Dynamic changes of rumen microbiota and serum metabolome revealed increases in meat quality and growth performances of sheep fed bio‑fermented rice straw

Background Providing high-quality roughage is crucial for improvement of ruminant production because it is an essential component of their feed.Our previous study showed that feeding bio-fermented rice straw(BF)improved the feed intake and weight gain of sheep.However,it remains unclear why feeding BF to sheep increased their feed intake and weight gain.Therefore,the purposes of this research were to investigate how the rumen micro-biota and serum metabolome are dynamically changing after feeding BF,as well as how their changes influence the feed intake,digestibility,nutrient transport,meat quality and growth performances of sheep.Twelve growing Hu sheep were allocated into 3 groups:alfalfa hay fed group(AH:positive control),rice straw fed group(RS:negative control)and BF fed group(BF:treatment).Samples of rumen content,blood,rumen epithelium,muscle,feed offered and refusals were collected for the subsequent analysis.Results Feeding BF changed the microbial community and rumen fermentation,particularly increasing(P<0.05)relative abundance of Prevotella and propionate production,and decreasing(P<0.05)enteric methane yield.The histomorphology(height,width,area and thickness)of rumen papillae and gene expression for carbohydrate trans-port(MCT1),tight junction(claudin-1,claudin-4),and cell proliferation(CDK4,Cyclin A2,Cyclin E1)were improved(P<0.05)in sheep fed BF.Additionally,serum metabolome was also dynamically changed,which led to up-regulating(P<0.05)the primary bile acid biosynthesis and biosynthesis of unsaturated fatty acid in sheep fed BF.As a result,the higher(P<0.05)feed intake,digestibility,growth rate,feed efficiency,meat quality and mono-unsaturated fatty acid concentration in muscle,and the lower(P<0.05)feed cost per kg of live weight were achieved by feeding BF.Conclusions Feeding BF improved the growth performances and meat quality of sheep and reduced their feed cost.Therefore,bio-fermentation of rice straw could be an innovative way for improving ruminant production with mini-mizing production costs.

The community structure of Methanomassiliicoccales in the rumen of Chinese goats and its response to a highgrain diet

Background: The newly proposed methanogenic order ‘Methanomassiliicoccales' is the second largest archaeal population in the rumen, second only to the Methanobrevibacter population. However, information is limited regarding the community of this new order in the rumen.Methods: This study used real-time PCR and 454 pyrosequencing to explore the abundance and community composition of Methanomassiliicoccales in the rumen of Chinese goats fed a hay(0% grain, n = 5) or a high grain(65% grain, n = 5) diet.Results: Real-time PCR analysis showed that the relative abundance of Methanomassili coccales(% of total archaea) in the goat rumen was significantly lower in the high-grain-diet group(0.5% ± 0.2%) than that in the hay-diet group(8.2% ± 1.1%, P < 0.05). The pyrosequencing results showed that a total of 208 operational taxonomic units(OTUs) were formed from ten samples at 99% sequence identity. All the sequences were identified as Methanomassiliicoccaceae at the family level, and most of the sequences(96.82% ± 1.64%) were further classified as Group 8, 9, and 10 at the Methanomassiliicoccales genus level in each sample based on the RIM-DB database. No significant differences were observed in the number of OTUs or Chao1's, Shannon's or Pielou's evenness indexes between the hay-and high-grain-diet groups(P ≥ 0.05). PCoA analysis showed that diet altered the community of Methanomassiliicoccales.At the genus level, the relative abundances of Group 10(67.25 ± 12.76 vs. 38.13 ± 15.66, P = 0.012) and Group 4(2.07 ± 1.30 vs. 0.27 ± 0.30, P = 0.035) were significantly higher in the high-grain-diet group, while the relative abundance of Group9 was significantly higher in the hay-diet group(18.82 ± 6.20 vs. 47.14 ± 17.72, P = 0.020). At the species level, the relative abundance of Group 10 sp.(67.25 ± 12.76 vs. 38.13 ± 15.66, P = 0.012) and Group 4 sp. MpT 1(2.07 ± 1.30 vs. 0.27 ± 0.30,P = 0.035) were significantly higher in the high-grain-diet group, while the relative abundance of Group 9 sp. ISO4-G1 was significantly higher in the hay-diet group(12.83 ± 3.87 vs. 42.44 ± 18.47, P = 0.022).Conclusions: Only a few highly abundant phylogenetic groups dominated within the Methanomassiliicoccales community in the rumens of Chinese goats, and these were easily depressed by high-grain-diet feeding. The relatively low abundance suggests a small contribution on the part of Methanomassiliicoccales to the rumen methanogenesis of Chinese goats.

A pH-sensitive supramolecular nanosystem with chlorin e6 and triptolide co-delivery for chemo-photodynamic combination therapy

The combination of Ce6,an acknowledged photosensitizer,and TPL,a natural anticancer agent,has been demonstrated as a useful strategy to reinforce the tumor growth suppression,as well as decrease the systemic side effects compared with their monotherapy.However,in view of the optimal chemo-photodynamic combination efficiency,there is still short of the feasible nanovehicle to steadily co-deliver Ce6 and TPL,and stimuli-responsively burst release drugs in tumor site.Herein,we described the synergistic antitumor performance of a pH-sensitive supramolecular nanosystem,mediated by the host–guest complexing betweenβ-CD and acid pH-responsive amphiphilic co-polymer mPEG-PBAE-mPEG,showing the shell–core structural micelles with the tightβ-CD layer coating.Both Ce6 and TPLwere facilely co-loaded into the spherical supramolecular NPs(TPL+Ce6/NPs)by one-step nanoprecipitation method,with an ideal particle size(156.0 nm),acid pH-responsive drug release profile,and enhanced cellular internalization capacity.In view of the combination benefit of photodynamic therapy and chemotherapy,as well as co-encapsulation in the fabricated pH-sensitive supramolecular NPs,TPL+Ce6/NPs exhibited significant efficacy to suppress cellular proliferation,boost ROS level,lower MMP,and promote cellular apoptosis in vitro.Particularly,fluorescence imaging revealed that TPL+Ce6/NPs preferentially accumulated in the tumor tissue area,with higher intensity than that of free Ce6.As expected,upon 650-nm laser irradiation,TPL+Ce6/NPs exhibited a cascade of amplified synergistic chemo-photodynamic therapeutic benefits to suppress tumor progression in both hepatoma H22 tumor-bearingmice and B16 tumor-bearingmice.More importantly,lower systemic toxicitywas found in the tumor-bearingmice treated with TPL+Ce6/NPs.Overall,the designed supramolecular TPL+Ce6/NPs provided a promising alternative approach for chemo-photodynamic therapy in tumor treatment.

The effect of increased atmospheric temperature and CO_2 concentration during crop growth on the chemical composition and in vitro rumen fermentation characteristics of wheat straw

This experiment was conducted to investigate the effects of increased atmospheric temperature and CO2 concentration during crop growth on the chemical composition and in vitro rumen fermentation characteristics of wheat straw. The field experiment was carried out from November 2012 to June 2013 at Changshu （31°32′93″N, 120°41′88″E） agro-ecological experimental station. A total of three treatments were set. The concentration of CO2 was increased to 500 pmol/mol in the first treatment （CO2 group）. The temperature was increased by 2℃ in the second treatment （TEM group） and the concentration of CO2 and temperature were both increased in the third treatment （CO2 ＋ TEM group）. The mean temperature and concentration of CO2 in control group were 10.5 ℃ and 413μmol/mol. At harvesting, the wheat straws were collected and analyzed for chemical composition and in vitro digestibility. Results showed that dry matter was significantly increased in all three treatments. Ether extracts and neutral detergent fiber were significantly increased in TEM and CO2 ＋ TEM groups. Crude protein was significantly decreased in CO2＋TEM group. In vitro digestibility analysis of wheat straw revealed that gas production was significantly decreased in CO2 and CO2 ＋ TEM groups. Methane production was significantly decreased in TEM and CO2 ＋ TEM groups. Ammonia nitrogen and microbial crude protein were significantly decreased in all three treatments. Total volatile fatty acids were significantly decreased in CO2 and CO2 ＋ TEM groups. In conclusion, the chemical composition of the wheat straw was affected by temperature and CO2 and the in vitro digestibility of wheat straw was reduced, especially in the combined treatment of temperature and CO2.

Production,purific tion,characterization and application of two novel endoglucanases from buffalo rumen metagenome

Background Lignocellulose biomass is the most abundant and renewable material in nature.The objectives of this study were to characterize two endoglucanases Trep Cel3 and Trep Cel4,and determine the effect of the combination of them(1.2 mg Trep Cel3,0.8 mg Trep Cel4)on in vitro rumen fermentation characteristics.In this study,three nature lignocellulosic substrates(rice straw,RS;wheat straw,WS;leymus chinensis,LC)were evaluated for their in vitro digestibility,gas,NH3-N and volatile fatty acid(VFA)production,and microbial protein(MCP)synthesis by adding enzymatic combination.Methods Two endoglucanases’genes were successfully expressed in Escherichia coli(E.coli)BL21(DE3),and enzymatic characteristics were further characterized.The combination of Trep Cel3 and Trep Cel4 was incubated with lignocellulosic substrates to evaluate its hydrolysis ability.Results The maximum enzymatic activity of Trep Cel3 was determined at p H 5.0 and 40℃,while Trep Cel4 was at p H 6.0 and 50℃.They were stable over the temperature range of 30 to 60℃,and active within the p H range of 4.0 to 9.0.The Trep Cel3 and Trep Cel4 had the highest activity in lichenan 436.9±8.30 and 377.6±6.80 U/mg,respectively.The combination of Trep Cel3 and Trep Cel4 exhibited the highest efficiency at the ratio of 60:40.Compared to maximum hydrolysis of Trep Cel3 or Trep Cel4 separately,this combination was shown to have a superior ability to maximize the saccharification yield from lignocellulosic substrates up to 188.4%for RS,236.7%for wheat straw WS,222.4%for LC and 131.1%for sugar beet pulp(SBP).Supplemental this combination enhanced the dry matter digestion(DMD),gas,NH3-N and VFA production,and MCP synthesis during in vitro rumen fermentation.Conclusions The Trep Cel3 and Trep Cel4 exhibited the synergistic relationship(60:40)and significantly increased the saccharification yield of lignocellulosic substrates.The combination of them stimulated in vitro rumen fermentation of lignocellulosic substrates.This combination has the potential to be a feed additive to improve agricultural residues utilization in ruminants.If possible,in the future,experiments in vivo should be carried out to fully evaluate its effect.

瘤胃甲烷菌第七目的研究进展

随着测序技术和体外培养技术的进步,越来越多的未知甲烷菌被发现。近年来,第七个甲烷菌目"Methanomassiliicoccales"(简称Mmc)被发现并建立。这一目甲烷菌在进化关系上与热原体古菌相近,但又存在较远距离,独立成簇。Mmc分布广泛,遍布哺乳动物和昆虫的消化道以及稻田、湿地等环境,但不同来源菌株表现出生境偏好性。Mmc缺少将CO2还原为甲基辅酶M的完整代谢途径,导致它们严格利用氢气还原甲基底物生成甲烷。全面深入地了解Mmc在反刍动物瘤胃中发挥的功能,将有助于新型高效反刍动物甲烷减排策略的提出。因此,本文主要综述了Mmc菌株的分离培养、生理生化和基因组特性及其在瘤胃甲烷生成中的作用。

日粮中添加不同比例微贮棉秆对湖羊瘤胃微生物区系的影响

【目的】本试验旨在利用16SrDNA高通量测序技术研究不同比例微贮棉秆的添加对断奶湖羊瘤胃微生物区系的影响。【方法】选择日龄相近、体重相似的湖羊30只,根据日粮中微贮棉秆的含量随机分为3组:对照组(S0)、50%微贮棉秆组(S50)和100%微贮棉秆组(S100),每组随机屠宰6只分析生长性能,并取瘤胃液进行瘤胃发酵参数和微生物区系分析。【结果】饲喂50%微贮棉秆能够显著提高湖羊日增重和屠宰率(P<0.05)。Bacteroidetes和Firmicutes是湖羊瘤胃的优势菌门,Prevotella和Unclassified Bacteroidales是湖羊瘤胃的优势菌属。日粮中添加100%微贮棉秆可以显著降低湖羊瘤胃菌群的多样性(P<0.05);显著降低Unclassified Bacteroidales和BF311的相对丰度(P<0.05)。三条代谢通路甜菜素生物合成、吲哚生物碱生物合成和加压素调节水的重吸收随着微贮棉秆比例升高而显著增加(P<0.05)。【结论】饲喂50%微贮棉秆在提高日增重的同时对湖羊瘤胃微生物菌群结构与功能影响较小。在生产实践中,微贮秸秆添加量应低于50%。

厌氧真菌和甲烷菌共培养的研究进展

厌氧真菌是自然界中降解植物纤维素类物质最高效的微生物之一。近年来,大量厌氧真菌和甲烷菌共培养菌株被分离。共培养中,甲烷菌通过对厌氧真菌代谢产物的利用显著提高厌氧真菌对木质纤维素的降解;厌氧真菌通过为甲烷菌提供能量和营养物质使甲烷菌快速生成大量甲烷。全面深入地了解共培养中两者的互作关系以及共培养降解木质纤维素产甲烷的特性,将有助于研究对木质纤维素降解以及甲烷生成的调控。因此,本文主要综述了共培养的分离鉴定、多样性、互作关系以及对木质纤维素的降解。

Gut microbiome colonization and development in neonatal ruminants: Strategies, prospects, and opportunities

Colonization and development of the gut microbiome is a crucial consideration for optimizing the health and performance of livestock animals. This is mainly attributed to the fact that dietary and management practices greatly influence the gut microbiota, subsequently leading to changes in nutrient utilization and immune response. A favorable microbiome can be implanted through dietary or management in-terventions of livestock animals, especially during early life. In this review, we explore all the possible factors (for example gestation, colostrum, and milk feeding, drinking water, starter feed, inoculation from healthy animals, prebiotics/probiotics, weaning time, essential oil and transgenesis), which can influence rumen microbiome colonization and development. We discuss the advantages and disadvantages of potential strategies used to manipulate gut development and microbial colonization to improve the production and health of newborn calves at an early age when they are most susceptible to enteric disease. Moreover, we provide insights into possible interventions and their potential effects on rumen development and microbiota establishment. Prospects of latest techniques like transgenesis and host genetics have also been discussed regarding their potential role in modulation of rumen microbiome and subsequent effects on gut development and performance in neonatal ruminants.

猪源乳杆菌Lactobacillus reuteri strain DSM和Lactobacillus taiwanensis strain BCRC对肠道炎症的影响

【目的】旨在以葡聚糖硫酸钠(DSS)诱导小鼠肠炎模型研究猪源乳杆菌Lactobacillus reuteri strain DSM(LR)和Lactobacillus taiwanensis strain BCRC(LT)对肠道炎症的影响。【方法】选取24只8周龄体重相近的C57BL/6J雄性小鼠,随机分为4组(每组6个重复):正常对照组、DSS组、LR菌处理组(DSS+LR)和LT菌处理组(DSS+LT)。适应期7 d,试验期16 d,随后处死小鼠并采样。【结果】与正常对照组相比,DSS组小鼠体重、结肠长度显著降低,疾病活动指数(DAI)显著增加(P<0.05);与DSS组相比,LR、LT菌处理组小鼠体重下降的程度并没有显著缓解,但是,其DAI评分显著降低;并且,LT菌处理组结肠长度显著增加(P<0.05)。正常对照组和LT菌处理组结肠组织病理学评分显著低于DSS组和LR菌处理组(P<0.05)。结肠髓过氧化物酶(MPO)活性在正常对照组和LT菌处理组也显著低于DSS组(P<0.05),在LR菌处理组与DSS组无显著差异(P>0.05)。与正常对照组相比,DSS组结肠和血浆的促炎因子TNF-α、IL-1β、IL-6水平显著升高,而抗炎因子IL-10水平显著降低;相比于DSS组,LT菌处理组显著降低了结肠和血浆的TNF-α、IL-1β、IL-6水平,升高了IL-10水平;LR菌处理组显著降低了结肠TNF-α水平、血浆IL-1β水平,升高IL-10水平(P<0.05)。免疫组化和ELISA结果显示,相比于正常对照组,DSS组降低了结肠紧密连接蛋白Claudin-1、Occludin、ZO-1和黏蛋白Muc-2表达水平,而LR或LT菌的处理则上调了DSS影响下的屏障相关蛋白表达。16S rDNA测序技术检测结肠菌群组成,PCoA分析得出,正常对照组和DSS组显著分离,LT处理组菌群组成更接近于正常对照组。门水平上,变形菌门丰度在正常对照组显著低于DSS组(P<0.05);与DSS组相比,LT菌处理组也能降低变形菌门丰度,但未达显著水平(P>0.05)。属水平上,Muribaculaceae_norank属在正常对照组显著高于其他3组,Bacteroides在正常对照组显著低于其他3组(P<0.05);与DSS组相比,LR或LT菌的处理增加了Muribaculaceae_norank丰度,降低了Bacteroides菌丰度,但差异不显著(P>0.05)。【结论】乳杆菌Lactobacillus reuteri和Lactobacillus taiwanensis能增强肠上皮屏障,对DSS引起的肠道炎症起到了一定的缓解作用;其中,Lactobacillus taiwanensis对肠上皮屏障的保护效果更为明显。

The uniqueness and superiority of energy utilization in yaks compared with cattle in the highlands:A review

Yaks living on the Qinghai-Tibetan Plateau for a long time have evolved a series of mechanisms to adapt to the unique geographical environment and climate characteristics of the plateau.Compared with other ruminants,yaks have higher energy utilization and metabolic efficiency.This paper presents possible mechanisms responsible for the efficient energy utilization,absorption and metabolism resulting from the unique evolutionary process of yaks.It is hoped that the information discussed in this review will give a better insight into the uniqueness and superiority of yaks in regards to energy metabolism and utilization compared with cattle and open new avenues for the targeted regulation of energy utilization pathways of other ruminants.

Characterization of a novel bifunctional enzyme from buffalo rumen metagenome and its effect on in vitro ruminal fermentation and microbial community composition

To efficiently use lignocellulosic materials in ruminants,it is crucial to explore effective enzymes,especially bifunctional enzymes.In this study,a novel stable bifunctional cellulase-xylanase protein from buffalo rumen metagenome was expressed and characterized,CelXyn2.The enzyme displayed optimal activity at pH 6.0 and 45℃.The residual endoglucanase and xylanase activities were 90.6%and 86.4%after a 60-min pre-incubation at 55℃.Hydrolysis of rice straw,wheat straw,sheepgrass and sugar beet pulp by CelXyn2 showed its ability to degrade both cellulose and hemicellulose polymers.Treatment with CelXyn2 improved the hydrolysis of agricultural residues with an evident increase in production of total gas,lactate and volatile fatty acids.The results of 16S rRNA and real-time PCR showed that the effect on in vitro ruminal microbial community depended on fermenta-tion substrates.This study demonstrated that CelXyn2 could strengthen lignocellulose hydrolysis and in vitro ruminal fermentation.These characteristics of CelXyn2 distinguish it as a promising candidate for agricultural application.