In the early 1990s, the equine hindgut microbial ecosystem looked like a “black box.” Its vital role in hydrolyzing and fermenting fiber, thus providing energy to the host, was recognized. Although there was a critical lack of information on the hindgut microbes, their role in preventing intestinal diseases was suggested. Traditionally, the microbes of the horse hindgut were studied using culture-dependent techniques. More recently, culture-independent methods have been used and provided further insight. This review presents the history and updated knowledge regarding the microbes that live inside the different intestinal ecosystems and which collective genomes compose the hindgut microbiome. In the first section, the quantification and diversity are described for each microbial community as well as the implication of plant fiber degradation and their crucial role for an herbivore host. The microbial communities are presented in chronological order of discovery: due to their large size, protozoa were brought to light as early as 1843 in the horse cecum; in 1897, bacteria were described in the horse intestine; as early as 1910, monoflagellated eukaryotic organisms resembling protozoa were observed in the horse cecum; since then, they have been identified to be zoospores of anaerobic fungi; in 1970, bacteriophage-like particles were recognized in the cecum and colon of pony and horse; and finally, in 1996, archaea were identified in the horse cecum. The second section discusses the variations that can occur between digestive segments or between individuals. The representativeness of the fecal microbiota to the hindgut one is debated, especially as the majority of recent studies conducted on the horse hindgut are in fact focused on the feces, rather than the cecum or colon. Also, the representation of microbiota between individuals is questioned. It has long been suggested in the literature that some ponies or horses that were more susceptible to intestinal diseases may harbor a specific intestinal microbiota. Alternatively, some new studies aim at identifying a core microbiome between all individual equine. A deeper knowledge of the microbiome and its core may allow improvement of nutrition and health, understanding of the onset of digestive diseases, and the development tools for health monitoring and disease prevention. Copyright © 2016. American Society of Animal Science .

In horses, abrupt changes from high-fiber (HF) to high-starch (HS) diets can affect the cecal and colonic microbiota. This study investigated modifications and recovery of fecal microbiota after two consecutive abrupt dietary changes. Twelve horses fed HF for 2 weeks were changed to HS for 5 days then returned to HF for 7 weeks. Six received lactic acid bacteria supplementation. Bacterial population diversity, structure, and activity, especially fibrolysis, were assessed to obtain an overview of alteration in hindgut microbiota. Two days after the abrupt change from HF to HS, the findings in feces were consistent with those previously reported in the cecum and colon, with a decrease in fibrolytic activity and an increase in amylolytic activity. Fecal parameters stabilized at their basal level 3–4 weeks after the return to HF. A bloom of cellulolytic bacteria and lower pH were observed after 1.5 weeks, suggesting a higher level of fiber degradation. In supplemented horses the relative abundance of potentially fibrolytic genera was enhanced 2 days after HS and 2 days to 2–3 weeks after the return to HF. Fecal analysis could be a promising technique for monitoring hindgut microbial variations accompanying dietary changes.

The equine hindgut ecosystem is specialized in dietary fibers’ fermentation to provide horses’ energy and contribute to its health. Nevertheless, antibiotics are known to disrupt the hindgut microbiota, affecting the fibrolytic activity of bacteria and the intestinal immune balance, leading to diseases. This in vivo study used a general and comprehensive approach for characterizing the hindgut ecosystem of 9 healthy horses over 28 days in response to a 5-day challenge with oral trimethoprim-sulfadiazine (TMS), with a special emphasis on microbial fibrolytic activity and the host immune response. Horses were supplemented with two doses of Lactobacillus acidophilus, Ligilactobacillus salivarius (formerly L. salivarius), and Bifidobacterium lactis blend or a placebo in a 3 × 3 Latin square design. Changes in fecal microbiota were investigated using 16S rRNA sequencing. Clostridioides difficile was quantified in feces using quantitative polymerase chain reaction. Anaerobic microbiological culture was used to enumerate functional bacterial groups (cellulolytic, amylolytic, and lactic acid-utilizing). The environmental dimensions were assessed by measuring the concentrations of volatile fatty acids (VFAs) and lactic acid using biochemical methods, and changes in pH and dry matter weight. Systemic and local inflammation was evaluated by determination of cytokine and immunoglobulin (Ig)A concentrations in the serum and secretory IgA (SIgA) concentrations in the feces using immuno-enzymatic methods. Oral TMS treatment strongly altered the whole hindgut ecosystem by 2 days after the first administration. Bacterial diversity decreased in proportion to the relative abundance of fibrolytic genera, which coincided with the decrease in the concentration of cellulolytic bacteria. At the same time, the composition of microbiota members was reorganized in terms of relative abundances, probably to support the alteration in fibrolysis. C. difficile DNA was not found in these horses, but the relative abundances of several potential pathobiont genera increased. 2 days after the first TMS administration, fecal concentrations of VFAs and SIgA increased in parallel with fecal water content, suggesting an alteration of the integrity of the hindgut mucosa. Recovery in bacterial composition, functions, and immune biomarkers took 2–9 days after the end of TMS administration. Supplementation with this bacterial blend did not limit bacterial alteration but might have interesting mucosal immunomodulatory effects.

Fibrous feeds are essential for horses. When developing feeding regimens promoting health and performance, we need to understand the digestion of plant cell walls and the functioning of the hindgut microbial ecosystem. Our objective was to investigate the effect of grass fibre maturity and legume forage on the hindgut microbiota and its activity. Six caecum and colon fistulated geldings were fed three diets differing in fibre composition: concentrate and late harvested grass haylage (35:65 energy ratio) (C); early and late harvested grass haylage (80:20) (G); lucerne and late harvested grass haylage (80:20) (L) for 28 days in a Latin-square design. No differences were measured in total bacteria concentrations, fungi and protozoa numbers nor in cellulolytic bacteria concentrations between the diets. Short-chain fatty acid concentrations did not differ between diets, but a lower (acetate + butyrate)/propionate ratio when the horses were fed Diet C, compared to G and L, was observed, suggesting lower fibrolytic and higher amylolytic activity. The pH increased when the horses were fed Diet L and decreased when fed C and G from caecum to faeces. The buffering capacity (BC) of hindgut digesta was five to fifteen-fold higher than that of the feeds, suggesting a decreased effect of feed BC as digesta travelled through the digestive tract. In conclusion, an early harvested forage opens up the possibility for forage-only diets, providing high energy without the negative effects of concentrate.