The intestinal microbiota plays an important role in the health and metabolism of the host. Next‐generation sequencing technology has enabled the characterization of the gut microbiota of several animal species. We analyzed the intestinal microbiota in six different parts of the gastrointestinal tracts (GITs) of five Mongolian horses by sequencing the 16S rRNA gene V3‐V4 hypervariable region. All horses were kept in the natural habitat of the Inner Mongolia grassland. Significant differences were observed among the microbiota compositions of the distinct GIT regions. In addition, while the microbial community structures of the small and large intestine were significantly different, those of the cecum and colon were similar. In the foregut, Firmicutes (65%) and Proteobacteria (23%) were the most abundant, while Firmicutes (45%) and Bacteroidetes (42%) were the most common in the hindgut. At the level of family, Ruminococcaceae (p = .203), Lachnospiraceae (p = .157), Rikenellaceae (p = .122), and Prevotellaceae (p = .068) were predominant in the hindgut, while the relative abundance of the Akkermansia genus (5.7%, p = .039) was higher in the ventral colon. In terms of the putative functions, the ratio of microbial abundance in the different parts of the GIT was similar, the result can help characterize the gut microbial structure of different animals.

To investigate the effects of calcium propionate (CaP) supplementation on performance, the development of the internal organ, and gastrointestinal tract of calves at various stages of growth period, 54 male Jersey calves (age = 7 ± 1 d, body weight(BW) = 23.1 ± 1.2 kg) were randomly allocated to three treatment groups. While control calves were fed basis dietary with no additives (0CaP), other treatment calves were fed basis dietary supplementation with CaP at 50 (5CaP) or 100 (10CaP) g kg⁻¹ dry matter. The experiment lasted 160 d and was divided into three feeding stages: Stage 1 (d 0 to 30), Stage 2 (d 31 to 90), and Stage 3 (d 91 to160). Six calves from each group were randomly selected and slaughtered on days 30, 90, and 160 when at the conclusion of each experimental feeding stage. The BW of calves increased with 10CaP after feeding 90 d, whereas it increased with 5CaP and 10CaP at feeding 120d and 160d compared to 0CaP. The 10CaP group improved average daily gain (ADG) of calves at stage 2, and d120-160 of stage 3 compared with the 0CaP group. The ADG of 5CaP was greater than the 0CaP group only at 120–160 d of stage 3 compared with the 0CaP group. The results of feed efficiency were in agreed with ADG as no dry matter intake difference at all stages of growth period. The 10CaP treatment exhibited the greatest spleen weight among the treatment at the end of the experiment; the liver weight of the 5CaP and 10CaP calves at feeding 90 d and of the 10CaP calves at feeding 160 d and were greater than those of the 0CaP animals. The CaP at the tested doses increased the rumen weight after feeding 90d of Jersey calves, and also improved the development of intestine. In conclusion, dietary supplementation with calcium propionate at the tested doses caused a beneficial effect in the growth performance and gastrointestinal tract traits of Jersey calves, thus to add 10% CaP before feeding 90 days was better and 5% CaP supplementation was expected at the period for feeding 90 to160 d.

In response to muscle injury, muscle stem cells are stimulated by environmental signals to integrate into damaged tissue to mediate regeneration. L-leucine (L-leu), a branched-chain amino acid (BCAA) that belongs to the essential amino acids (AAs) of the animal, has gained global interest on account of its muscle-building and regenerating effects. The present study was designed to investigate the impact of L-leu exposure to promote the proliferation of equine skeletal muscle satellite cells (SCs) on the regulation of RNA networks, including mRNA, long non-coding RNA (lncRNA), covalently closed circular RNA (circRNA), and microRNA (miRNA) in skeletal muscles. Equine SCs were used as a cell model and cultured in different concentrations of L-leu medium. The cell proliferation assay found that the optimal concentration of L-leu was 2 mM, so we selected cells cultured with L-leu concentrations of 0 mM and 2 mM for whole-transcriptiome sequencing, respectively. By high-throughput sequencing analysis, 2470 differentially expressed mRNAs (dif-mRNAs), 363 differentially expressed lncRNAs (dif-lncRNAs), 634 differentially expressed circRNAs (dif-circRNAs), and 49 differentially expressed miRNAs (dif-miRNAs) were significantly altered in equine SCs treated with L-leu. To identify the function of autoimmunity and anti-inflammatory responses after L-leu exposure, enrichment analysis was conducted on those differentially expressed genes (DEGs) related to lncRNA, circRNA, and miRNA. The hub genes were selected from PPI Network, including ACACB, HMGCR, IDI1, HAO1, SHMT2, PSPH, PSAT1, ASS1, PHGDH, MTHFD2, and DPYD, and were further identified as candidate biomarkers to regulate the L-leu-induced proliferation of equine SCs. The up-regulated novel 699_star, down-regulated novel 170_star, and novel 360_mature were significantly involved in the competing endogenous RNA (ceRNA) complex network. The hub genes involved in cell metabolism and dif-miRNAs may play fundamental roles in the L-leu-induced proliferation of equine SCs. Our findings suggested that the potential network regulation of miRNAs, circ-RNAs, lncRNAs, and mRNAs plays an important role in the proliferation of equine SCs, so as to build up new perspectives on improving equine performance and treatment strategies for the muscle injuries of horses.

This study was conducted to examine effects of nitrate on ruminal methane production, methanogen abundance, and composition. Six rumen-fistulated Limousin×Jinnan steers were fed diets supplemented with either 0% (0NR), 1% (1NR), or 2% (2NR) nitrate (dry matter basis) regimens in succession. Rumen fluid was taken after two-week adaptation for evaluation of in vitro methane production, methanogen abundance, and composition measurements. Results showed that nitrate significantly decreased in vitro ruminal methane production at 6 h, 12 h, and 24 h (P < 0.01; P < 0.01; P = 0.01). The 1NR and 2NR regimens numerically reduced the methanogen population by 4.47% and 25.82% respectively. However, there was no significant difference observed between treatments. The alpha and beta diversity of the methanogen community was not significantly changed by nitrate either. However, the relative abundance of the methanogen genera was greatly changed. Methanosphaera (PL = 0.0033) and Methanimicrococcus (PL = 0.0113) abundance increased linearly commensurate with increasing nitration levels, while Methanoplanus abundance was significantly decreased (PL = 0.0013). The population of Methanoculleus, the least frequently identified genus in this study, exhibited quadratic growth from 0% to 2% when nitrate was added (PQ = 0.0140). Correlation analysis found that methane reduction was significantly related to Methanobrevibacter and Methanoplanus abundance, and negatively correlated with Methanosphaera and Methanimicrococcus abundance.

Myostatin (MSTN), a member of the transforming growth factor-β superfamily, inhibits the activation of muscle satellite cells. However, the role and regulatory network of MSTN in equine muscle cells are not well understood yet. We discovered that MSTN knockdown significantly reduces the proliferation rate of equine muscle satellite cells. In addition, after the RNA sequencing of equine satellite cells transfected with MSTN-interference plasmid and control plasmid, an analysis of the differentially expressed genes was carried out. It was revealed that MSTN regulatory networks mainly involve genes related to muscle function and cell-cycle regulation, and signaling pathways, such as Notch, MAPK, and WNT. Subsequent real-time PCR in equine satellite cells and immunohistochemistry on newborn and adult muscle also verified the MSTN regulatory network found in RNA sequencing analysis. The results of this study provide new insight into the regulatory mechanism of equine MSTN.

The traditional horse industry has undergone a remarkable evolution, with horse racing emerging as a prominent and pivotal economic driver within the sector. Among the various breeds, Mongolian horses, renowned for their exceptional endurance and speed, occupy a significant position in the horse industry. To investigate their homeostasis mechanisms during and after a 20 km endurance exercise and identify novel oxidative-imbalance markers, we selected 12 two-year-old horses and collected blood samples at various time points before, during (at 5, 10, 15, and 20 km), and after the exercise (at 1, 2, 4, and 6 h post-exercise). These samples were analyzed for haematology, blood biochemistry, antioxidant enzyme activities, and liquid chromatography–mass spectrometry (LC-MS) metabolomics. Our results revealed significant changes in heart rate, speed, blood cells, and biochemical markers throughout the exercise. Antioxidant indicators decreased, while malondialdehyde increased, indicating oxidative imbalance post-exercise. Metabolomics analysis identified 122 differential metabolites, including uric acid and L-tyrosine, which were enriched in pathways related to energy metabolism. Uric acid and tyrosine correlated positively with serum creatine kinase, suggesting their potential as markers of oxidative-imbalance injury. These findings elucidate the mechanisms of endurance adaptability in Mongolian horses and provide a theoretical basis for mitigating oxidative imbalance, enhancing horse performance, and promoting the sustainable development of the equine industry.