Transmission of fungi and protozoa under grazing conditions from lactating yaks to sucking yak calves in early life.

Microbiota from mothers is an essential source of microbes in early-life rumen microbiota, but the contribution of microbiota from different maternal sites to the rumen microbiota establishment in neonates needs more data. To fill this gap, we collected samples from the mouth, teat skin, and rumen of lactating yaks and from the rumen of sucking calves concomitantly on seven occasions between days 7 and 180 after birth under grazing conditions. We observed that the eukaryotic communities clustered based on sample sites, except for the protozoal community in the teat skin, with negative correlations between fungal and protozoal diversities in the rumen of calves. Furthermore, fungi in the dam's mouth, which is the greatest source of the calf's rumen fungi, accounted for only 0.1%, and the contribution of the dam's rumen to the calf's rumen fungi decreased with age and even disappeared after day 60. In contrast, the average contribution of the dam's rumen protozoa to the calf's rumen protozoa was 3.7%, and the contributions from the dam's teat skin (from 0.7 to 2.7%) and mouth (from 0.4 to 3.3%) increased with age. Thus, the divergence in dam-to-calf transmissibility between fungi and protozoa indicates that the foundation of these eukaryotic communities is shaped by different rules. This study provides the first measurements of the maternal contribution to the fungal and protozoal establishment in the rumen of sucking and grazing yak calves in early life, which could be beneficial for future microbiota manipulation in neonatal ruminants. KEY POINTS: • Dam to calf transfer of rumen eukaryotes occurs from multiple body sites. • A minor proportion of rumen fungi in calves originated from maternal sites. • The inter-generation transmission between rumen fungi and protozoa differs.

Age-dependent variations in rumen bacterial community of Mongolian cattle from weaning to adulthood.

BACKGROUND: Rumen microbes play an important role in ruminant energy supply and animal performance. Previous studies showed that the rumen microbiome of Mongolian cattle has adapted to degrade the rough forage to provide sufficient energy to tolerate the harsh desert ecological conditions. However, little is known about the succession of rumen microbes in different developmental stages of post-weaning Mongolian cattle. METHODS: Here, we examined the succession of the rumen microbial composition and structure of 15 post-weaning Mongolian cattle at three developmental stages i.e., 5 months (RM05), 18 months (RM18) and, 36 months (RM36) by using the 16S rRNA gene sequencing method. RESULTS: We did not find any age-dependent variations in the ruminal concentrations of any volatile fatty acid (VFA) of Mongolian cattle. The diversity of the rumen bacterial community was significantly lower in RM05 group, which reached to stability with age. Bacteroidetes and Firmicutes were the two dominant phyla among all age groups. Phylum Actinobacteria was significantly higher in RM05 group, phyla Spirochaetes, and Tenericutes were highly abundant in RM18 group, and phyla Proteobacteria and Epsilonbacteraeota were enriched in RM36 group. Genera Prevotella_1, Bacteroides, and Bifidobacterium were abundant in RM05 group. The short chain fatty acid (SCFA) producing bacteria Rikenellaceae_RC9_gut_group showed high abundance in RM18 group and fiber degrading genus Alloprevotella was highly abundant in RM36 group. Random forest analysis identified Alloprevotella, Ileibacterium, and Helicobacter as important age discriminatory genera. In particular, the genera Ruminococcaceae_UCG-005, Bacteroides, Saccharofermentans, and Fibrobacter in RM05, genera [Eubacterium] coprostanoligenes_group, Erysipelotrichaceae_UCG-004, Helicobacter, Saccharofermentans, Papillibacter, and Turicibacter in RM18, and genera Rikenellaceae_RC9_gut_group, Lachnospiraceae_AC2044_group, and Papillibacter in RM36 showed the top interactions values in the intra-group interaction network. CONCLUSIONS: The results showed that rumen microbiota of Mongolian cattle reached to stability and maturity with age after weaning. This study provides some theoretical evidence about the importance of functional specific rumen bacteria in different age groups. Further studies are needed to determine their actual roles and interactions with the host.

Hypothalamic regulation of energy homoeostasis when consuming diets of different energy concentrations: comparison between Tibetan and Small-tailed Han sheep.

Seasonal energy intake of Tibetan sheep on the harsh Qinghai-Tibetan Plateau (QTP) fluctuates greatly and is often well below maintenance requirements. The aim of this study was to gain insight into how the hypothalamus regulates energy homoeostasis in Tibetan sheep. We compared Tibetan and Small-tailed Han sheep (n 24 of each breed), which were each allocated randomly into four groups and offered one of four diets that differed in digestible energy densities: 8·21, 9·33, 10·45 and 11·57 MJ/kg DM. Sheep were weighed every 2 weeks, and it was assumed that the change in body weight (BW) reflected the change in energy balance. The arcuate nucleus of the hypothalamus in Tibetan sheep had greater protein expressions of neuropeptide Y (NPY) and agouti-related peptide (AgRP) when in negative energy balance, but lesser protein expressions of proopiomelanocortin (POMC) and cocaine and amphetamine-regulated transcript (CART) when in positive energy balance than Small-tailed Han sheep. As a result, Tibetan sheep had a lesser BW loss when in negative energy balance and stored more energy and gained more BW when in positive energy balance than Small-tailed Han sheep with the same dietary intake. Moreover, in the hypothalamic adenosine monophosphate-activated protein kinase (AMPK) regulation pathway, Tibetan sheep had greater adenosine monophosphate-activated protein kinase-α 2 protein expression than Small-tailed Han sheep, which supported the premise of a better ability to regulate energy homoeostasis and better growth performance. These differences in the hypothalamic NPY/AgRP, POMC/CART and AMPK pathways between breeds conferred an advantage to the Tibetan over Small-tailed Han sheep to cope with low energy intake on the harsh QTP.

Changes in rumen microbial community composition in yak in response to seasonal variations.

AIMS: Yak is a dominant ruminant, well adapted to grazing on pasture year around in the harsh climate of the 3000-meter-high Qinghai-Tibetan Plateau. The complex microbial community that resides within the yak rumen is responsible for fermentation and contributes to its climatic adaptation. This study aimed to characterize the rumen microbiota responses to wide seasonal variations, especially those necessary for survival in the cold seasons. METHODS AND RESULTS: In the present study, we performed 16s rRNA gene sequencing to investigate the seasonal variations in microbiota composition, diversity and associated volatile fatty acids (VFAs) in yak rumen. The results showed that rumen microbiota were dominated by Bacteroides (72.13%-78.54%) and Firmicutes; the relative abundance of Firmicutes was higher in summer (17.44%) than in winter (10.67%; p < 0.05). The distribution of taxa differed among spring, summer and winter rumen communities (PERMANOVA, p = 0.001), whereas other taxa (e.g., Fibrobacter, Verrucomicrobia, Anaerostipes and Paludibacter), which could potentially help overcome harsh climate conditions were observed in higher abundance during the cold spring and winter seasons. The highest total VFA concentration in the yak rumen was obtained in summer (p < 0.05), followed by spring and winter, and both positive and negative correlations between VFAs and specific genera were revealed. CONCLUSIONS: Microbiota in yak rumen appear to be highly responsive to seasonal variations. Considering environmental factors, we suggest that seasonal adaptation by microbial communities in rumen enables their hosts to survive seasonal scarcity and cold stress in the spring and winter. SIGNIFICANCE AND IMPACT OF STUDY: The present study furthers our understanding of how microbial adaptation to seasonal variations in nutrient availability and climate may function in high plateau ruminants, providing insights into the tripartite relationship between the environment, host and microbiota.

A comparative transcriptome and proteomics study of post-partum ovarian cycle arrest in yaks (Bos grunniens).

Post-partum ovarian cycle arrest is the main factor affecting yak reproductive efficiency. There are few reports regarding the molecular regulatory mechanism of post-partum oestrus at transcriptome and proteome levels in yaks. Our previous studies focussed on the ovaries of yaks with post-partum ovarian cycle arrest and post-partum oestrus yaks. In this study, RNA sequencing transcriptomic study was combined with quantitative proteomic analyses to identify post-partum ovarian cycle-related genes and proteins. Consequently, 1,149 genes and 24 proteins were found to be up- or downregulated during post-partum oestrus. The analysis of differentially regulated genes identified three gene or protein pairs that were synchronously upregulated and no gene or protein pairs that were synchronously downregulated, suggesting that these upregulated genes may regulate the post-partum ovarian cycle. The functional classification of these differentially expressed genes and proteins indicated their connection with the oocyte meiosis, the oestrogen signalling pathway, the progesterone-mediated oocyte maturation and the gonadotrophin-releasing hormone (GnRH) signalling pathway. In this study, a total of six genes and two proteins involved in the oocyte meiosis, the oestrogen signalling pathway, the progesterone-mediated oocyte maturation and the GnRH signalling pathway were identified. The CSNK1A1, M91_09723, M91_11326, M91_21439, M91_19073, SHC2, Atf6b, M91_03062, HSPCA and calmodulin could regulate oestrus, respectively, in the post-partum so as to control the anoestrus status.

Long-term active restoration of extremely degraded alpine grassland accelerated turnover and increased stability of soil carbon.

Soil nutrient contents and organic carbon (C) stability are key indicators for restoration of degraded grassland. However, the effects of long-term active restoration of extremely degraded grassland on soil parameters have been equivocal. The aims of this study were to evaluate the impact of active restoration of degraded alpine grassland on: (a) soil organic matter (SOM) mineralization; and (b) the importance of biotic factors for temperature sensitivity (Q10 ) of SOM mineralization. Soils were sampled from intact, degraded and restored alpine grasslands at altitudes ranging between 3,900 and 4,200 m on the Tibetan Plateau. The samples were incubated at 5, 15 and 25°C, and Q10 values of SOM mineralization were determined. Structural equation modeling was used to evaluate the importance of vegetation, soil physico-chemical properties and microbial parameters for Q10 regulation. The Q10 of N mineralization was similar among intact, degraded and restored soils (0.84-1.24) and was higher in topsoil (1.09) than in subsoil (0.92). The best predictive factor of CO2 -Q10 for intact grassland was microbial biomass, for degraded grassland was basal microbial respiration, and for restored grassland was soil bulk density. Restoration by planting vegetation decreased the Q10 of SOM mineralization as soil bulk density, the most important negative predictor, increased in restored grassland. The Q10 of SOM mineralization in topsoil was 14% higher than in subsoil because of higher microbial abundance and exo-enzyme activities. The NH4 + content was greatest in intact soil, while NO3 - content was greatest in degraded soil. The SOM mineralization rate decreased with grassland degradation and increased after long-term (>10 years) restoration. In conclusion, extremely degraded grassland needs proper long-term management in active restoration projects, especially for improvement of soil nutrients in a harsh environment.

Comparison between Tibetan and Small-tailed Han sheep in adipocyte phenotype, lipid metabolism and energy homoeostasis regulation of adipose tissues when consuming diets of different energy levels.

This study aimed to gain insight into how adipose tissue of Tibetan sheep regulates energy homoeostasis to cope with low energy intake under the harsh environment of the Qinghai-Tibetan Plateau (QTP). We compared Tibetan and Small-tailed Han sheep (n 24 of each breed), all wethers and 1·5 years of age, which were each divided randomly into four groups and offered diets of different digestible energy (DE) densities: 8·21, 9·33, 10·45 and 11·57 MJ DE/kg DM. When the sheep lost body mass and were assumed to be in negative energy balance: (1) adipocyte diameter in subcutaneous adipose tissue was smaller and decreased to a greater extent in Tibetan than in Small-tailed Han sheep, but the opposite occurred in the visceral adipose tissue; (2) Tibetan sheep showed higher insulin receptor mRNA expression and lower concentrations of catabolic hormones than Small-tailed Han sheep and (3) Tibetan sheep had lower capacity for glucose and fatty acid uptake than Small-tailed Han sheep. Moreover, Tibetan sheep had lower AMPKα mRNA expression but higher mammalian target of rapamycin mRNA expression in the adipocytes than Small-tailed Han sheep. We concluded that Tibetan sheep had lower catabolism but higher anabolism in adipose tissue and reduced the capacity for glucose and fatty acid uptake to a greater extent than Small-tailed Han sheep to maintain energy homoeostasis when in negative energy balance. These responses provide Tibetan sheep with a high ability to cope with low energy intake and with the harsh environment of the QTP.

Tibetan sheep have a high capacity to absorb and to regulate metabolism of SCFA in the rumen epithelium to adapt to low energy intake.

The nutritional intake of Tibetan sheep on the harsh Qinghai-Tibetan Plateau is often under maintenance requirements, especially during the long, cold winter. However, they have adapted well and even thrive under these conditions. The aim of the present study was to gain insight into how the rumen epithelium of Tibetan sheep has adapted to the consumption of low-energy-level diets. For this purpose, we compared Tibetan and small-tailed Han sheep (n 24 of each breed, all wethers and 1·5 years of age), which were divided randomly into one of four groups and offered ad libitum diets of different digestible energy (DE) densities: 8·21, 9·33, 10·45 and 11·57 MJ DE/kg DM. The Tibetan sheep had higher rumen concentrations of total SCFA, acetate, butyrate and iso-acids but lower concentrations of propionate than small-tailed Han sheep. The Tibetan sheep had higher absorption capability of SCFA due to the greater absorption surface area and higher mRNA expression of the SCFA absorption relative genes than small-tailed Han sheep. For the metabolism of SCFA in the rumen epithelium, the small-tailed Han sheep showed higher utilisation of the ketogenesis pathway than Tibetan sheep; however, Tibetan sheep had greater regulation capacity in SCFA metabolism pathways. These differences between breeds allowed the Tibetan sheep to have greater capability of absorbing SCFA and better capacity to regulate the metabolism of SCFA, which would allow them to cope with low energy intake better than small-tailed Han sheep.

Rumen parameters of yaks (Bos grunniens) and indigenous cattle (Bos taurus) grazing on the Qinghai-Tibetan Plateau.

Yaks and indigenous Qaidam cattle and cattle-yak crosses (C × Y) graze on the Qinghai-Tibetan Plateau (QTP) throughout the year, but yaks are raised at higher elevations than cattle. Yaks do not receive supplementary feed whereas cattle require supplementary feed during harsh winter. We hypothesized that yaks would cope with the severe conditions of the QTP better than cattle and utilize the pasture more efficiently. We also hypothesized that differences between species would be pronounced in winter, when conditions are particularly harsh. To test these hypotheses, seasonal rumen fluid parameters of yaks, C × Y and cattle (n = 3 for each) were examined. Rumen fluid was collected in summer and winter from each genotype 2, 5 and 12 hr after a day of grazing. Concentrations of total volatile fatty acid (VFA), acetate, propionate, isobutyrate and isovalerate were greater in yaks than in cattle in summer (p < 0.05), while propionate concentration was lower in yaks than in cattle in winter (p < 0.05). Concentrations of ammonia and urea were greater (p < 0.001) in yaks than in cattle (p < 0.001) in summer, whereas, concentrations of free amino acids (AA) were greater in cattle than in yaks in summer and winter (p < 0.001). Concentrations of total VFA, acetate, propionate and butyrate decreased linearly (p < 0.05), whereas concentrations of isobutyrate and isovalerate increased linearly for yak and C × Y with sampling time (p < 0.05) in summer. In summer, concentrations of isobutyrate and isovalerate were greater in yaks than in cattle (p < 0.05). In conclusion, rumen fermentation characteristics of yaks showed that they coped better than cattle or C × Y in the harsh climate as we hypothesized. However, in contrast to our hypothesis, this emerged only in summer, when pasture was plentiful and not in winter, when pasture was scarce.

Effect of dietary energy on digestibilities, rumen fermentation, urinary purine derivatives and serum metabolites in Tibetan and small-tailed Han sheep.

Tibetan sheep are indigenous to the Qinghai-Tibetan Plateau, graze the grassland all year round without supplementation and are well-adapted to the harsh conditions. Small-tailed Han sheep were introduced to the plateau and are raised mainly in feedlots. Based on their different backgrounds, we hypothesized that the ability to cope with poor diets would be better in Tibetan than in Han sheep. To test our prediction, we examined the effect of dietary energy on apparent digestibilities, rumen fermentation, urinary purine derivatives and serum metabolites by using a 4 × 4 Latin square design in each sheep breed. Four diets were formulated to be low in crude protein (~7%) but to differ in metabolizable energy concentration. Average daily gain was greater in Tibetan than in Han sheep (p < 0.01) and increased linearly with an increase in energy intake (p < 0.001). The digestibilities of dry matter, organic matter, gross energy, and neutral and acid detergent fibres were greater in Tibetan than in Han sheep (p < 0.05). Ruminal pH was lower (p < 0.05), while volatile fatty acids (VFAs), urea-N, ammonia-N and soluble protein-N concentrations were higher (p < 0.05) in Tibetan than in Han sheep. As a molar proportion of total VFA, acetate decreased (p < 0.001) with an increase in dietary energy whereas propionate and butyrate increased (p < 0.05). Urinary purine derivative excretion was greater in Tibetan than in Han sheep (p < 0.01), as was microbial nitrogen production; both parameters increased with dietary energy (p < 0.01). Serum concentrations of glucose, insulin and insulin-like growth factor-1 increased (p < 0.05) as energy level increased, while non-esterified fatty acids and growth hormone decreased (p < 0.05). It was concluded that Tibetan sheep were better able to cope with low-protein, low-energy diets and, consequently, our prediction was supported.

Bacterial communities related to 3-nitro-1-propionic acid degradation in the rumen of grazing ruminants in the Qinghai-Tibetan Plateau.

The objectives of this current study were to characterize the overall rumen bacterial community in grazing yak and two sheep species (Tibetan and Small Tail Han sheep) reared in the unique environmental conditions of the Qinghai-Tibetan Plateau, as well as the bacterial community associated with the detoxification of a phytotoxin, 3-nitro-1-propionic acid (NPA), during in vitro culture with 4.2 mM NPA. Using 16S rRNA gene high-throughput sequencing, it was found that the yak rumen harbored populations showing a higher bacterial diversity compared to Tibetan sheep. The rumen bacterial community in the three ruminant species differed from each other. PICRUSt analysis identified that the pathway involved in nitrogen metabolism was enriched in Tibetan sheep while that related to fatty acid biosynthesis was over-represented in the yak. The methane metabolism pathway was dominant in bacterial populations from the Small Tail Han sheep. Comparisons between freshly collected rumen fluid and populations subjected to consecutive 72 h batch cultures revealed substantial decreases in alpha diversity in populations cultured with NPA. Moreover, the relative abundances of some bacterial taxa changed significantly, with increased abundance of Proteobacteria and Actinobacteria. In addition, the overall community structure of the bacterial population in the freshly collected ruminal fluid was clearly different than that within populations observed in the 72 h batch cultures likely due to the impact of NPA treatments and the more restrictive growth conditions of the culture medium. In regard to PICRUSt analysis, the methane metabolism pathway became scarce in Tibetan and Small Tail Han sheep, whereas the energy and carbohydrate metabolic pathways such as nitrogen metabolism, ABC transporters and glycolysis/gluconeogenesis were found to be maintained across all populations. Results from the present study provide new information on the bacterial and functional composition within ruminal populations adapted to three economically important grazing ruminant species prominent on the Qinghai-Tibetan Plateau. The results further reveal that effects of NPA treatment on community structure can have an impact not only the metabolism of NPA but on other digestive functions as well.

Methanogen Diversity in Indigenous and Introduced Ruminant Species on the Tibetan Plateau.

Host factors are regarded as important in shaping the archaeal community in the rumen but few controlled studies have been performed to demonstrate this across host species under the same environmental conditions. A study was designed to investigate the structure of the methanogen community in the rumen of two indigenous (yak and Tibetan sheep) and two introduced domestic ruminant (cattle and crossbred sheep) species raised and fed under similar conditions on the high altitude Tibetan Plateau. The methylotrophic Methanomassiliicoccaceae was the predominant archaeal group in all animals even though Methanobrevibacter are usually present in greater abundance in ruminants globally. Furthermore, within the Methanomassiliicoccaceae family members from Mmc. group 10 and Mmc. group 4 were dominant in Tibetan Plateau ruminants compared to Mmc. group 12 found to be highest in other ruminants studied. Small ruminants presented the highest number of sequences that belonged to Methanomassiliicoccaceae compared to the larger ruminants. Although the methanogen community structure was different among the ruminant species, there were striking similarities between the animals in this environment. This indicates that factors such as the extreme environmental conditions and diet on the Tibetan Plateau might have a greater impact on rumen methanogen community compared to host differences.

Fiber degradation potential of natural co-cultures of Neocallimastix frontalis and Methanobrevibacter ruminantium isolated from yaks (Bos grunniens) grazing on the Qinghai Tibetan Plateau.

Several natural anaerobic fungus-methanogen co-cultures have been isolated from rumen and feces source of herbivores with strong fiber degrading ability. In this study, we isolated 7 Neocallimastix with methanogen co-cultures from the rumen of yaks grazing on the Qinghai Tibetan Plateau. Based on morphological characteristics and internal transcribed spacer 1 sequences (ITS1), all the fungi were identified as Neocallimastix frontalis. The co-cultures were confirmed as the one fungus - one methanogen pattern by the PCR-denatured gradient gel electrophoresis (DGGE) assay. All the methanogens were identified as Methanobrevibacter ruminantium by 16s rRNA gene sequencing. We investigated the biodegrading capacity of the co-culture (N. frontalis + M. ruminantium) Yaktz1 on wheat straw, corn stalk and rice straw in a 7 days-incubation. The in vitro dry matter digestibility (IVDMD), acid detergent fiber digestibility (ADFD) and neural detergent fiber digestibility (NDFD) values of the substrates in the co-culture were significantly higher than those in the mono-culture N. frontalis Yaktz1. The co-culture exhibited high polysaccharide hydrolase (xylanase and FPase) and esterase activities. The xylanase in the co-culture reached the highest activity of 12500 mU/ml on wheat straw at the day 3 of the incubation. At the end of the incubation, 3.00 mmol-3.29 mmol/g dry matter of methane were produced by the co-culture. The co-culture also produced high level of acetate (40.00 mM-45.98 mM) as the end-product during the biodegradation. Interestingly, the N. frontalis Yaktz1 mono-culture produced large amount of lactate (8.27 mM-11.60 mM) and ethanol (163.11 mM-242.14 mM), many times more than those recorded in the previously reported anaerobic fungi. Our data suggests that the (N. frontalis + M. ruminantium) Yaktz1 co-culture and the N. frontalis Yaktz1 mono-culture both have great potentials for different industrial use.

Importance of functional ingredients in yak milk-derived food on health of Tibetan nomads living under high-altitude stress: a review.

Tibetan nomads have lived since ancient times in the unique and harsh environment of the Qinghai-Tibetan Plateau with average altitudes over 4000 m. These people have been able to live and multiply healthily over numerous generations under the extreme stress of high-altitude environment, including cold, hypoxia, and strong ultraviolet radiation, and with a simple diet devoid of vegetables and fruits for most of the year. Their survival depends heavily on yak milk, and its products comprise the main portion of their daily diet. In this review, yak milk and its derived products are examined in detail and compared with milk from other ruminant species. Yak milk products seem to be particularly rich in functional and bioactive components, which may play a role in maintaining the health status of Tibetan nomads. This includes particular profiles of amino acids and fatty acids, and high levels of antioxidant vitamins, specific enzymes, and bacteria with probiotic activity (yoghurt is the main food). Based on that, it is proposed that the Tibetan nomads have developed a nutritional mechanism adapted to cope with the specific challenges posed by the environment of the world's highest plateau. Systematic studies are required to demonstrate this in a more mechanistic way.

Characterization of bovine ruminal and equine cecal microbial populations enriched for enhanced nitro-toxin metabolizing activity.

The nitrotoxins 3-nitro-1-propionic acid (NPA) and 3-nitro-1-propanol (NPOH) are produced by a wide variety of leguminous plants, including over 150 different species and varieties of Astragalus potentially grazed by livestock. These toxins are known to be detoxified by at least one ruminal bacterium but detoxification by bacteria from other gut habitats is not known. In the present study, mixed populations of bovine ruminal and equine cecal microbes were enriched for NPA-metabolizing bacteria via consecutive 24-72 h culture in a basal minimal rumen fluid-based medium supplemented with 4.2 mM NPA and H2 as the energy source. Rates of NPA metabolism by the respective populations increased from 58.4 ± 4.8 and 8.6 ± 11.6 nmol NPA/mL per h during initial culture to 88.9 ± 30.6 and 50.2 ± 30.9 nmol NPA/mL per h following enrichment. Results from 3-tube most probable number tests indicated that numbers of NPA-degrading microbes increased 2.1 and 1.8 log10 units during enrichment from numbers measured pre-enrichment (3.9 × 10³ and 4.3 × 10¹ cells/mL for ruminal and equine cecal populations, respectively). Hydrogen, formate, and to a lesser extent, DL-lactic acid, served as electron donors to the enriched populations and CO2 or formate were needed to maintain high rates of NPA-metabolism. The NPA-enriched populations were able to metabolize nitrate which, being a preferred electron acceptor, was antagonistic to NPA metabolism. Supplemental NPA was inhibitory to methanogenesis. Fermentation balance estimates indicated that only 47.6% of carbon available in potential substrates was recovered in headspace CO2, volatile fatty acids or unmetabolized NPA after 72 h incubation of NPA-enriched populations that had metabolized 98% of 8.4 mM added NPA. Overall, these results reveal low level carriage of NPA-metabolizing, CO2 or formate-requiring bacterial populations in the equine cecum yet support the concept that Denitrobacterium detoxificans-like organisms may well be the functional agents of NPA and NPOH detoxification in the populations studied here.

Linkages between rumen microbiome, host, and environment in yaks, and their implications for understanding animal production and management.

Livestock on the Qinghai-Tibetan Plateau is of great importance for the livelihood of the local inhabitants and the ecosystem of the plateau. The natural, harsh environment has shaped the adaptations of local livestock while providing them with requisite eco-services. Over time, unique genes and metabolic mechanisms (nitrogen and energy) have evolved which enabled the yaks to adapt morphologically and physiologically to the Qinghai-Tibetan Plateau. The rumen microbiota has also co-evolved with the host and contributed to the host's adaptation to the environment. Understanding the complex linkages between the rumen microbiota, the host, and the environment is essential to optimizing the rumen function to meet the growing demands for animal products while minimizing the environmental impact of ruminant production. However, little is known about the mechanisms of host-rumen microbiome-environment linkages and how they ultimately benefit the animal in adapting to the environment. In this review, we pieced together the yak's adaptation to the Qinghai-Tibetan Plateau ecosystem by summarizing the natural selection and nutritional features of yaks and integrating the key aspects of its rumen microbiome with the host metabolic efficiency and homeostasis. We found that this homeostasis results in higher feed digestibility, higher rumen microbial protein production, higher short-chain fatty acid (SCFA) concentrations, and lower methane emissions in yaks when compared with other low-altitude ruminants. The rumen microbiome forms a multi-synergistic relationship among the rumen microbiota services, their communities, genes, and enzymes. The rumen microbial proteins and SCFAs act as precursors that directly impact the milk composition or adipose accumulation, improving the milk or meat quality, resulting in a higher protein and fat content in yak milk and a higher percentage of protein and abundant fatty acids in yak meat when compared to dairy cow or cattle. The hierarchical interactions between the climate, forage, rumen microorganisms, and host genes have reshaped the animal's survival and performance. In this review, an integrating and interactive understanding of the host-rumen microbiome environment was established. The understanding of these concepts is valuable for agriculture and our environment. It also contributes to a better understanding of microbial ecology and evolution in anaerobic ecosystems and the host-environment linkages to improve animal production.

Seed rain and its relationship with above-ground vegetation of degraded Kobresia meadows.

Seed rain is a crucial element in vegetation regeneration, but has been rarely studied in high altitude regions, particularly degraded Kobresia meadow. Weed infestation is a distinctive feature of pasture degradation in Kobresia meadows on the Tibetan plateau, the ecological mechanism of which is closely related with vegetation's seed rain. In this paper we assess the effect of vegetation degradation on seed rain and consider its implication for restoration of degraded Kobresia meadows in the headwater area of Yellow river, through analysis of seed species composition, number of seeds landing per m(2) of soil surface, and their relationship with above ground vegetation. Vegetation degradation had an impact on the species composition and numbers of seeds in seed rain and their relationship with above-ground vegetation. Within the un-degraded meadow, which provided a closed vegetation cover, 35 % of the seed rain was of sedge and gramineae species. However, within the degraded meadows, as the extent of degradation increased, so the total number of seeds m(-2) increased, with those derived from sedge and gramineae species forming a declining proportion of the total. Degradation of Kobresia meadow on the Tibetan plateau is exacerbated by the seed input of weed species (such as Oxytropis ochrocephala, Carum carvi, Aconitum pendulum, Pedicularis kansuensis in this study). Therefore, a major priority for the restoration of such degraded meadows should be the elimination of these weeds from the above ground vegetation by human intervention.

Seasonal variations in the fatty acid profile of milk from yaks grazing on the Qinghai-Tibetan plateau.

An experiment was conducted to study the seasonal changes in the fatty acid profile of milk from yaks (Bos grunniens) when kept at altitudes of 3000 m above sea level (a.s.l.) and higher. Data and samples were collected in summer (July), autumn (September), winter (November) and spring (March) from ten lactating yaks (four in spring). The yaks grazed pastures adjacent to the farm building throughout the year. In spring only they received 0·6 kg crop by-products per day (dry matter basis). Fresh alpine grasses, available in summer and autumn, showed high concentrations of α-linolenic acid (46-51 g/100 g lipids) compared with the dry, yellow vegetation of winter and spring (16 g/100 g lipids). In autumn and summer, the milk fat had higher concentrations of polyunsaturated fatty acids than in winter. These polyunsaturated fatty acids were comprised of vaccenic acid, rumenic acid and α-linolenic acid, which are all considered beneficial to human health. The rare fatty acid, γ-linolenic acid, was also detected in yak milk, especially in the milk obtained in spring. The results suggest that yak milk, which is the most important basic food of the Tibetan herders, has the most favourable fatty acid profile when yaks grazed green pasture, which also corresponds to the period of highest milk production.

Unexpected no significant soil carbon losses in the Tibetan grasslands due to rodent bioturbation.

The Tibetan grasslands store 2.5% of the Earth's soil organic carbon. Unsound management practices and climate change have resulted in widespread grassland degradation, providing open habitats for rodent activities. Rodent bioturbation loosens topsoil, reduces productivity, changes soil nutrient conditions, and consequently influences the soil organic carbon stocks of the Tibetan grasslands. However, these effects have not been quantified. Here, using meta-analysis and upscaling approaches, we found that rodent bioturbation impacts on the Tibetan grassland soil organic carbon contents were depth-dependent, with significant (P < 0.001) decreasing of 24.4% in the topsoil (0 to 10 cm) but significant (P < 0.05) increasing of 35.9% in the deeper soil layer (40 to 50 cm), and nonsignificant changes in other soil layers. The depth-dependent responses in soil organic carbon content were closely associated with rodent tunnel burrowing, foraging, excrement deposition, and mixing of the upper and deeper soil layers. Rodent bioturbation had shown nonsignificant impacts on soil bulk density, independent of soil layer. Tibetan grasslands totally lose -35.2 Tg C yr-1 (95% CI: -48.5 to -21.1 Tg C yr-1) and -32.9 Tg C yr-1 (-54.2 to -8.6 Tg C yr-1) due to rodent bioturbation in the 0 to 10 or 0 to 30 cm soil layer, while no significant net loss was found over the 0 to 90 cm layer. Our findings highlight the importance of considering depth-dependent factors to robustly quantify the net changes in the terrestrial soil organic carbon stocks resulting from disturbances such as rodent bioturbation.

A comparison of average daily gain, apparent digestibilities, energy balance, rumen fermentation parameters, and serum metabolites between yaks (Bos grunniens) and Qaidam cattle (Bos taurus) consuming diets differing in energy level.

Yaks (Bos grunniens), indigenous to the harsh Qinghai-Tibetan Plateau, are well adapted to the severe conditions, and graze natural pasture without supplements all year round. Qaidam cattle (Bos taurus), introduced to the Qinghai-Tibetan Plateau 1,700 years ago, are raised at a lower altitude than yaks, provided with shelter at night and offered supplements in winter. Based on their different backgrounds, we hypothesized that yaks have lower energy requirements for maintenance than cattle. To test this hypothesis, we measured average daily gain (ADG), apparent digestibilities, energy balance, rumen fermentation parameters, and serum metabolites in growing yaks and cattle offered diets differing in metabolizable energy (ME) levels (6.62, 8.02, 9.42 and 10.80 MJ/kg), but with the same crude protein concentration. Six castrated yaks (155 ± 5.8 kg) and 6 castrated Qaidam cattle (154 ± 8.0 kg), all 2.5 years old, were used in 2 concurrent 4 × 4 Latin square designs. Neutral and acid detergent fiber digestibilities were greater (P < 0.05) in yaks than in cattle, and decreased linearly (P < 0.05) with increasing dietary energy level; whereas, digestibilities of dry matter, organic matter, crude protein and ether extract increased (P < 0.05) linearly with increasing energy level. The ADG was greater (P < 0.001) in yaks than in cattle, and increased (P < 0.05) linearly with increasing energy levels. From the regressions of ADG on ME intake, the estimated ME requirement for maintenance was lower (P < 0.05) in yaks than in cattle (0.43 vs. 0.57 MJ/kg BW0.75). The ratios of digestible energy (DE):gross energy and ME:DE were higher (P < 0.05) in yaks than in cattle, and increased (P < 0.05) linearly with increasing dietary energy level. Ruminal pH decreased (P < 0.05), whereas concentrations of total volatile fatty acids (VFAs) and ammonia increased (P < 0.01) with increasing dietary energy level, and all were greater (P < 0.05) in yaks than in cattle. Concentrations of ruminal acetate and iso-VFAs were greater (P < 0.05), whereas propionate was lower (P < 0.05) in yaks than in cattle; acetate decreased (P < 0.001), whereas butyrate and propionate increased (P < 0.001) linearly with increasing dietary energy level. Serum concentrations of ß-hydroxybutyrate were lower (interaction, P < 0.001) in yaks than in cattle fed diets of 9.42 and 10.80 MJ/kg, whereas non-esterified fatty acids were greater (interaction, P < 0.01) in yaks than in cattle fed diets of 6.62 and 8.02 MJ/kg. Concentrations of serum leptin and growth hormone were greater in yaks than in cattle and serum insulin and growth hormone increased (P < 0.01) linearly with increasing dietary energy level. Our hypothesis that yaks have lower energy requirements for maintenance than cattle was supported. This lower requirement confers an advantage to yaks over Qaidam cattle in consuming low energy diets during the long winter on the Qinghai-Tibetan Plateau.