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Multidrug resistance (MDR) is already occurring among some equids in India. Donkeys and mules are a mobile species moving between regions and international borders, often populating areas of India where private community pharmacies, or medical stores, are the primary healthcare provider for both humans and animals. This article highlights how the capacities of drug retail outlet workers might affect their antibiotic dispensing practices, particularly in relation to donkeys and mules, in order to consider how this might impact the development of antimicrobial resistance (AMR) on a wider scale. A mixed-methods approach was implemented using patient simulation method (n = 28), semi-structured interviews (SSIs) (n = 23), focus group discussions (FGDs) with veterinary practitioners and non-governmental organisation animal health workers (n = 2 FGDs), and participant observation. Fewer than 48 per cent of drug retail outlet workers admitted to having had any formal training in pharmaceuticals at all, while 78 per cent reported having no formal training in animal-related pharmaceuticals. Moreover, 35 per cent of all participants sold antibiotics without a prescription, unprompted and without specifically being asked for antibiotics. Of the antibiotics dispensed, only 21 per cent were correctly dispensed for the symptoms presented, and all dosages dispensed were incorrect (underdosed). Furthermore, 43 per cent of drug retail outlet workers interviewed believe that some antibiotics can be legally dispensed without a prescription. Equine owners in northern India are frequently being sold antibiotics without a prescription and, in most cases, with incorrect diagnoses, treatment choice, and dosage. A substantial gap in capacities exists amongst Drug Retail Outlet (DRO) workers, with few being sufficiently qualified or trained to dispense antibiotics to animal owners. The study highlights the need for further training of private DRO workers as well as knowledge extension and awareness training for both DRO workers and animal owners regarding antimicrobial resistance and its potential impact upon livelihoods. It also illustrates the need to identify a balance whereby greater enforcement of regulation at all levels is implemented, while at the same time maintaining sufficient access to medicine for rural populations.

Background

Equine gut microbiology studies to date have primarily focused on horses and ponies, which represent only one of the eight extant equine species. This is despite asses and mules comprising almost half of the world’s domesticated equines, and donkeys being superior to horses/ponies in their ability to degrade dietary fiber. Limited attention has also been given to commensal anaerobic fungi and archaea even though anaerobic fungi are potent fiber degrading organisms, the activity of which is enhanced by methanogenic archaea. Therefore, the objective of this study was to broaden the current knowledge of bacterial, anaerobic fungal and archaeal diversity of the equine fecal microbiota to multiple species of equines. Core taxa shared by all the equine fecal samples (n = 70) were determined and an overview given of the microbiota across different equine types (horse, donkey, horse × donkey and zebra).

Results

Equine type was associated with differences in both fecal microbial concentrations and community composition. Donkey was generally most distinct from the other equine types, with horse and zebra not differing. Despite this, a common bacterial core of eight OTUs (out of 2070) and 16 genus level groupings (out of 231) was found in all the fecal samples. This bacterial core represented a much larger proportion of the equine fecal microbiota than previously reported, primarily due to the detection of predominant core taxa belonging to the phyla Kiritimatiellaeota (formerly Verrucomicrobia subdivision 5) and Spirochaetes. The majority of the core bacterial taxa lack cultured representation. Archaea and anaerobic fungi were present in all animals, however, no core taxon was detected for either despite several taxa being prevalent and predominant.

Conclusions

Whilst differences were observed between equine types, a core fecal microbiota existed across all the equines. This core was composed primarily of a few predominant bacterial taxa, the majority of which are novel and lack cultured representation. The lack of microbial cultures representing the predominant taxa needs to be addressed, as their availability is essential to gain fundamental knowledge of the microbial functions that underpin the equine hindgut ecosystem.

Working equids rely on sound, balanced hooves, but data describing the typical morphology of the legs and feet of working donkeys are currently lacking. To address this gap in knowledge, the front and hind feet of twenty healthy working donkeys were measured and compared. Hoof width, weight-bearing lengths, heel width, dorsal hoof wall length and lateral and medial heel length of the hoof wall were determined, as well as toe angle, heel angle, hoof pastern axis, coronary band angle and a measure of ‘ground surface size’. Viewed from the ground surface of the foot, front feet were more rounded and significantly larger than hind feet. Measures of medial-lateral balance and toe-heel angle ratio were within the recommended healthy guidelines for horses. Hoof pastern axis was broken forward for the studied animals, which supports previous research suggesting that a broken forward hoof pastern axis is normal for donkeys, although further study would be required to confirm whether this conformation is natural. Significant correlations were found between estimated body mass and hoof width in both the front and hind feet. These measurements provide valuable insight into the relationship between hoof and body characteristics, which may aid the development of guidelines for the trimming and management of working donkey hooves. Further study is, however, advised to confirm natural hoof conformation.

Horses and donkeys belong to the genus Equus, but important differences exist between the species, many of which affect their management and welfare. This study compared skull morphology between horses and donkeys. Horse (n = 14) and donkey (n = 16) heads were obtained post-mortem, sectioned sagittally close to the midline, and photographed for subsequent measurement of various skull structures. Skull, cranial, nasal, and profile indices were calculated for topographical comparisons between the species. The olfactory bulb area (OBA), OB pitch (the angle between the hard palate and the OB axis), and whorl location (WL) were also measured. A General Linear Model determined the main effect of species with Sidak’s multiple comparisons of species’ differences among the various measurements. There was no species difference in cranial or nasal indices (p > 0.13), but donkeys had a larger cranial profile than horses (p < 0.04). Donkeys had a smaller OBA (p < 0.05) and a steeper OB pitch (p < 0.02) than horses. The WL corresponded to the level of the OB in horses but was extremely rostral in donkeys (p < 0.0001). These results show clear differentiation in skull morphology between horses and donkeys. This may be useful in validating other physiological and behavioural differences between horses and donkeys.

Background

The involvement of Besnoitia bennetti in skin pathologies was investigated in a series of 20 donkeys from the Donkey Sanctuary in England, in the 2013–2019 period.

Methods

The initial histopathological finding of Besnoitia cysts in skin lumps that were presumed to be sarcoids in 2013 triggered our cognisance of this parasite and resulted in identification of a total of 20 cases. Histopathological examination of surgical biopsy samples collected from 8 live donkeys and tissue specimens from 12 deceased donkeys at post-mortem examination revealed the presence of Besnoitia cysts in all 20 donkeys. The indirect fluorescent antibody test (IFAT) and immunoblotting analysis showed the presence of anti-Besnoitia antibodies in archived serum samples from 4 deceased donkeys. Additionally, infection was evidenced in one live donkey based on IFAT and immunoblot analysis of tissue fluid of a dermal mass containing Besnoitia cysts, and real-time (RT)-PCR analysis and microsatellite genotyping of DNA isolated from the tissue of the same dermal mass confirmed the infection specifically as B. bennetti.

Results

Both serological and microsatellite analyses confirmed the aetiology to be B. bennetti. Our findings suggested that in cases of skin masses such as sarcoids, the suspicion of B. bennetti infection should be borne in mind even when clinical and histopathology examination results are negative in order to avoid misdiagnosis.

Conclusions

This case series documents, to our knowledge, the first report of B. bennetti infection in donkeys in the UK, indicating that donkey besnoitiosis has become noteworthy in the UK. Further investigations of the occurrence, epidemiological characteristics, and clinical manifestations of B. bennetti infection in donkeys and other equids are warranted.

The assessment of animal welfare poses numerous challenges, yet an emerging approach is the consolidation of existing knowledge into new frameworks which can offer standardised approaches to welfare assessment across a variety of contexts. Multiple tools exist for measuring the welfare of equids, but such tools have typically been developed for specific contexts. There is no ‘one size fits all’ which means that resulting datasets are generally non-comparable, creating a barrier to knowledge-sharing and collaboration between the many organisations working to improve equid welfare around the globe. To address this, we developed the Equid Assessment, Research and Scoping (EARS) tool, which incorporates pre-existing validated welfare assessment methods alongside new welfare indicators to deliver a larger and more comprehensive series of welfare indicators than currently exists, creating a single resource that can be used to assess equid welfare in any context. We field-trialled three welfare assessment protocols within the EARS tool, and applied these to welfare assessment of equids in a variety of contexts across nineteen countries. The EARS tool proved a useful, versatile and rapid method for collecting welfare assessment data and we collected 7464 welfare assessments in a period of fifteen months. We evaluate the EARS tool and provide ideas for future development.

Recently, the need for a more holistic approach to welfare assessment has been highlighted. This is particularly pertinent in the case of working equids who provide vital support for human livelihoods, often in low- to middle-income countries, yet suffer from globally low standards of welfare. This study aimed to provide insight into the welfare status and traditional use of working equids in rural Western European communities using the new EARS welfare tool, designed to provide a broad view of the welfare of working equids and the context in which they are found. Other questions on the topics of equid management practices, social transmission of expertise, environmental stressors, and traditions, alongside physical and behavioural welfare assessments were also included to explore the impact of these wide-ranging factors on an understudied population of working equids. The protocol was trialled on 60 working equid owners from communities in Portugal and Spain where, despite the decline in traditional agricultural practices and livestock keeping, donkeys and mules remain working animals. Many owners stated that the help donkeys provided was invaluable, and donkeys were considered to be important for both farming and daily life. However, participants also recognised that the traditional agricultural way of life was dying out, providing insights into the traditional practices, community structure, and beliefs of equid owners. Questions investigating the social networks and social transfer of information within the villages were effective in finding local sources of equid knowledge. Overall, welfare was deemed fair, and the protocol enabled the identification of the most prevalent welfare problems within the communities studied, in this case obesity and the use of harmful practices. The findings suggest that the new protocol was feasible and detail how contextual factors may influence equid welfare. Increasing understanding of the cultural context, social structure, and attitudes within a community, alongside more traditional investigations of working practices and animal management, may, in the future, help to make equid welfare initiatives more effective.

Non-governmental organizations (NGOs) work across the globe to improve the welfare of working equids. Despite decades of veterinary and other interventions, welfare issues persist with equids working in brick kilns. Engagement with all stakeholders is integral to creating abiding improvements to working equid welfare as interventions based purely on reactive measures fail to provide sustainable solutions. Equid owners, particularly those in low to middle-income countries (LMICs), may have issues such as opportunity, capacity, gender or socio-economic status, overriding their ability to care well for their own equids. These “blind spots” are frequently overlooked when organizations develop intervention programs to improve welfare. This study aims to highlight the lives of the poorest members of Indian society, and will focus on working donkeys specifically as they were the only species of working equids present in the kilns visited. We discuss culture, status, religion, and social influences, including insights into the complexities of cultural “blind spots” which complicate efforts by NGOs to improve working donkey welfare when the influence of different cultural and societal pressures are not recognized or acknowledged. Employing a mixed-methods approach, we used the Equid Assessment Research and Scoping (EARS) tool, a questionnaire based equid welfare assessment tool, to assess the welfare of working donkeys in brick kilns in Northern India. In addition, using livelihoods surveys and semi-structured interviews, we established owner demographics, socioeconomic status, ethnicity, religion and their personal accounts of their working lives and relationships to their donkeys. During transcript analysis six themes emerged: caste, ethnicity, inherited knowledge; social status, and impacts of ethnic group and caste; social status and gender; migration and shared suffering; shared suffering, compassion; religious belief, species hierarchy. The lives led by these, marginalized communities of low status are driven by poverty, exposing them to exploitation, lack of community cohesion, and community conflicts through migratory, transient employment. This vulnerability influences the care and welfare of their working donkeys, laying bare the inextricable link between human and animal welfare. Cultural and social perspectives, though sometimes overlooked, are crucial to programs to improve welfare, where community engagement and participation are integral to their success.

Equids fulfil many different roles within communities. In low- to middle-income countries (LMICs), in addition to providing a source of income, equids also provide essential transport of food, water, and goods to resource-limited and/or isolated communities that might otherwise lack access. The aim of this investigation was to understand the welfare conditions that donkeys, mules, and horses are exposed to whilst working in Nepalese brick kilns. To understand the welfare conditions of equids in Nepalese brick kilns, the Welfare Aggregation and Guidance (WAG) tool in conjunction with the Equid Assessment, Research and Scoping (EARS) tool was used to understand the health, behaviour, nutrition, living and working conditions in brick kilns. Further analysis of individual EARS responses focused on key indicator questions relating to demographic information was used to investigate specific areas of welfare concern and attitudes of handlers towards their equids. Trained staff carried out welfare assessments between December 2018 and April 2019. The information gathered using the EARS tool was summarised using the WAG tool to pinpoint areas of welfare concern and suggest possible strategies to mitigate poor welfare conditions and suggest areas to improve the welfare of equids. Overall, the results indicate that to improve the welfare of equids working in Nepalese brick kilns, there should be better provision of clean water, both when working and stabled, equipment should be removed and shade provided during rest periods, with improvements made to housing to allow the equids to rest and recuperate when not working. Further work should also focus on collaborating with owners and equid handlers to improve their attitudes and practices towards their equids. Such improvements can be implemented via training of equid handlers and kiln owners whilst using the EARS and WAG tools to provide a sound basis on which to monitor the effectiveness and impact of education programs on equid welfare.