Multi-omics reveal the gut microbiota-mediated severe foraging environment adaption of small wild ruminants in the Three-River-Source National Park, China.

The Tibetan antelope (Pantholops hodgsonii), blue sheep (Pseudois nayaur), and Tibetan sheep (Ovis aries) are the dominant small ruminants in the Three-River-Source National Park (TRSNP). However, knowledge about the association between gut microbiota and host adaptability remains poorly understood. Herein, multi-omics sequencing approaches were employed to investigate the gut microbiota-mediated forage adaption in these ruminants. The results revealed that although wild ruminants (WR) of P. hodgsoni and P. nayaur were faced with severe foraging environments with significantly low vegetation coverage and nutrition, the apparent forage digestibility of dry matter, crude protein, and acid detergent fiber was significantly higher than that of O. aries. The 16s rRNA sequencing showed that the gut microbiota in WR underwent convergent evolution, and alpha diversity in these two groups was significantly higher than that in O. aries. Moreover, indicator species, including Bacteroidetes and Firmicutes, exhibited positive relationships with apparent forage digestibility, and their relative abundances were enriched in the gut of WR. Enterotype analysis further revealed that enterotype 1 belonged to WR, and the abundance of fatty acid synthesis metabolic pathway-related enzyme genes was significantly higher than enterotype 2, represented by O. aries. Besides, the metagenomic analysis identified 14 pathogenic bacterial species, among which 10 potentially pathogenic bacteria were significantly enriched in the gut microbiota of O. aries. Furthermore, the cellulolytic strains and genes encoding cellulase and hemicellulase were significantly enriched in WR. In conclusion, our results provide new evidence of gut microbiota to facilitate wildlife adaption in severe foraging environments of the TRSNP, China.

Targeted and untargeted metabolomics reveals meat quality in grazing yak during different phenology periods on the Qinghai-Tibetan Plateau.

Yak meat is more popular among consumers because of its high nutritional value, but little attention has been paid to its meat quality, which is affected by different phenology periods grass. We hypothesized that seasonal variations in grass composition influenced the ruminal bacteria community, and eventually affected the meat quality of yaks. This study aims to investigate the relationship of meat quality in grazing yak as well as the key rumen bacteria using targeted and untargeted metabolomics and 16S rRNA during different phenology periods. The main three altered metabolic pathways in grazing yak, including amino acids biosynthesis, glutathione metabolism, and fatty acids biosynthesis, were found in the grass period (GP) group compared to the regreen period (RP) and hay period (HP) groups. The GP group had higher concentrations of flavor amino acids (FAA), polyunsaturated fatty acids (PUFA), and a lower ratio of n-6/n-3 compared with the RP group. Correlation analysis results showed that Rikenellaceae_RC9_gut_group was positively correlated with fatty acids and lipid metabolites, which might be involved in lipid metabolism. Pediococcus had a positive correlation with biological peptides, which could be involved in the metabolism of bioactive compounds. In conclusion, grass in different phenology periods was associated with modified amino acids and fatty acids composition of yak meat as well as altered regulation of biological pathways, which was correlated with changes in rumen bacterial communities.

Effects of the alpine meadow in different phenological periods on rumen fermentation and gastrointestinal tract bacteria community in grazing yak on the Qinghai-Tibetan Plateau.

BACKGROUND: In this study, we investigated the effects of alpine meadow in different phenological periods on ruminal fermentation, serum biochemical indices, and gastrointestinal tract microbes in grazing yak on the Qinghai-Tibetan Plateau. A total of eighteen female freely grazing yaks with an average age of 3 years old and a body weight of 130 ± 19 kg were selected. According to the plant phenological periods, yaks were randomly allocated to one of three treatments: (1) regreen periods group (RP, n = 6); (2) grassy periods group (GP, n = 6); and (3) hay periods group (HP, n = 6). At the end of the experiment, the blood, rumen fluids, and rectal contents were collected to perform further analysis. RESULTS: The concentrations of total volatile fatty acid (TVFA), acetate, glucose (GLU), triglyceride (TG), cholesterol (CHO), high density lipoprotein (HDL), and low density lipoprotein (LDL) were higher in the GP group than in the HP group (P < 0.05). However, compared with the RP and GP groups, the HP group had higher concentrations of isobutyrate, isovalerate, valerate, and creatinine (CREA) (P < 0.05). The abundance of Prevotella in the rumen, and the abundances of Rikenellaceae_RC9_gut_group, Eubacterium_coprostanoligenes_group, and Prevotellaceae_UCG-004 in the gut were higher in the GP group compared with the HP group (P < 0.05). The HP had higher abundance of Eubacterium_coprostanoligenes_group in the rumen as well as the abundances of Romboutsia and Arthrobacter in the gut compared with the RP and GP groups (P < 0.05). CONCLUSIONS: Based on the results of rumen fermentation, serum biochemical, differential biomarkers, and function prediction, the carbohydrate digestion of grazing yak would be higher with the alpine meadow regreen and grassy due to the gastrointestinal tract microbes. However, the risk of microbe disorders and host inflammation in grazing yak were higher with the alpine meadow wither.

Evaluation of the Longissimus Thoracis et Lumborum Muscle Quality of Chaka and Tibetan Sheep and the Analysis of Possible Mechanisms Regulating Meat Quality.

This study aimed to comprehensively evaluate the characteristics in the longissimus thoracis et lumborum (LTL) muscle of Chaka (CK) sheep and Tibetan (TB) sheep, and transcriptomics-metabolomics association analysis was used to find the possible genes, differential metabolites, and significant differential metabolic pathways that lead to meat quality differences. Based on the researched results, the nutritional quality of meat, including the contents of ether extract (11.95% vs. 10.56%), unsaturated fatty acid (51.20% vs. 47.69%), and polyunsaturated fatty acid (5.71% vs. 3.97%), were better in TB sheep than in CK sheep, while the CK sheep has better muscle fiber characteristics, such as the total number (62 vs. 45) and muscle fiber density (1426.54 mm2 vs. 1158.77 mm2) and flavor. Omics research has shown that the key differential metabolites and metabolic pathways were dominated by amino acid metabolism, particularly the glutathione metabolism, taurine and hypotaurine metabolism, and lipid metabolism-related pathways, such as glycerophospholipid metabolism and the sphingolipid signaling pathway. The intermediate metabolite sn-Glycerol 3-phosphoethanolamine played a key role in determining sheep meat quality, which was regulated by GPAT2, PLPP2, AGPAT1, PNPLA2, and GPAT4 and correlated with meat color, texture, and flavor. Overall, these results will provide effective information and more evidence to support further exploration of valuable biomarkers of meat quality.

Rumen Microbiome Reveals the Differential Response of CO2 and CH4 Emissions of Yaks to Feeding Regimes on the Qinghai-Tibet Plateau.

Shifts in feeding regimes are important factors affecting greenhouse gas (GHG) emissions from livestock farming. However, the quantitative values and associated drivers of GHG emissions from yaks (Bos grunniens) following shifts in feeding regimes have yet to be fully described. In this study, we aimed to investigate CH4 and CO2 emissions differences of yaks under different feeding regimes and their potential microbial mechanisms. Using static breathing chamber and Picarro G2508 gas concentration analyzer, we measured the CO2 and CH4 emissions from yaks under traditional grazing (TG) and warm-grazing and cold-indoor feeding (WGCF) regimes. Microbial inventories from the ruminal fluid of the yaks were determined via Illumina 16S rRNA and ITS sequencing. Results showed that implementing the TG regime in yaks decreased their CO2 and CH4 emissions compared to the WGCF regime. The alpha diversity of ruminal archaeal community was higher in the TG regime than in the WGCF regime. The beta diversity showed that significant differences in the rumen microbial composition of the TG regime and the WGCF regime. Changes in the rumen microbiota of the yaks were driven by differences in dietary nutritional parameters. The relative abundances of the phyla Neocallimastigomycota and Euryarchaeota and the functional genera Prevotella, Ruminococcus, Orpinomyces, and Methanobrevibacter were significantly higher in the WGCF regime than in the TG regime. CO2 and CH4 emissions from yaks differed mainly because of the enrichment relationship of functional H2- and CO2-producing microorganisms, hydrogen-consuming microbiota, and hydrogenotrophic methanogenic microbiota. Our results provided a view that it is ecologically important to develop GHG emissions reduction strategies for yaks on the Qinghai-Tibet Plateau based on traditional grazing regime.

Comparative Analysis of the Composition of Fatty Acids and Metabolites between Black Tibetan and Chaka Sheep on the Qinghai-Tibet Plateau.

The objective of this study was to investigate and compare fatty acids and metabolites in the longissimus dorsi muscle between Black Tibetan and Chaka sheep grazing in a highly saline environment. A total of eight castrated sheep (14 months old) with similar body weights (25 ± 2.2 kg) were selected. The experimental treatments included Black Tibetan (BT) and Chaka sheep (CK) groups, and each group had four replications. The experiment lasted for 20 months. All sheep grazed in a highly saline environment for the whole experimental period and had free access to water. The results showed that the diameter (42.23 vs. 51.46 µm), perimeter (131.78 vs. 166.14 µm), and area of muscle fibers (1328.74 vs. 1998.64 µm2) were smaller in Chaka sheep than in Black Tibetan sheep. The ash content in the longissimus dorsi was lower in Chaka sheep than in Black Tibetan sheep (p = 0.010), and the contents of dry matter (DM), ether extract (EE), and crude protein (CP) in the longissimus dorsi showed no differences (p > 0.05). For fatty acids, the proportions of C10:0, C15:0, and tC18:1 in the longissimus dorsi were higher in Chaka sheep than in Black Tibetan sheep (p < 0.05). However, all other individual fatty acids were similar among treatments, including saturated fatty acids (SFAs), unsaturated fatty acids (UFAs), monounsaturated fatty acids (MUFAs), polyunsaturated fatty acids (PUFAs), and the ratios of n-6 PUFAs to n-3 PUFAs and PUFAs to SFAs (p > 0.05). A total of 65 biomarkers were identified between the two breeds of sheep. Among these metabolites, 40 metabolic biomarkers were upregulated in the CK group compared to the BT group, and 25 metabolites were downregulated. The main metabolites include 30 organic acids, 9 amino acids, 5 peptides, 4 amides, 3 adenosines, 2 amines, and other compounds. Based on KEGG analysis, eight pathways, namely, fatty acid biosynthesis, purine metabolism, the biosynthesis of unsaturated fatty acids, renin secretion, the regulation of lipolysis in adipocytes, neuroactive ligand−receptor interaction, the cGMP-PKG signaling pathway, and the cAMP signaling pathway, were identified as significantly different pathways. According to the results on fatty acids and metabolites, upregulated organic acid and fatty acid biosynthesis increased the meat quality of Chaka sheep.

Application of a Gas-Kinetic BGK Scheme in Thermal Protection System Analysis for Hypersonic Vehicles.

One major problem in the development of hypersonic vehicles is severe aerodynamic heating; thus, the implementation of a thermal protection system is required. A numerical investigation on the reduction of aerodynamic heating using different thermal protection systems is conducted using a novel gas-kinetic BGK scheme. This method adopts a different solution strategy from the conventional computational fluid dynamics technique, and has shown a lot of benefits in the simulation of hypersonic flows. To be specific, it is established based on solving the Boltzmann equation, and the obtained gas distribution function is used to reconstruct the macroscopic solution of the flow field. Within the finite volume framework, the present BGK scheme is specially designed for the evaluation of numerical fluxes across the cell interface. Two typical thermal protection systems are investigated by using spikes and opposing jets, separately. Both their effectiveness and mechanisms to protect the body surface from heating are analyzed. The predicted distributions of pressure and heat flux, and the unique flow characteristics brought by spikes of different shapes or opposing jets of different total pressure ratios all verify the reliability and accuracy of the BGK scheme in the thermal protection system analysis.

Changes in the Fermentation and Bacterial Community by Artificial Saliva pH in RUSITEC System.

The purpose of the study was to assess the artificial saliva (AS) pH on ruminal fermentation and rumen bacteria community in the rumen simulation technique (RUSITEC) system. The experiment was performed in two treatments (low AS pH vs. high AS pH) with four replicates. The low AS pH was sustained by altering the composition of the AS (NaHCO3 from 9.8 to 1.96 g/L, Na2HPO4 from 9.3 to 1.86 g/L) according to McDougall's method. The diets were supplemented with 16 g basic diets with forage to the concentrate ratio of 50:50. The experiments were conducted over 13-day incubation periods, with 9 days adaption and 4 days sample collection. The results showed low AS pH decreased dry matter (DM) degradability (64.37 vs. 58.67%), organic matter (OM) degradability (64.38 vs. 59.32%), neutral detergent fiber (NDF) degradability (46.87 vs. 39.94%), acid detergent fiber (ADF) degradability (38.16 vs. 31.13%), and crude protein (CP) degradability (70.33 vs. 62.99%), respectively. Compared with the high AS pH, the low AS pH increased the proportion of butyrate (P = 0.008) and decreased the proportion of propionate (P < 0.001). At the bacteria community, the low AS pH increased the abundances of Spirochaetes (P = 0.001) and Synergistetes (P = 0.004) and decreased the Verrucomicrobia abundance (P = 0.004) in solid-associated bacteria. At the genus level, the low AS pH increased the abundance of Lactobacillus (P = 0.050) and decreased the abundance of Schwartzia (P = 0.002) in solid-associated bacteria. The abundances of Prevotellaceae_YAB2003_group (P = 0.040), Schwartzia (P = 0.002), and Ruminobacter (P = 0.043) were lower in the low AS pH group compared with the high AS pH group in liquid-associated bacteria. Low AS pH decreased the number of Ruminococcus albus, Ruminococcus flavefaciens, Fibrobacter succinogenes (P < 0.001) both in the solid- and liquid-associated bacteria, respectively. The results of the present study included three groups of bacteria communities according to the different sensitives to rumen pH: the abundances of Lactobacillus, Succinivibrio, and Prevotella_7 are increased with decreasing AS pH; the amounts of R. albus, R. flavefaciens, F. succinogenes as well as the abundances of Schwartzia and Ruminobacter decreased with the reducing AS pH; the abundances of Selenomonas_1, Rikenellaceae_RC9_gut_group, and Succiniclasticum were not affected by the AS pH in RUSTITEC.

The Destruction of the Anaerobic Environment Caused by Rumen Fistula Surgery Leads to Differences in the Rumen Microbial Diversity and Function of Sheep.

This experiment was to study the impact of rumen fistula surgery on the rumen microbios in sheep. Six male adult Hu sheep (48.8 ± 0.23 kg, 0.5 years) were fed at 0700 and 1,800 with ad libitum access to water. The rumen fistula was installed in the same batch from 0600 to 0900. Monitoring the dry mater intake and the output of dry mater faces 1, 2, 4, 6, 8, 10, 12, 14 days after fistulated surgery. The collection of rumen fluid was arranged at 1d during rumen surgery (DRS1), 3d after rumen surgery (ARS3), and 14d after rumen surgery (ARS14) for volatile fatty acid (VFA) and DNA extraction for sequencing and bioinformatics analysis. There was no difference in DMI, the pH apparent digestibility of dry matter, organic matter, neutral detergent fiber, acid detergent fiber both before and 14 days after surgery. Increases were observed in the acetate and total VFA at ARS3. There was no difference in digestion of dry material, organic material, neutral detergent fiber, and acid detergent fiber before and after surgery. The relative abundance of Bacteroides decreased from 61.96% at DRS1 to 28.85% at ARS3. In comparison with the DRS1 and ARS3, the relative abundance of Firmicutes in the ARS14 increased to 44.58% (P < 0.01). Proteobacteria increased from 11.33% at DRS1 to 51.66% at ARS3 and then decreased to 11.39% at ARS14. Prevotella decreased form 61.06% at DRS1 to 28.04% in the ARS3. Succinivibrio increased from 8.32% at DRS1 to 48.58% at ARS3, but decreased to 10.43% in the ARS14. Compared with DRS1 and ARS3, the ARS14 was higher in the Simpson and Shannon index. As for the BugBase function prediction, rumen fistula surgery increased the microorganism abundance of aerobic and facultative anaerobic phenotype, and anaerobic phenotype was decreased in the ARS3. There was higher microorganism abundance of aerobic phenotype in the ARS14 than before fistula installation. In conclusion, the rumen fistula surgery destroys the anaerobic environment of rumen, leading to differences in rumen microbial diversity and function, but the apparent digestibility and total VFA were not affected.

Effects of supplementation of nonforage fiber source in diets with different starch levels on growth performance, rumen fermentation, nutrient digestion, and microbial flora of Hu lambs.

The objectives were to evaluate the effects of fiber source and dietary starch level on growth performance, nutrient digestion, rumen parameters, and rumen bacteria in fattening Hu lambs. A total of 360 Hu lambs (BW = 24.72 ± 0.14 kg, 2 months old) were subjected to a 2 × 3 factorial arrangement. Lambs randomly assigned 6 treatments with 6 repetitions (10 lambs per repetition) of each treatment. Six treatments were formulated to include the fiber sources with three starch levels. The experiment lasted a 63 d. The amount of feed, orts, and total feces were sampled on the 42nd day of the experiment. Rumen fluid samples were collected after 2 h of morning feeding on day 56. Rumen contents were collected last day after the selected lambs were slaughtered. Increasing the starch content decreased the digestibility of neutral detergent fiber (NDF, P = 0.005). Increasing the starch level increased the proportions of propionate (P = 0.002) and valerate (P = 0.001) and decreased the proportion of acetate (P < 0.001) and the ratio of acetate to propionate (P = 0.005). The abundance of Fibrobacter succinogenes was affected by an interaction between the fiber source and the starch level (P < 0.001). Fibrobacter succinogenes tended to be greater in lambs fed SH than in lambs fed BP (P = 0.091), which was greater in lambs fed high starch levels than in lambs fed low starch levels (P = 0.014). Increasing the starch level increased Streptococcus bovis abundance (P = 0.029) and decreased total bacteria (P = 0.025). At the genus level, increasing the starch level reduced the abundance of Butyrivibrio_2 (P = 0.020). Nevertheless, the final body weight (BW) and acid detergent fiber (ADF) digestibility were greater (P < 0.01) in lambs fed soybean hull (SH) than in lambs fed BP. The proportion of butyrate was greater (P = 0.005), while the rumen pH was lower (P = 0.001) in lambs fed beet pulp (BP) than in those fed SH. The abundances of Succiniclasticum, Candidatus_Saccharimonas, Ruminococcus_1, and Christensenellaceae_R-7 were greater in lambs fed SH than in those fed BP (P < 0.050), whereas the abundance of Fibrobacter was lower (P = 0.011). The predominant microbial phyla in all of the groups were Firmicutes, Bacteroidetes, and Fibrobacteres. Changing the starch level for fiber sources mainly changed the rumen community in terms of the phylum and genus abundances. Lambs fed SH with low starch level increased the final BW without affecting total volatile fatty acids (TVFA) concentrations.

Effects of Dietary Barley Starch Contents on the Performance, Nutrient Digestion, Rumen Fermentation, and Bacterial Community of Fattening Hu Sheep.

The objective of this experiment was to investigate the effects of substituting corn starch (CS) with barley starch (BS) on the growth performance, nutrient digestion, rumen fermentation, and bacterial community of fattening Hu sheep. Seventy-two Hu lambs with similar initial body weight (BW, 29.70 ± 1.70 kg) were randomly assigned to four treatments, with 18 lambs per group. The four experimental diets have identical starch contents but with different starch sources as 100% starch from corn (BS-0), 33% starch from barley and 67% starch from corn (BS-33), 67% starch form barley and 33% starch from corn (BS-67), and 100% starch from barley (BS-100). All lambs were reared in individual units and fed high-concentrate diets (85% concentrate in diets based on dry matter [DM]). The experimental period included 7 days for adaptation and 63 days for data collection. Sixteen ruminal cannulated Hu sheep were divided into 4 groups and received the four experimental diets to determine the dynamics of ruminal pH. The average daily gain (ADG), and BW gain of lambs linearly decreased (p < 0.05), whereas the feed to gain ratio linearly increased (p < 0.05) with increasing dietary proportions of BS. Digestibility of DM, organic matter, neutral detergent fiber, acid detergent fiber, starch, and gross energy (GE) decreased (p < 0.05) with increasing dietary BS contents. Ruminal mean pH decreased (p < 0.05) with increasing proportions of dietary BS, accompanied with linearly increased (p < 0.05) time and area of ruminal pH below 5.80 or 5.60. Increasing dietary proportions of BS linearly decreased (p < 0.05) the molar proportion of acetate, but linearly increased (p < 0.05) the molar proportion of propionate. Sheep of the BS-0 and BS-33 treatments had a less (p < 0.05) relative abundance of Selenomonas ruminantium than that of sheep of the BS-67 treatment, but a greater (p < 0.05) relative abundance of Ruminococcus albus than that of sheep of the BS-100 treatment (p < 0.05). In conclusion, feeding a high-concentrate corn-based diet for fattening Hu sheep improved the performance and rumen fermentation parameters when compared to the barley-based diet.