Biomarkers for monitoring the equine large intestinal inflammatory response to stress-induced dysbiosis and probiotic supplementation.

Large intestine barrier disturbances can have serious consequences for the health of horses. The loss of mucosal integrity that leads to increased intestinal permeability may result from a local inflammatory immune response following alterations of the microbiota, known as dysbiosis. Therefore, our research aimed to identify noninvasive biomarkers for studying the intestinal permeability and the local inflammatory immune response in horses. Regarding the biomarkers used in other mammalian species, we measured the concentrations of lipopolysaccharides (LPS), reflected by 3-OH C14, C16, and C18 fatty acids, in blood, and fecal secretory immunoglobulin-A (SIgA). These biomarkers were evaluated in two trials including 9 and 12 healthy horses, which developed large intestinal dysbiosis experimentally induced by 5 d of antibiotic administration (trimethoprim sulfadiazine [TMS]) or 5 d of abrupt introduction of high starch levels (barley) into the diet. Horses were either control or supplemented with Lactobacillus acidophilus, Ligilactobacillus salivarius, and Bifidobacterium lactis. Correlations were performed between biomarkers and fecal bacterial diversity, composition, and function. No significant interaction between day and supplementation, or supplementation effect were observed for each biomarker. However, with the dietary stressor, a significant increase in blood concentrations of 3-OH C16 (P = 0.0125) and C14 (P = 0.0252) fatty acids was measured 2 d after the cessation of barley administration. Furthermore, with the antibiotic stressor, blood levels of 3-OH C16 progressively increased (P = 0.0114) from the first day to 2 d after the end of TMS administration. No significant day effect was observed for fecal SIgA concentrations for both stressors. These results indicate that both antibiotic- and diet-induced dysbiosis resulted in a local translocation of LPS 2 d after the cessation of the stressor treatments, suggesting an impairment of intestinal permeability, without detectable local inflammation. Blood LPS and fecal SIgA concentrations were significantly correlated with several bacterial variations in the large intestine, which are features of antibiotic- and diet-induced dysbiosis. These findings support the hypothesis that a relationship exists between dysbiosis and the loss of mucosal integrity in the large intestine of horses.

Horses can suffer from intestinal barrier disruption leading to permeability associated with local inflammation, which can result in discomfort and even disease. Intestinal barrier disruption may be a consequence of microbiota disturbances in the large intestine. Therefore, this study investigated the use of blood and fecal biomarkers for noninvasively assessing intestinal barrier permeability and inflammatory responses to microbial alterations. Two biomarkers were evaluated in healthy horses that were subjected to antibiotic- and diet-induced large intestine bacterial disturbances. Notably, the blood levels of the biomarkers increased 2 d after the cessation of both treatments, reflecting an abnormal intestinal barrier permeability. By contrast, the levels of fecal biomarker detected did not indicate the presence of inflammation. However, levels of the two biomarkers were significantly correlated with several bacterial variations in the feces, supporting the hypothesis that a relationship exists between microbiota disturbances and intestinal barrier disruption in the large intestine of horses.

Dietary composition and yeast/microalgae combination supplementation modulate the microbial ecosystem in the caecum, colon and faeces of horses.

Starchy diets can induce hindgut dysbiosis in horses. The present study evaluated the impact of a yeast (Saccharomyces cerevisiae) and microalgae (Aurantiochytrium limacinum) supplementation on caecal, colonic and faecal microbial ecosystem and on blood inflammatory parameters of horses fed high-fibre or high-starch diets. Six fistulated geldings in a 2 × 2 Latin-square design were alternatively supplemented and received during each period 100 % hay (4 weeks) followed by a 56/44 hay/barley diet (3 weeks). Caecal, colonic and faecal samples were collected 4 h after the morning meal three times per diet, at 5-d intervals, to measure bacterial composition and microbial end products. Blood was simultaneously collected for measuring inflammatory markers. The starchy diet clearly modified the microbial ecosystem in the three digestive segments, with an increase of the amylolytic function and a decrease of the fibrolytic one. However, no effect of the diet was observed on the blood parameters. When horses were supplemented, no significant change was found in lipopolysaccharides, PG-E2, serum amyloid A concentrations and complete blood count neither in cellulose-utilising, starch-utilising and lactate-utilising bacteria concentrations nor in the volatile fatty acids and lactate concentrations and pH. Under supplementation, relative abundance of Family XIII Clostridiales increased in caecum and faeces irrespective of diet and relative abundance of Veillonellaceae was higher during the hay/barley diet in colon and faeces. Most variations of faecal bacterial taxa under supplementation were not observed in the hindgut. However, all variations suggested that supplementation could increase fibrolytic function whatever the diet and limit dysbiosis when the horses' diet changed from high fibre to high starch.

Bacterial carbohydrate-degrading capacity in foal faeces: changes from birth to pre-weaning and the impact of maternal supplementation with fermented feed products.

The present study aimed at (1) describing age-related changes in faecal bacterial functional groups involved in carbohydrate degradation and in their activities in foals (n 10) from birth (day (d) 0) to 6 months (d180) and (2) investigating the effect of maternal supplementation (five mares per treatment) from d - 45 to d60 with fermented feed products on response trends over time of the foal bacterial carbohydratedegrading capacity. Maternal supplementation with fermented feed products stimulated foal growth from d0 to d60 and had an impact on the establishment of some digestive bacterial groups and their activities in foals from d0 to d5 but not in the longer term. Irrespective of the maternal treatment, total bacteria, total anaerobic, lactate-utilising and amylolytic bacteria were established immediately after birth (P<0·05) and were active as shown by the significant increase in total volatile fatty acids. In the foals of supplemented mares, total anaerobes and lactate utilisers were established rapidly between d0 and d2 (P=0·021 and 0·066, respectively) and the increase in the percentage of propionate occurred earlier (P=0·013). Maternal supplementation had no effect on the establishment of fibrolytic bacteria and their activity. Cellulolytic bacteria and Fibrobacter succinogenes first appeared at d2 and d5, and increased progressively, reaching stable values at d30 and d60, respectively. From the second week of life, the increase in the molar percentage of acetate and the ratio (acetate + butyrate):propionate (P<0·05) suggested that fibrolytic activity had begun. From d60, only minor changes in bacterial composition and activities occurred, showing that the bacterial carbohydrate-degrading capacity was established at 2 months of age.