of Domesticability Traits and Tests of Their Role in Ungulate Domestications with Macroevolutionary Models.

Since Darwin, many evolutionary and behavioral researchers have considered the role of phenotypic traits that favor the domestication of nonhuman animals. Among such proposed traits are a species' social structure, level of intra- and interspecific agonistic interactions, sociosexual behaviors, parental strategies, reaction to humans, habitat preference, dietary habits, developmental trajectories, and utility to humans. However, little to no comparative phylogenetic evidence exists concerning the importance of these attributes for the domestication of animals. Moreover, rather than considering domestication as a dichotomous event (non-domesticated vs. domesticated), humans and their potential domesticates encountered numerous socioecological challenges/obstacles during the domestication process before reaching the stage of full domestication. The present study explored the influence of adult body mass, gregariousness, dietary breadth, and reaction to humans on the domestication process of ungulates. The phylogenetic comparative model revealed that capture myopathy (CM), as a proxy for reaction to humans, negatively and significantly influenced the domestication process. The present paper also explored the evolution of CM in equine species in response to the presence of large carnivoran predators during the Pleistocene. Ecologies that preserved most of the large carnivoran predators of equine species also featured more equine taxa with CM (e.g., zebras), which were thus less suitable for domestication.

Complete Blood Count and Biochemistry Reference Intervals for Healthy Adult Donkeys in the United States.

Previous hematologic and serum biochemistry reference interval (RI) values have been established for donkeys in various geographic regions, life-stages, or for specific donkey breeds. The last extensive investigation establishing RIs for adult donkeys in the United States (U.S.) was published over three decades ago. We aimed to establish updated robust RIs using a reference population of apparently healthy adult donkeys from across the U.S. Standard sized (n = 102), miniature (n = 17), and mammoth (n = 1) donkeys from four different states were enrolled, with 20% of the study population including donkeys captured directly from the wild in Death Valley National Park, CA. RIs were established in accordance with the American Society for Veterinary Clinical Pathology and Clinical Laboratory Standards Institute guidelines. The findings will assist practitioners with the interpretation of their complete blood count and biochemistry panel results in U.S. donkeys. This study also highlights a comparison of results for some important analytes in U.S. donkeys compared to U.S. horses and previously established donkey RIs.

What Is a Working Equid? Analysis of Current Terminology and a Suggested Definition.

Equids are engaged in myriad types of work across the world, with contexts and industries varying widely between high-, middle- and low-income countries as classified by the World Bank. Through a comprehensive abstract search and literature review, we examined the usage and context of terms associated with working equids in high-income countries (HICs), upper-middle-income countries (UMICs), lower-middle-income countries (LMICs), and low-income countries (LICs). Analysis showed that the search term used was significantly associated with World Bank country income classification. All search terms except two returned a significantly higher number of papers for higher-income countries compared to lower-income countries. The two exceptions were "working equid" and "draft equid", which returned significantly more papers for lower-income countries than higher-income countries. Both terms also returned papers from high-income and upper-middle-income countries, but these were always in the context of low-resource settings and rural or marginalised communities, of which we provide examples in the discussion. We propose that the term "working equid" should be used to describe "any equid engaged in physical labour that provides a significant or direct contribution to the economic livelihood, sustenance or support of the owner/user's family, typically within a low resource setting". Our review highlights the intricate interplay between socioeconomic factors and examines how equids are described in the literature. The lack of a universally accepted definition leads to challenges in policy formulation, resource allocation, and welfare standards development, particularly in LMICs and LICs where working equids are crucial to livelihoods. This study underscores the need for a more inclusive and comprehensive approach to terminology, advocating for future research to bridge linguistic and cultural gaps in understanding working equids globally. Such efforts are vital for equitable and effective welfare standards and policy development for these animals.

Natural grazing by horses and cattle promotes bird diversity in a restored European alluvial grassland.

Context: A challenge in grassland conservation is to maintain both the openness and the heterogeneity of the habitat to support the diversity of their animal communities, including birds-a taxon that is known to be sensitive to disturbance. An increasingly used management tool in European grassland conservation, especially in rewilding projects, is grazing by large herbivores such as horses and cattle. These grazers are believed to create and maintain patchy landscapes that promote diversity and richness of other species, but their influence on birds is often debated by conservationists, who raise concerns about the impact of disturbance by the grazers. Objectives: Our aim was to examine the relationship between the abundance and species richness of birds across four foraging guilds and the area utilization patterns of Highland cattle and Konik horses in an alluvial grassland in France. We also aimed to examine the influence of land cover and season on the spatial distribution, including abundance and species richness, of different bird guilds present in the grazed area. Methods: We used GPS-collars on all grazers and recorded their positions on an hourly basis over a study period of 1.5 years, assessing patterns of area usage. We counted birds weekly along three transects to describe their distribution within the grazed area and carried out land-cover surveys to describe the habitat. To assess how species richness and abundance of birds of different guilds were related to grazer density, season, and habitat characteristics, we used GAMM models in a spatially explicit framework. We also compared bird numbers at our main study site with a nearby non-grazed control area. Results: The number of birds in the grazed area was about twice the number in the non-grazed control area. Within the grazed area, the abundance of open-area foraging birds increased with increasing grazer density. The number of woodland-foraging birds was also positively correlated with grazer density but less so than open-area foraging birds. The number of individuals in the aerial and wetland bird guilds was not correlated with the density of grazers. Most bird species and individuals were observed on open landscapes scattered with woody patches and waterbodies, and on areas with moderate grazer density. Conclusions: Low-intensity grazing represents a potentially important management tool in creating heterogeneity in alluvial grasslands, thereby promoting suitable habitat for a diverse assemblage of bird species.

Development of a live attenuated vaccine candidate for equid alphaherpesvirus 1 control: a step towards efficient protection.

Equid alphaherpesvirus 1 (EqAHV1) is a viral pathogen known to cause respiratory disease, neurologic syndromes, and abortion storms in horses. Currently, there are no vaccines that provide complete protection against EqAHV1. Marker vaccines and the differentiation of infected and vaccinated animals (DIVA) strategy are effective for preventing and controlling outbreaks but have not been used for the prevention of EqAHV1 infection. Glycoprotein 2 (gp2), located on the envelope of viruses (EqAHV1), exhibits high antigenicity and functions as a molecular marker for DIVA. In this study, a series of EqAHV1 mutants with deletion of gp2 along with other virulence genes (TK, UL24/TK, gI/gE) were engineered. The mutant viruses were studied in vitro and then in an in vivo experiment using Golden Syrian hamsters to assess the extent of viral attenuation and the immune response elicited by the mutant viruses in comparison to the wild-type (WT) virus. Compared with the WT strain, the YM2019 Δgp2, ΔTK/gp2, and ΔUL24/TK/gp2 strains exhibited reduced growth in RK-13 cells, while the ΔgI/gE/gp2 strain exhibited significantly impaired proliferation. The YM2019 Δgp2 strain induced clinical signs and mortality in hamsters. In contrast, the YM2019 ΔTK/gp2 and ΔUL24/TK/gp2 variants displayed diminished pathogenicity, causing no observable clinical signs or fatalities. Immunization with nasal vaccines containing YM2019 ΔTK/gp2 and ΔUL24/TK/gp2 elicited a robust immune response in hamsters. In particular, compared with the vaccine containing the ΔTK/gp2 strain, the vaccine containing the ΔUL24/TK/gp2 strain demonstrated enhanced immune protection upon challenge with the WT virus. Furthermore, an ELISA for gp2 was established and refined to accurately differentiate between infected and vaccinated animals. These results confirm that the ΔUL24/TK/gp2 strain is a safe and effective live attenuated vaccine candidate for controlling EqAHV1 infection.

Assessment of in vitro digestibility and postdigestion peptide release of mare milk in relation to different management systems and lactation stages.

Mare milk has a unique protein composition that makes it a preferred option for adult and infant nutrition. Several functional properties have been attributed to this milk, but with little current evidence. In fact, knowledge on mare milk composition is still limited. In particular, studies addressing the performance of mare milk proteins during human gastrointestinal digestion are scarce, which limits the understanding of mare milk nutritional quality and functionality. For this reason, the present study describes the digestibility of mare milk proteins and the release of peptides as affected by management and lactation stage, factors which are known to affect milk composition. Mare milk samples from 3 different farms collected during 6 mo of lactation (n = 54) were subjected to a static in vitro gastrointestinal model to measure peptide release and protein digestibility. In the present study, a detailed description of protein and individual amino acid behavior during the digestion process was given. For the first time, digestion of the 2 equine ß-lactoglobulin isoforms (I and II) was described individually. In addition, it was found that lactation stage and management system can significantly affect protein digestibility and peptide release during gastrointestinal digestion of mare milk. Presumably, differences in the composition of mare milk influence the protein structure and enzyme accessibility, which might have an impact on digestion behavior. Although no specific bioactive peptides were identified, several precursors of previously described bioactive peptides were found. These findings could support the idea of mare milk as a food with added value.

Report on a Milking Mule: Milk Qualitative Characteristics during Lactation.

Despite their inability to reproduce naturally, mules can host embryos and be surrogate dams. The aim of this investigation was to increase our knowledge of the qualitative composition of mule's milk and its variations throughout the whole lactation period-namely, from 6 h after foaling to 180 days in milk (DIM). Milk was obtained from a mule dam that had foaled after receiving a mule embryo transfer. For each sample, the gross, mineral, and fatty acid composition was evaluated. The average quality of the mule milk was as follows: protein 1.97 g 100 mL-1, fat 0.90 g 100 mL-1, and ash 0.39 g 100 mL-1. Saturated fatty acids made up, on average, 50.00 g 100 g-1 of fat. Monounsaturated and polyunsaturated fatty acids made up half of the total fatty acid content (31.80 g 100 g-1 and 18.2 g 100 g-1 of fat, respectively). Linoleic acid and linolenic acid were the main polyunsaturated fatty acids in the milk. The milk composition changed throughout lactation. Dry matter, protein, fat, and ash decreased significantly from early lactation (6 h to 14 DIM). The n3 polyunsaturated fatty acids decreased at the end of lactation. The changes in milk composition throughout lactation are probably due to adaptations to the growth requirements of the foal.

Cardiac Disease Related to Primary Hyperthyroidism in a 20-Year-Old Mule.

Primary hyperthyroidism is a rarely diagnosed endocrinopathy in equids and there have been no previous reports of structural and functional cardiac changes associated with hyperthyroidism in these species. This case report investigates a 20-year-old mule gelding that presented for a three-month history of thin body condition despite polyphagia, with a heart murmur and elevated free and total thyroid hormone concentrations. On presentation, physical exam revealed a body condition score of two out of nine, persistent tachycardia, pansystolic heart murmur and firm bilateral ventral proximal cervical masses. Bloodwork confirmed markedly elevated free T4, total T4 and T3 concentrations. Echocardiogram demonstrated left ventricular concentric hypertrophy with increased ventricular and atrial systolic function. Bilateral thyroidectomy was performed under standing sedation without complications. Histopathology demonstrated adenocarcinoma of the left thyroid gland and multiple adenomas with osseous metaplasia within the right thyroid. The mule was supplemented with levothyroxine sodium two weeks post-op after a thyroid panel demonstrated undetectable concentrations. Polyphagia resolved following surgery and the mule began gaining weight. Echocardiographic changes improved but did not resolve at two years post-operative. Continued bi-annual follow up and monitoring of thyroid levels was recommended. This case represents the first documentation of hemodynamically relevant cardiac remodeling in an equid associated with primary hyperthyroidism.

Unmasking the ongoing challenge of equid herpesvirus- 1 (EHV-1): A comprehensive review.

Equid herpesviruses (EHVs) are a group of highly impactful viral pathogens that affect horses, presenting a substantial risk to the global equine industry. Among these, equid herpesvirus-1 (EHV-1) primarily causes respiratory infections. However, its ability to spread to distant organs can lead to severe consequences such as abortion and neurological diseases. These viruses can enter a dormant phase, with minimal activity, and later reactivate to trigger active infections at any time. Recently, there has been a notable rise in the prevalence of a particularly devastating strains of EHV-1 known as equid herpesviral myeloencephalopathy (EHM). In the light of dynamic nature of EHV-1, this review provides a thorough overview of EHV-1 and explores how advances in viral biology affect the pathophysiology of viral infection. The information presented here is crucial for understanding the dynamics of EHV-1 infections and creating practical plans to stop the virus's global spread among equid populations.

Transvaginal aspiration, oocyte maturation and production of blastocysts and a pregnancy via ICSI in an endangered breed: Baudet du Poitou donkey.

Two Poitou donkey jennies were presented for clinical oocyte recovery and embryo production via intracytoplasmic sperm injection (ICSI). Both jennies underwent transvaginal ultrasound-guided follicle aspiration on two occasions. Recovered oocytes were held overnight then placed into maturation culture, using standard methods for mare oocytes. On the first replicate for both jennies, the oocytes were divided into two groups; one group was denuded and examined at 30 h culture (standard culture duration for mare oocytes) and the second was denuded and examined at 36 h culture. No oocytes with polar bodies were observed at either time. The oocytes were maintained in maturation culture until 46 h, at which time oocytes with polar bodies were observed. Semen was then prepared; oocytes underwent ICSI approximately 48 h after being placed into maturation culture. On the second replicate for both jennies, oocytes were cultured for maturation for 42 h, then denuded and subjected to ICSI at 46 h. Sperm preparation, injection and embryo culture were performed as for mare oocytes. Blastocyst rates per injected oocyte were 8/19 (42 %) overall, being 4/12 and 4/7 for the first and second TVAs, respectively. Blastocysts were vitrified. Three blastocysts were warmed and transferred to Poitou donkey jenny recipients. One embryonic vesicle was visualized on ultrasonography on embryo Day 12, which increased in size on Day 13 but was not present when examined on Day 14. These results demonstrate that oocyte recovery and ICSI are efficient for production of Poitou donkey blastocysts. To the best of our knowledge, this is the first report of production of blastocysts via ICSI in the Poitou donkey, and the first report of transfer of ICSI-produced embryos in the donkey. Further work is needed on factors affecting pregnancy after embryo transfer in the donkey.

A Screening Study Identified Decitabine as an Inhibitor of Equid Herpesvirus 4 That Enhances the Innate Antiviral Response.

Equid herpesvirus 4 (EHV-4) is a common respiratory pathogen in horses. It sporadically induces abortion or neonatal death. Although its contribution in neurological disorders is not clearly demonstrated, there is a strong suspicion of its involvement. Despite preventive treatments using vaccines against EHV-1/EHV-4, the resurgence of alpha-EHV infection still constitutes an important threat to the horse industry. Yet very few studies have been conducted on the search for antiviral molecules against EHV-4. A screening of 42 antiviral compounds was performed in vitro on equine fibroblast cells infected with the EHV-4 405/76 reference strain (VR2230). The formation of cytopathic effects was monitored by real-time cell analysis (RTCA), and the viral load was quantified by quantitative PCR. Aciclovir, the most widely used antiviral against alpha-herpesviruses in vivo, does not appear to be effective against EHV-4 in vitro. Potential antiviral activities were confirmed for eight molecules (idoxuridine, vidarabine, pritelivir, cidofovir, valganciclovir, ganciclovir, aphidicolin, and decitabine). Decitabine demonstrates the highest efficacy against EHV-4 in vitro. Transcriptomic analysis revealed the up-regulation of various genes implicated in interferon (IFN) response, suggesting that decitabine triggers the immune antiviral pathway.

First identification and isolation of equine herpesvirus type 1 in aborted fetal lung tissues of donkeys.

BACKGROUND: Equine herpesvirus type 1 (EHV-1) is commonly associated with horse abortion. Currently, there are no reported cases of abortion resulting from EHV-1 infection in donkeys. RESULTS: This was the first survey-based study of Chinese donkeys. The presence of EHV-1 was identified by PCR. This survey was conducted in Chabuchar County, North Xinjiang, China, in 2020. A donkey EHV-1 strain (Chabuchar/2020) was successfully isolated in MDBK cells. Seventy-two of 100 donkey sera were able to neutralize the isolated EHV-1. Moreover, the ORF33 sequence of the donkey-origin EHV-1 Chabuchar/2020 strain showed high levels of similarity in both its nucleotide (99.7‒100%) and amino acid (99.5‒100%) sequences, with those of horse EHV-1 strains. EHV-1 Chabuchar/2020 showed significant consistency and was classified within cluster 1 of horse EHV-1 strains. Further, analysis of the expected ORF30 nucleotide sequence revealed that donkey EHV-1 strains contained guanine at position 2254, resulting in a change to aspartic acid at position 752 of the viral DNA polymerase. Therefore, these strains were classified as horse neuropathogenic strains. Lastly, a phylogenetic tree was constructed using the partial ORF68 nucleotide sequences, showing that the identified donkey EHV-1 strain and the EHV-1 strain found in aborted Yili horses in China comprised a novel independent VIII group. CONCLUSION: This study showed the first isolation and identification of EHV-1 as an etiological agent of abortions in donkeys. Further analysis of the ORF33, ORF30, and ORF68 sequences indicated that the donkey EHV-1 contained the neuropathogenic genotype of strains in the VIII group. It is thus important to be aware of EHV-1 infection in the donkey population, even though the virus has only been identified in donkey abortions in China.

Modulation of Equid Herpesvirus-1 Replication Dynamics In Vitro Using CRISPR/Cas9-Assisted Genome Editing.

(1) Background: equid alphaherpesvirus-1 (EHV-1) is a highly contagious viral pathogen prevalent in most horse populations worldwide. Genome-editing technologies such as CRISPR/Cas9 have become powerful tools for precise RNA-guided genome modifications; (2) Methods: we designed single guide RNAs (sgRNA) to target three essential (ORF30, ORF31, and ORF7) and one non-essential (ORF74) EHV-1 genes and determine their effect on viral replication dynamics in vitro; (3) Results: we demonstrated that sgRNAs targeting essential lytic genes reduced EHV-1 replication, whereas those targeting ORF74 had a negligible effect. The sgRNAs targeting ORF30 showed the strongest effect on the suppression of EHV-1 replication, with a reduction in viral genomic copy numbers and infectious progeny virus output. Next-generation sequencing identified variants with deletions in the specific cleavage site of selective sgRNAs. Moreover, we evaluated the combination between different sgRNAs and found that the dual combination of sgRNAs targeting ORF30 and ORF7 significantly suppressed viral replication to lower levels compared to the use of a single sgRNA, suggesting a synergic effect; (4) Conclusion: data demonstrate that sgRNA-guided CRISPR/Cas9 can be used to inhibit EHV-1 replication in vitro, indicating that this programmable technique can be used to develop a novel, safe, and efficacious therapeutic and prophylactic approach against EHV-1.

"We forgot about the donkeys!" An institutional analysis of the shift in animal welfare from direct implementation towards advocacy-based programming.

Shifts from direct implementation to advocacy-based programming have been documented across many non-governmental organisation (NGO) sectors, including animal welfare. Semi-structured interviews with 32 staff from different positions within animal welfare NGOs explored recent programming changes. Maintaining a balance between direct implementation and advocacy-based activities emerged as a strong theme. The findings suggest that risks are associated with both the direct implementation status quo and transitioning to an advocacy-based focus. Risks of the former include treating symptoms rather than root causes of welfare problems. Organisational change can be disruptive and necessitates realignment of core competences, in turn influencing NGO mission. Identified risks of transition include loss of individuals whose values fail to align with new programming directions, increased upwards accountability requirements for accessing institutional donors and difficulties when phasing out direct implementation approaches. Whilst having to be dynamic, NGOs need to evaluate the risks associated with programming decisions, considering their vision, mission and staff identity in order to ensure that welfare programming is as effective as possible.

A field approach to observing changes in behavioural welfare indicators over 2 years in stabled horses.

In stabled horses, several behaviours are considered to be important indicators of the state of welfare at the individual level: stereotypies, aggressive behaviours towards humans and the "withdrawn", alert, sternal, lateral and observation postures. Until now, these behaviours have been extensively studied in relation to different horse management practices. However, little is known about their changes or consistency over time. This study aimed to investigate differences in the expression of these behaviours assessed on 44 stabled horses during an initial 3-month period and then again on the same individuals 2 years later. Out of the six behaviours studied, two showed significant differences between the 2 years with medium effect sizes: the levels of aggressiveness towards humans increased (Wilcoxon signed-rank test: V = 65, P = 0.005) and those of recumbent rest during the day decreased (V = 416, P < 0.001) over time. The results also suggested limited evidence of major changes over time in the expression of stereotypies, the "withdrawn", alert and observation postures. However, the principal component analyses carried out on all the behaviours showed that alert and observation postures may slightly decrease over time for some individuals. Regarding stereotypies and the "withdrawn" posture, the results mainly suggested a change at the individual level in the expression of these behaviours over time. This study provided new insights into the dynamic nature of several behaviours when the horses' living conditions are not fundamentally altered and raised hypotheses about the state of welfare of stabled individuals over a 2-year period.