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 phylaKiritimatiellaeota(formerlyVerrucomicrobiasubdivision 5) andSpirochaetes. 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.

Background: Compared to horses and ponies, donkeys have increased degradation of dietary fiber. The longer total mean retention time of feed in the donkey gut has been proposed to be the basis of this, because of the increased time available for feed to be acted upon by enzymes and the gut microbiota. However, differences in terms of microbial concentrations and/or community composition in the hindgut may also underpin the increased degradation of fiber in donkeys. Therefore, a study was conducted to assess if differences existed between the fecal microbiota of pony, donkey and hybrids derived from them (i.e. pony × donkey) when fed the same forage diet.Results: Fecal community composition of prokaryotes and anaerobic fungi significantly differed between equine types. The relative abundance of two bacterial genera was significantly higher in donkey compared to both pony and pony x donkey:Lachnoclostridium10 and ‘probable genus 10’ from theLachnospiraceaefamily. The relative abundance ofPiromyceswas significantly lower in donkey compared to pony × donkey, with pony not significantly differing from either of the other equine types. In contrast, the uncultivated genus SK3 was only found in donkey (4 of the 8 animals). The number of anaerobic fungal OTUs was also significantly higher in donkey than in the other two equine types, with no significant differences found between pony and pony × donkey. Equine types did not significantly differ with respect to prokaryotic alpha diversity, fecal dry matter content or fecal concentrations of bacteria, archaea and anaerobic fungi.Conclusions: Donkey fecal microbiota differed from that of both pony and pony × donkey. These differences related to a higher relative abundance and diversity of taxa with known, or speculated, roles in plant material degradation. These findings are consistent with the previously reported increased fiber degradation in donkeys compared to ponies, and suggest that the hindgut microbiota plays a role. This offers novel opportunities for pony and pony × donkey to extract more energy from dietary fiber via microbial mediated strategies. This could potentially decrease the need for energy dense feeds which are a risk factor for gut-mediated disease.

Free faecal water (FFW) in equines results in pollution of the hindquarters and tail and can also involve clinical signs. Though the cause of FFW is unknown, it was hypothesized that it may involve the gut microbiota. This hypothesis was addressed as follows. First, the faecal prokaryotic community composition of horses suffering from FFW relative to healthy controls (n = 10) was compared. Second, FFW horses were treated with a standardised faecal microbiota transplantation (FMT) protocol (n = 10), followed by assessment of FFW symptom severity and faecal prokaryotic community composition over a follow-up period of 168 days. No significant differences were found in the faecal microbiota composition of FFW horses compared to healthy controls (p > 0.05). Relative to before FMT, FFW symptom severity decreased in affected horses 14 days after FMT (p = 0.02) and remained decreased for the remainder of the study (p < 0.02). However, individual animal responses to FMT varied. FMT had no effect on FFW horse faecal prokaryotic community composition in terms of alpha or beta diversity. Alpha diversity of the donor inocula used in the FMT was always lower than that of the faecal microbiota of the FFW treated horses (p < 0.001). In conclusion, whilst findings indicate FFW horses do not have an altered hindgut microbiota, some horses that received FMT had a temporary alleviation of FFW symptom severity without causing changes in the faecal microbiota. Future studies using controls are now needed to confirm the effectiveness of FMT to treat FFW.

Equine squamous gastric disease (ESGD) is common in horses and poses a serious welfare problem. Several risk factors have been identified and ESGD is routinely treated with omeprazole. Fourteen mares, previously used as embryo recipients and diagnosed with ESGD, were selected. Horses were confined to individual stalls, exercised once daily, and fed ad libitum hay, 1 kg of a low starch compound complementary feed and a mineral supplement. Half of the horses received a compound containing hydrolysed collagen (supplement) and the other half did not (control). At the start of the study, ESGD scores were 3.57 and 3.36 for the supplement and control group, respectively. After 4 weeks, the ESGD grades were significantly reduced in both groups (1.89 and 1.43, respectively, p < 0.01), and healing (ESGD < 2) occurred in 7 out of 14 horses. No treatment effect was observed (p = 0.75), and it was concluded that the change in husbandry overshadowed any potential effect of the compound. Severe ESGD can improve, and even heal, with the provision of a diet of ad libitum forage and a small amount of a compound complementary feed, without the use of omeprazole. A predictable daily routine, with a limited number of dedicated caretakers, may have contributed to the improvement of gastric health.

Methane production from Welsh ponies fed 2 isoenergetic diets (NE basis) at maintenance was studied in a crossover design with 4 mature geldings (230 ± 10.5 kg BW, mean ± SE). Treatments included a roughage-only (R) diet (5.1 kg DM/d) or a roughage plus cereal mix (RC) diet (2.5 kg DM hay/d plus 1.1 kg DM cereal mix/d). For both diets, the same grass hay was used (898 g DM/kg and 4.5 MJ NE_m/kg DM) and a commercial cereal mix was used in the RC diet (890 g DM/kg and 9.6 MJ NE_m/kg DM). Ponies were housed in pairs in climate-controlled respiration chambers. Carbon dioxide production (CO₂), oxygen (O₂) consumption, and CH₄ production were measured over 3 consecutive days. Heat production (HP) rates were calculated from gaseous exchange. Feces were collected quantitatively to determine dietary nutrient digestibility. Dry matter intake differed between diets (P < 0.0001), but NE intake was equal for both diets (22.3 ± 0.07 MJ NE_m/d). Organic matter digestibility was lower (P = 0.006) for the R diet (47.2%) than the RC diet (55.6%). Methane production was higher (P = 0.014) on the R diet (29.8 L·pony^–1·d^–1) compared to the RC diet (23.2 L·pony^–1·d^–1). Methane production expressed in liters/kilogram metabolic body weight (BW^0.75) per day tended (P = 0.064) to decrease with 21% for the RC group compared with the R group. Heat production, O₂ consumption, and CO₂ production were not affected by diet. Diurnal patterns of CH₄ production and HP were similar for both diets. Methane production increased slightly (P < 0.652) after feeding and was numerically lower for the RC diet for all time points throughout the day. For both diets, HP was higher after feeding than before feeding and decreased again within approximately 3 h after feeding. Isoenergetic replacement of roughage by a cereal mix reduces CH₄ production in ponies. No clear diurnal pattern in CH₄ emission can be discerned in ponies fed at maintenance. Copyright © 2015. American Society of Animal Science .

As colic and intestinal disorders are a major concern in horses, the aim of the present study was to investigate the effect of dietary supplementation of butyrate, known to have a diverse array of beneficial effects on intestinal health. The effect of micro-encapsulated sodium butyrate supplementation on gut histology and immunohistochemistry parameters was studied in 14 healthy warmblood horses destined for slaughter in two separate periods. Horses were fed a low fiber - high starch diet, designed to induce subsequent starch overflow in the large intestine, aiming to create a mild challenge for large intestinal health. Treatment included supplementation with either micro-encapsulated sodium butyrate (Excential Butycoat®, Orffa, Werkendam, the Netherlands) or placebo (containing only coating material). The horses were fed for 20 consecutive days at a dosage of 0.4 g/kg BW (body weight). At day 21, the horses were slaughtered and intestinal samples were collected for determination of gut pH, villus length, crypt depth and area % of CD3+ and CD20+ cells. Horses on the butyrate supplemented diet had significantly reduced crypt depths in the right dorsal colon compared to placebo-fed horses (P < 0.001). However, a treatment x period interaction (P = 0.002) was discovered regarding this parameter, which could not be explained by the authors. Further investigation into the number of KI67+ cells in the RDC crypts did not reveal any significant differences between treatments (P = 0.650), indicating that the reduction in crypt depth in butyrate-fed horses could not be explained by a significant difference in cellular proliferation. Intestinal pH, villus length and expression of intestinal CD3+ and CD20+ cells were not significantly affected by treatment at any intestinal level. Our data indicate that supplementation of micro-encapsulated sodium butyrate to the equine diet did not influence gut histology (with the exception of a decrease found in the crypts of the RDC) or immunohistochemistry parameters in healthy horses. Further research is warranted to investigate the impact of butyrate supplementation in horses with intestinal disease.