Do microbial-gut-muscle mediated by SCFAs, microbial-gut-brain axis mediated by insulin simultaneously regulate yak IMF deposition?

Ruminant rumen plays an important role in the digestibility of cellulose, hemicellulose, starch and fat. In this study, the yaks under graze and stall feeding were chosen as the models of different rumen bacteria and intramuscular fat (IMF). The characteristics of IMF deposition, serum indexes in yaks were detected; the bacteria, metabolites in rumen was explored by 16S rRNA sequencing technology, untargeted metabolomics based on liquid chromatography-mass spectrometer and gas chromatography, respectively; the transcriptome of longissimus thoracis was identified by RNA-Sequencing analysis. Based on above results, a hypothesis that yak IMF deposition is regulated by the combined action of microbiome-gut-brain and muscle axis was proposed. The short-chain fatty acids (SCFAs) and neurotransmitters precursors like acetylcholine produced in yak rumen promoted insulin secretion via central nervous system. These insulin resulted in the high expression of SREBF1 gene by gut-brain axis; SCFAs can directly arrive to muscular tissue via blood circulation system, then activated the expression of PPARγ gene by gut-muscle axis. The expression of lipogenesis gene SCD, FABP3, CPT1, FASN and ACC2 was accordingly up-regulated. This study firstly introduce the theory of microbiome-gut-brain/muscle axis into the study of ruminant, and comprehensively expounded the regulatory mechanism of yak IMF deposition.

Effects of altitude and varieties on overwintering adaptability and cold resistance mechanism of alfalfa roots in the Qinghai-Tibet Plateau.

BACKGROUND: The roots are the main functional organs involved in the overwintering adaptability of alfalfa (Medicago sativa). However, it is still unclear how the roots are involved in the cold resistance in the high-altitude area of the Qinghai-Tibet Plateau (QTP). In this study, three winter-surviving 2-year-old alfalfa varieties (M. sativa Zhongmeng No.1, M. sativa Chiza No.1, and M. sativa Gongnong No.1) planted at two different altitudes (2812 m and 3109 m) in the northeast edge of the QTP were used to explore the cold-resistance mechanism. RESULTS: At low altitudes (2812 m), the overwintering rate, taproot length, root area, root surface area, and root average diameter, plant height, fresh yield and hay yield of M. sativa Zhongmeng No.1 were significantly higher (P < 0.01) than for the other two varieties. At high altitude (3109 m), lateral root length, number of lateral roots, main root dry weight, and lateral root dry weight of M. sativa Chiza No.1 were higher (P < 0.01) than the other two varieties. At low and high altitudes, the activities of peroxidase and catalase were higher (P < 0.05) in M. sativa Chiza No.1 during post-winter and pre-winter respectively. At low altitude, higher soluble sugar (P < 0.05) and proline (P < 0.01) contents were recorded during the pre- and post-winter periods. Membership function analysis showed that M. sativa Zhongmeng No.1 has the strongest cold resistance. The structural equation model showed that the overwintering rate of alfalfa was mainly affected by the morphological characteristics of roots and the physiological characteristics of roots, with contribution rates of 0.54 and 0.75 respectively, and the physiological characteristics of roots had the greatest effect on the overwintering rate. CONCLUSIONS: This study is of great significance to effectively solve the overwintering of alfalfa, the lack of high-quality legume forage resources, and promote the development of animal husbandry in the alpine areas of the QTP. © 2022 Society of Chemical Industry.

An increase in livestock density increases forage nutritional value but decreases net primary production and annual forage nutritional yield in the alpine grassland of the Qinghai-Tibetan Plateau.

Pasture biomass and quality are dependent on herbivore grazing and precipitation, but the responses of vegetation to the interactive effects of climate and grazing regimes remain unclear. We conducted an eight-year sheep grazing experiment with 4 stocking rates (0, 3.5, 5.5, and 7.5 sheep/ha) in an alpine meadow of the northeastern Tibetan Plateau. The above-ground net primary productivity (ANPP) and forage nutritional value (FNV) of four dominant species (Poa annua, Kobresia humilis, Astragalus adsurgens and Potentilla fruticosa) were measured during a wet year (360 mm rainfall) and a drought year (216 mm rainfall). The FNV was used as indicator of forage quality and was calculated from the crude protein (CP) content, in vitro true dry matter digestibility (IVTD), metabolic energy (ME) yield, and neutral detergent fiber (NDF) content of the plant. The stocking rate explained a minimum of 76% of the variations of ANPP, and the precipitation sub-additive effect for ANPP ranged from 5% to 12%. The interaction of sheep stocking rate and precipitation affected ANPP of the 4 species, except for P. fruticosa. The FNV of the pasture increased with increasing grazing pressure, but ANPP and forage nutritional yield (FNY) decreased. In calculating FNY, the increase in FNV did not compensate for the decrease in ANPP. In non-grazed plots, the CP yield declined sharply (18%-55%) in response to drought, but there was no effect on ME yield. The interaction between stocking rate and precipitation affected forage quality of the 4 plant species differently. The grassland ANPP and FNY could be maintained at a grazing intensity of 3.5 sheep/ha in wet and dry years. Our results highlight that stocking density affects pasture ANPP and FNV, and is contingent on rainfall.

Physiological adaptability of three gramineae plants under various vegetation restoration models in mining area of Qinghai-Tibet Plateau.

Large-scale coal mining activities on the QTP have affected the natural grassland ecosystem which will take decades to recover. Therefore, looking for an effective vegetation restoration method is the most effective way to restore the ecosystem. In this study, we explored the effects of three vegetation restoration modes (coal mine spoils planting (CK), organic fertilizer planting (OF), and cover soil planting (CS)) on the five cold-resistance related physiological indices of three native Gramineae plants (Elymus nutans, Poa crymophila and Festuca sinensis) growing in three different habitats (shady slope, sunny slope, and flat slope). The higher contents of soluble sugar (SS) in the Elymus nutans (213.46 mg/g) and Festuca sinensis (202.84 mg/g) were recorded under the CS treatment in the sunny slope habitat. The FP contents of Elymus nutans (421.31 µg/g), Poa crymophila (310.06 µg/g), and Festuca sinens (288.45 µg/g) were higher under the CK treatment in the flat ground habitat. The higher (P < 0.05) content of chlorophyll (Chl) was recorded in the Elymus nutans (2.13 mg/g) under the CS treatment in the sunny slope habitat compared to the other two plants. We observed lower MDA contents in the Elymus nutans (3.19 nmol/g) and Festuca sinensis (3.56 nmol/g) under the CK treatment growing in the shady slope habitat. In the shady slope habitat, the H2O2 contents of Elymus nutans (15.86 µmol/g), Poa crymophila (6.78 µmol/g), and Festuca sinensis (8.76 µmol/g) under the CS treatment were significantly lower compared to other treatments. Based on membership function method, the low-temperature adaptability order of the three restoration plants from strong to weak was Elymus nutans > Festuca sinensis > Poa crymophila. While the effectiveness order of the three vegetation restoration models in alpine mining areas from good to bad was CS > OF > CK. This study will provide a theoretical and guiding significance to evaluate the effectiveness of different restoration methods, formulate optimal restoration strategies and guide the sustainable restoration of the damaged ecosystems of the alpine mining areas.

Microbiome and Metabolomics Reveal the Effects of Different Feeding Systems on the Growth and Ruminal Development of Yaks.

The change in the feeding system can greatly improve the growth performance of the yak (Bos grunniens), an important livestock species in the plateau region. Here, we comprehensively compared the effects of different feeding systems on the growth performance and ruminal development of yaks, and investigated the effects of ruminal microorganisms and metabolites using the 16S rRNA gene sequencing and liquid chromatograph-mass spectrometer (LC-MS) technologies. We found that compared to traditional grazing feeding, house feeding significantly improved the growth performance (such as average daily gain and net meat weight) and rumen development of the yaks. At the genus level, the abundance of Rikenellaceae RC9 Gut group, Christensenellaceae R-7 group, Lachnospiraceae NK3A20 group, Ruminococcaceae UCG-014, and Prevotellaceae UCG-003 showed significant differences and was closely related to rumen development in the two distinct feeding systems. Also, metabolomics revealed that the change in the feeding system significantly affected the concentration and metabolic pathways of the related rumen metabolites. The metabolites with significant differences were significantly enriched in purine metabolism (xanthine, adenine, inosine, etc.), tyrosine metabolism (L-tyrosine, dopaquinone, etc.), phenylalanine metabolism (dihydro-3-caumaric acid, hippuric acid, etc.), and cAMP signaling pathway [acetylcholine, (-)-epinephrine, etc.]. This study scientifically support the house fattening feeding system for yaks. Also, our results provide new insights into the composition and function of microbial communities that promote ruminal development and in general growth of the yaks.

Transcriptome Analysis Reveals the Potential Role of Long Non-coding RNAs in Mammary Gland of Yak During Lactation and Dry Period.

The mammary gland is a remarkably dynamic organ of milk synthesis and secretion, and it experiences drastic structural and metabolic changes during the transition from dry periods to lactation, which involves the expression and regulation of numerous genes and regulatory factors. Long non-coding RNA (lncRNA) has considered as a novel type of regulatory factors involved in a variety of biological processes. However, their role in the lactation cycle of yak is still poorly understood. To reveal the involved mechanism, Ribo-zero RNA sequencing was employed to profile the lncRNA transcriptome in mammary tissue samples from yak at two physiological stages, namely lactation (LP) and dry period (DP). Notably, 1,599 lncRNA transcripts were identified through four rigorous steps and filtered through protein-coding ability. A total of 59 lncRNAs showed significantly different expression between two stages. Accordingly, the results of qRT-PCR were consistent with that of the transcriptome data. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses indicated that target genes of differentially expressed lncRNAs (DELs) were involved in pathways related to lactation, such as ECM-receptor interaction, PI3K-Akt signaling pathway, biosynthesis of amino acids and focal adhesion etc. Finally, we constructed a lncRNA-gene regulatory network containing some well known candidate genes for milk yield and quality traits. This is the first study to demonstrate a global profile of lncRNA expression in the mammary gland of yak. These results contribute to a valuable resource for future genetic and molecular studies on improving milk yield and quality, and help us to gain a better understanding of the molecular mechanisms underlying lactogenesis and mammary gland development of yak.

Plant communities responding to grazing pressure by sheep in an Alpine meadow.

The Chinese grassland ecosystem is an important national asset that not only impacts climate regulation, soil and water conservation, wind protection, and soil carbon and nitrogen fixation but is also an important contributor to maintaining grassland biodiversity while supporting livestock production. Grasslands are a key component contributing to the productivity of grazing animals but also provide basic food production via livestock grazing for herder survival. Grazing is the most basic means of grassland utilization but is considered one of the more important disturbance factors controllable by humans that has a universal and profound impact on the grassland ecosystem due to animal density and over grazing. For Alpine grasslands, it is not clear what grazing intensity (GI) can be achieved to improve plant biodiversity and vegetative nutritional value while improving sheep productivity. This field experiment was conducted for 7 yr comparing the impact of different GI on vegetation community characteristics, nutritional value, and sheep growth performance on the Alpine meadows of the Qinghai-Tibetan Plateau. The GI measured were: Control: 0 sheep/ha; Low: 3.7 sheep/ha; Medium: 5.3 sheep/ha; and Heavy: 7.6 sheep/ha. The grazing experiment started in 2008, but experimental data collection and analyses were collected for the final 4 yr of 2015 through 2018. All grazing intensities >0 sheep/ha reduced (P < 0.05) plant height (27%, 46%, and 48%, respectively, for 3.7, 5.3, and 7.6 sheep/ha), ground coverage (16%, 24%, and 48%), and above ground biomass (2%, 42% and 53%) of the various plant communities while increasing (P < 0.05) the grass community density (individuals/m2) compared to a nongrazed Control. With increasing GI, the community height, coverage, and above-ground biomass decreased (P < 0.05), and the plant community density increased then decreased (P < 0.05) compared to Control. As GI increased, the available community biomass nutritional quality increased (P < 0.05). Comprehensive analysis showed that the community density (quantity) and nutritional quality were the highest when the GI was 5.3 sheep/ha. The higher the GI, the greater the grass's nutritive value with lower above-ground net primary production (ANPP). When GI was the highest, the average daily gain (ADG) per hectare was the highest in the short term, but the highest GI endangers the ANPP and profitability of the grassland grazing ecosystem in the long term. Targeting a moderate GI (5.3 sheep/ha) can provide 78% of the ADG per hectare of the highest GI, which meets the requirement of maintaining a sustainable grazing grassland.

The Study of the Response of Fat Metabolism to Long-Term Energy Stress Based on Serum, Fatty Acid and Transcriptome Profiles in Yaks.

Long-term energy stress (ES) during the cold season is a serious problem for the breeding of yaks. In this paper, the response of fat metabolism in yaks to long-term ES during the cold season was studied. Gas chromatography (GC) analysis showed that the percentage of saturated fatty acids (SFAs) in the subcutaneous fat of the yaks in the ES group was 42.7%, which was less than the 56.6% in the CO group (p < 0.01) and the percentage of polyunsaturated unsaturated fatty acids (PUFAs) in the subcutaneous fat of the yaks in the ES group was 38.3%, which was more than the 26.0% in the CO group (p < 0.01). The serum analysis showed that fatty acid oxidation in yaks was increased under long-term ES. In the subcutaneous fat of yaks under long-term ES, the gene expression levels of glycerol-3-phosphate acyltransferase 4 (GPAT4), hormone-sensitive lipase (HSL), patatin-like phospholipase domain-containing protein 2 (PNPLA2), acyl-CoA dehydrogenase (ACAD), acyl-coenzyme A thioesterase 8 (ACOT8), facilitated glucose transporter (GLUT4), 3-oxoacyl-[acyl-carrier-protein] synthase (OXSM), oestradiol 17-beta-dehydrogenase 8 (HSD17B8) and malonate-Co-A ligase ACSF3 (ACSF3) were downregulated (q < 0.05), whereas the gene expression levels of aquaporin-7 (AQP7), long-chain-fatty-acid-CoA ligase (ACSL), elongation of very long chain fatty acids protein (ELOVL) and fatty acid desaturase 1 (FADS1) were upregulated (q < 0.05), indicating the inhibition of fat catabolism, fat anabolism, fatty acid oxidation, glucose (GLU) intake and SFA synthesis and the promotion of glycerinum (GLY) transportation and PUFA synthesis. Additional findings showed that the gene expression levels of leptin (LEP), adenosine 5'-monophosphate-activated protein kinase (AMPK) and phosphatidylinositol 3-kinase (PI3K) were upregulated (q < 0.05), whereas the gene expression levels of malonyl-CoA decarboxylase (MCD), sterol regulatory element-binding protein 1 (SREBF1), mammalian target of rapamycin (mTOR) and serine/threonine-protein kinase (AKT) were downregulated (q < 0.05), indicating that fat metabolism in the subcutaneous fat of yaks under ES was mainly regulated by AMPK signaling and mTOR and PI3K-AKT signaling were also involved. Energy consumption was inhibited in the subcutaneous fat itself. This study can provide a theoretical basis for the healthy breeding and genetic breeding of yaks.

Grazing exclosures solely are not the best methods for sustaining alpine grasslands.

BACKGROUND: Grazing is widely regarded as a critical factor affecting the vegetation structure, productivity and nutritional value of natural grasslands. To protect and restore degraded grasslands, non-grazed exclosures are considered as a valuable tool. However, it is not clear whether long term non-grazed exclosures of grazers can improve the condition and nutritional value of vegetation and soil properties. METHODS: We have compared the impact of long-term non-grazed and continuous grazed management strategy on vegetation structure, nutritional values and soil properties of alpine meadow of the Qinghai-Tibet Plateau by field investigation (11-13 years) and indoor analysis during 2015-2017. RESULTS: Our results showed that long-term non-grazed exclosures clearly increased the aboveground biomass and coverage of plant functional types. Long-term non-grazed exclosures improved the development of all vegetation types, except NG (GG, grass species type; SG, sedge species type; LG, leguminous species type; FG, forbs species type and NG, noxious species type). Long-term non-grazed exclosures significantly improved all six measured soil properties (TN, total nitrogen; TP, total phosphorus; TK, total potassium; AN, available nitrogen; AP, available phosphorus and AK, available potassium) in 0-10 cm soil layer, considerable effect on the improvement of all measured soil properties, except TK in 10-20 cm soil layer and all measured soil properties, except TN and TK in 20-30 cm soil layer were observed. However, long-term non-grazed exclosures significantly decreased biodiversity indicators i.e., species richness, Shannon diversity index and Evenness index of vegetation. A substantial decrease in the density, biodiversity and nutritional values (CP (crude protein), IVTD (in vitro ture digestibility) and NDF (neutral detergent fiber)) of all vegetation types, except NG were recorded. While a downward trend in aboveground biomass and all measured soil properties except TP and TK were observed during 2015-2017 in alpine meadows due to long-term grazed treatment. The density, diversity and nutritional value (CP and IVTD) of long-term non-grazed alpine meadows showed a downward trend over time (2015-2017). By considering the biodiversity conservation and grassland livestock production, long-term non-grazed exclosures are not beneficial for the improvement of density, biodiversity and nutritional values of plant functional types. Thus, our study suggests that rotational non-grazed and grazed treatment would be a good management strategy to restore and improve the biodiversity and nutritional values of plant functional types in natural grassland ecosystems.

Characterization of the complete mitochondrial genome of the Tianjun yak (Bos grunniens).

Tianjun yak (Bos grunniens) is a yak breed with strong adaptation to the high-elevation, cold and anoxic environments. Its complete mitochondrial genome is 16,323 bp long with an asymmetric base composition, and harbors the 37 typical mitochondrial genes and one noncoding control region. The PCGs are initiated with the typical ATA or ATG codons, and are terminated with TAA, TAG or the incomplete stop codon T--. Phylogenetic analysis suggests that Tianjun yak is most closely related to the polled yak.

Catalytic asymmetric construction of 3,3'-spirooxindoles fused with seven-membered rings by enantioselective tandem reactions.

The first catalytic asymmetric construction of a spirooxindole scaffold incorporated with a seven-membered benzodiazepine moiety has been established by a three-component (isatin, 1,2-phenylenediamine, cyclohexane-1,3-dione) tandem reaction catalyzed by a chiral phosphoric acid. Structurally complex spirobenzodiazepine oxindoles with one quaternary stereogenic center are obtained in high yield with excellent enantioselectivity (up to 99% yield, enantiomeric ratio>99.5:0.5). This approach takes advantage of organocatalytic asymmetric tandem reactions to efficiently construct the structurally rigid spirobenzodiazepine oxindole architecture with high enantiopurity in a single transformation, which involves a cascade enamine-imine formation/intramolecular Mannich reaction sequence.