United Kingdom

The control of equid gastrointestinal nematodes in developed countries, in particular the cyathostomins, is threatened by high levels of anthelmintic resistance. In recent years, there has been increasing interest in the evaluation of traditional ‘ethnoveterinary’ medicines as alternatives to chemical anthelmintics. The cysteine proteinases (CPs), a group of enzymes derived from fruits such as papaya (Carica papaya), pineapple (Ananas comosus) and figs (Ficus spp.), have shown good efficacy against adult stages of a range of parasitic nematodes, in vitro and in vivo. The efficacy of CPs against cyathostomins remains to be explored. In this study, the efficacy of a crude preparation of CPs, papaya latex supernatant (PLS), against the free-living stages of cyathostomins was evaluated using two in vitro tests, the egg hatch test (EHT) and the larval migration inhibition test (LMIT). It was demonstrated that PLS had a potent effect in the EHT, with EC-50 values in the range of 0.12-0.22 μM. At concentrations above 6.25 μM the eggs did not develop, below this concentration the L1 developed but they lost integrity of the cuticle upon hatching. These effects were inhibited by pre-incubation of PLS with the CP inhibitor L-trans-epoxysuccinyl-L-leucylamido-(4-guanidino butane) (E64), indicating that CPs were responsible for the anti-parasitic activity. A dose-dependent inhibition of migration of third stage larvae (L3) in the LMIT was demonstrated at higher concentrations of PLS, with EC-50 values in the range of 67.35-106.31 μM. Incubation of PLS with E64 prior to use in the LMIT did not reverse the anti-migratory effect, suggesting that CPs were not responsible for the reduced migration of cyathostomin L3 and that PLS also contains an additional active compound. This is the first report of PLS and/or CPs showing activity against the free-living stages of a parasitic helminth. In addition, it suggests that cyathostomins are highly sensitive to the effects of CPs and further evaluation of their efficacy against parasitic stages and in vivo are strongly indicated.

Anthelmintic resistance is a global problem that threatens sustainable control of the equine gastrointestinal cyathostomins (Phylum Nematoda; Superfamily Strongyloidea). Of the three novel anthelmintic classes that have reached the veterinary market in the last decade, none are currently licenced in horses, hence current control regimens focus on prolonging the useful lifespan of licenced anthelmintics. This approach would be facilitated by knowledge of the resistance mechanisms to the most widely used anthelmintics, the macrocyclic lactones (ML). There are no data regarding resistance mechanisms to MLs in cyathostomins, although in other parasitic nematodes, the ABC transporters, P-glycoproteins (P-gps), have been implicated in playing an important role. Here, we tested the hypothesis that P-gps are, at least in part, responsible for reduced sensitivity to the ML ivermectin (IVM) in cyathostomins; first, by measuring transcript levels of pgp-9 in IVM resistant versus IVM sensitive third stage larvae (L3) pre-and post-IVM exposure in vitro. We then tested the effect of a range of P-gp inhibitors on the effect of IVM against the same populations of L3 using the in vitro larval development test (LDT) and larval migration inhibition test (LMIT). We demonstrated that, not only was pgp-9 transcription significantly increased in IVM resistant compared to IVM sensitive L3 after anthelmintic exposure (p < 0.001), but inhibition of P-gp activity significantly increased sensitivity of the larvae to IVM in vitro, an effect only observed in the IVM resistant larvae in the LMIT. These data strongly implicate a role for P-gps in IVM resistance in cyathostomins. Importantly, this raises the possibility that P-gp inhibitor-IVM combination treatments might be used in vivo to increase the effectiveness of IVM against cyathostomins in Equidae.

Hyperlipaemia is a disease resulting from excessive mobilisation of triglyceride stores such that plasma clearance processes become overwhelmed. Consequently increased plasma triglyceride concentrations (>5.6 mmol/l), visibly cloudy plasma and a sick, anorexic or hypophagic subject follow. Epidemiological studies have identified many predisposing and triggering factors and the attentive carer or veterinary surgeon should be alert to such risk factors so that the disease can be prevented, or at least identified and treated at an early stage. Hyperlipaemic subjects are invariably in a negative energy balance, and nutritional management therefore plays a central role in both the prevention and resolution of the disease.

The domestic donkey is descended from wild asses and has evolved to live in some of the most inhospitable places on earth. Little research has been carried out to address the specific needs of the donkey, which has traditionally been viewed as a small horse. The donkey is different from the horse in many ways; of particular note is its ability to thrive on highly fibrous feeds. This article discusses the nutritional requirements of donkeys and how dental disease may play a role in determining their nutritional requirements.

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.

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

Laminitis is a painful disease of equines. Radiographic and post-mortem evaluations of feet are often an important part of welfare investigations, and professional opinions by veterinarians are necessary in resulting legal cases. Any difference in measurements between the two modalities can cause uncertainty, potentially affecting the legal decision.

Objectives

To quantify the difference between radiographic and postmortem pre-mortem vs. post-mortem effects.

Study design

Case series.

Methods

Seven donkeys with laminitis confirmed via standard workup, euthanased for reasons unrelated to the study, were selected. Weight-bearing and non-weight-bearing lateral radiographs were taken of both front feet within 24 h pre-mortem. Feet were removed and sagitally sectioned between 48 and 72 h post-mortem. Lateral radiographs were taken of the feet immediately following sectioning. Founder distance and rotation were evaluated at each time point and compared using paired t-tests (P < 0.05).

Results

Compared with pre-mortem weight-bearing radiographs, nonweight-bearing feet had a decreased founder distance and decreased rotation. Compared with pre-mortem non-weight- bearing radiographs, post-mortem feet had increased rotation and no change in founder distance. There were no significant differences between post-mortem direct measurements and post- mortem radiographs. Compared with standard weight-bearing radiographs, post-mortem measurements had a decreased founder distance and increased rotation.

Main limitations

Small sample size. Further samples are needed to confirm these initial conclusions.

Conclusions

Measurements of post-mortem feet have a decreased founder distance and an increased rotation compared with standard radiographic images. Changes in founder distance are seen due to changes in weight-bearing. Changes in rotation are seen post-mortem, and can be explained by autolysis of the laminae and/or rigor mortis causing tendon contracture. Most studies have focused on indications and severity of laminitis in living animals using radiographs: postmortem measurements should therefore be interpreted with caution.

Competing interests

None declared.

Ethical animal research

Approved by The Donkey Sanctuary. Donkeys were owned by The Donkey Sanctuary and were used with consent.

Sources of funding

The Donkey Sanctuary.

This article provides a review of hoof anatomy and care in donkeys and mules. Hoof disease is a major cause of poor welfare and mortality globally. Problems associated with hoof disease are discussed in the context of behavior, diet, treatment, and prevention. The most common conditions encountered are discussed, including laminitis, the overgrown unbalanced hoof, white line disease, flexural deformities, and other significant issues. Differences between donkey and horse hoof anatomy are described.

Published online ahead of print.

Background

There are clear differences between donkeys and horses in their evolutionary history, physiology, behaviour and husbandry needs. Donkeys are often kept in climates that they are not adapted to and as such may suffer impaired welfare unless protection from the elements is provided.

Objectives

To compare some of the hair coat properties of donkeys, mules and horses living outside, throughout the year, in the temperate climate of the UK.

Study design

Longitudinal study.

Methods

Hair samples were taken from 42 animals: 18 donkeys (4 females, 14 males), 16 horses (6 females, 10 males) and eight mules (5 females, 3 males), in March, June, September and December. The weight, length and width of hair were measured, across the four seasons, as indicators of the hair coat insulation properties.

Results

Donkeys’ hair coats do not significantly differ across the seasons. All three measurements of the insulation properties of the hair samples indicate that donkeys do not grow a winter coat and that their hair coat was significantly lighter, shorter and thinner than that of horses and mules in winter. In contrast, the hair coats of horses changed significantly between seasons, growing thicker in winter.

Main limitations

The measurements cover only a limited range of features that contribute to the thermoregulation of an animal. Further research is needed to assess shelter preferences by behavioural measures, and absolute heat loss via thermoimaging.

Conclusions

Donkeys, and to a lesser extent mules, appear not to be as adapted to colder, wet climates as horses, and may therefore require additional protection from the elements, such as access to a wind and waterproof shelter, in order for their welfare needs to be met.

A review of common gastrointestinal disorders of donkeys and mules is presented. Clinically relevant aspects of donkey behavior, anatomy, and physiology are highlighted. Diagnosis, management, and treatment of conditions affecting the gastrointestinal tract from stomach to rectum, including liver and pancreas, are discussed.

Published online ahead of print.