基于LC-MS技术的代谢组学方法研究吡虫啉对工蜂代谢的影响

试验利用液相色谱-质谱联用技术（liquid chromatograph mass spectrometer,LC-MS）对出房的1日龄工蜂进行代谢组学分析,以明确亚致死剂量的吡虫啉对蜜蜂生长发育期代谢的影响。采用蜜蜂幼虫人工室内饲养技术,选取意大利蜜蜂（Apis mellifera ligustica）为研究对象,用含0.00001μg/μL亚致死剂量吡虫啉的日粮饲喂1日龄幼虫直至排便。记录蜜蜂的羽化率、初生重、出房时间;测量蜜蜂血淋巴的超氧化物歧化酶（super oxide dismutase,SOD）、过氧化氢酶（catalase,CAT）、过氧化物酶（peroxidase,POD）活力;采用LC-MS方法检测亚致死剂量浓度吡虫啉处理蜜蜂羽化出房时的代谢物差异,对检测数据进行模式识别分析,筛选差异代谢物。结果表明,亚致死剂量的吡虫啉引起蜜蜂出房时间显著延长（P<0.05）;使蜜蜂血淋巴的SOD和POD活性显著升高（P<0.05）;采用主成分分析（principal component analysis,PCA）和偏最小二乘法判别方法（supervised partial leastsquares-discriminant analysis,PLS-DA）分析代谢组数据,结果显示试验组和对照组明显分离。试验共鉴定出18个蜜蜂差异代谢物。本结果可对进一步研究吡虫啉胁迫蜜蜂代谢物的代谢规律、阐明蜜蜂解毒机制提供理论依据。     

家畜肠道微生物代谢组学研究进展

家畜肠道微生物群落之间、微生物与宿主之间不断相互作用,形成动物肠道内复杂的微生物生态环境。肠道微生物及其代谢产物对维持家畜正常生长发育、营养代谢、繁殖性能等方面有着重要作用。微生物-脑-肠轴是中枢神经系统和肠道微生物之间的双向调节渠道。利用代谢组学对微生物代谢物进行检测能对微生物多样性分析数据进行补充,从而进一步揭示肠道微生物与宿主之间相互作用关系。该文综述了家畜肠道微生物代谢物的种类、代谢组学在肠道微生物研究中的应用以及肠道微生物代谢物对家畜动物的影响等内容,有助于了解微生物代谢物与家畜之间的相互作用以及研究的现状。     

微生物代谢组学及其在饲料产品开发中的应用

代谢组学是继基因组学、转录组学、蛋白质组学发展起来的一门新的学科,微生物代谢组学作为其中的一个重要分支.从微生物产品开发到代谢工程的应用,微生物代谢组学已成为研究热点.本文主要综述了微生物代谢组学的一般研究流程,即试验设计、样本采集、数据获取、数据处理与生物学解释,及其在动物饲料添加剂和微生物发酵饲料开发中的前景.     

杜仲叶影响绵羊血清代谢组学的研究

为探索杜仲叶（Eucommia ulmoides leaves,EUL）对绵羊营养代谢和生理功能的影响,本研究选取25~30 kg、70~80日龄的湖羊30只,随机分为3组,每组10只,分别为对照组（无杜仲叶日粮组,CTL）、低剂量添加组（10%杜仲叶日粮组,EUL1）、高剂量添加组（20%杜仲叶日粮组,EUL2）。预试期15 d,正试期90 d。试验结束时,通过静脉采血,离心后分别取血浆和血清,血浆用于生化分析,血清用于代谢组学分析。结果显示,饲喂杜仲叶后,EUL1和EUL2组绵羊血浆中葡萄糖、非酯化脂肪酸、高密度脂蛋白和极低密度脂蛋白均升高,EUL2组与CTL组差异显著或极显著（P<0.05;P<0.01）,EUL1组与CTL组差异不显著（P>0.05）;EUL1和EUL2组尿素较CTL组极显著降低（P<0.01）,总胆固醇水平显著降低（P<0.05）,EUL1和EUL2组间差异不显著（P>0.05）。血清样品采用液相色谱-质谱联用仪（LC-MS）进行检测,在正模式下得到593个代谢物,在负模式下得到1 570个代谢物。通过主成分分析（PCA）、偏最小二乘判别分析（PLS-DA）及正交偏最小二乘判别分析（OPLS-DA）发现,各试验组间的差异代谢物较多,能将其得分图明显区分。进一步对变量重要性因子大于1.0（VIP>1.0）的代谢物进行t检验后发现,各组之间有24种特征代谢物差异显著（P<0.05）。这些差异代谢物涉及体内葡萄糖、脂肪酸和氨基酸等的代谢,有的代谢物还与动物的免疫机能相关。杜仲叶饲料对绵羊的营养代谢、生理状态和免疫状况可产生明显的影响,代谢组学可有针对性地分析代谢物发生的步骤及生理作用,为阐明杜仲叶影响营养代谢的机制提供参考。     

基于组学技术研究反刍动物瘤胃微生物及其代谢功能的进展

瘤胃微生物对反刍动物的生产效率和健康状况以及温室气体的排放起着至关重要的作用,而采用不同的研究方法和手段揭示瘤胃生态系统中微生物群落的结构组成和代谢功能是该研究领域的热点和重点。本文综述了采用多组学(宏基因组学、宏转录组学、宏蛋白质组学和代谢组学)技术,结合不断发展的仪器分析和生物信息技术,研究反刍动物瘤胃微生物组成、基因组功能及代谢方面的最新进展,旨在为进一步调控瘤胃微生物以提高反刍动物生产和减轻环境污染提供新的技术手段和理论基础。     

岩藻多糖对断奶羔羊瘤胃微生物区系及代谢组学的影响

本试验旨在研究岩藻多糖对断奶羔羊瘤胃微生物区系及代谢组学的影响。选用60只60日龄平均体重为（12.5±0.5） kg的健康断奶羔羊（川中黑山羊）,随机分为4组,每组15只。对照组（CON组）饲喂基础饲粮,试验组在基础饲粮中分别添加0.1%(F1组)、0.3%(F2组)和0.5%(F3组)的岩藻多糖。预试期7 d,正试期30 d。结果表明：1）与CON组相比,F2、F3组的末重、平均日增重和平均日采食量显著升高（P<0.05）,F1、F2、F3组的料重比显著降低（P <0.05）。2）与CON组相比,F1、F2、F3组的瘤胃pH和乙酸、丁酸、异丁酸含量及乙酸/丙酸显著升高（P<0.05）,F2、F3组的瘤胃丙酸含量显著升高（P<0.05）。3）在门水平上,与CON组相比,F1、F2、F3组的瘤胃中疣微菌门（Verrucomicrobia）的相对丰度显著升高（P<0.05）;F1、F3组的瘤胃中拟杆菌门（Bacteroidetes）的相对丰度显著升高（P<0.05）,放线菌门（Actinobacteria）、广古菌门（Euryarchaeota）、螺旋...     

脂质代谢组学的研究进展

脂质是动物体内重要的生物大分子之一,具有多种重要生物学功能。脂质代谢组学是新兴起的一门研究脂质代谢调控在各种生命现象中作用机制的新学科。它通过从系统水平上研究生物体内的脂质,揭示脂质分子及与其他生物分子间的相互作用。脂质代谢组学在营养学、功能基因组学等相关学科中的应用已有许多报道。本文主要介绍了脂质的分类与代谢、脂质代谢组学的概念、研究方法、研究现状及其应用研究进展等。     

基于代谢组学的乳汁中代谢物研究进展北大核心CSCD

乳汁是为新生哺乳动物的生长发育提供必需营养的最佳食物。除了为新生哺乳动物提供营养外,还含有大量小分子代谢物,这些代谢物在母体和新生哺乳动物之间形成了复杂的联系。在对乳汁的研究中应用代谢组学分析可以挖掘其中小分子物质的变化情况,结合生物信息学分析能够将关键代谢标志物呈现,并预测营养素等对乳汁代谢组影响的作用机制。本文主要总结了人类乳汁和奶牛乳汁分析中应用到代谢组学技术的文献,并结合其他关于乳汁或代谢组学的研究,阐述了乳汁中部分低丰度差异代谢物的变化及其影响因素,剖析了未来在乳汁分析中的潜在方向,以期为揭示乳汁代谢组的生理机制提供新思路。     

基于GC/TOF-MS技术对流感感染小鼠粪便代谢组学的研究

为了探讨小鼠感染流感病毒后粪便代谢物的变化情况,寻找潜在生物标志物及相关的代谢通路,本研究运用气相色谱-飞行时间质谱联用技术(GC/TOF-MS)检测对照组和病毒组小鼠的粪便。结果显示:在正交偏最小二乘法分析(OPLS)模型中,对照组与病毒组的得分图呈现明显差异,病毒组中一些代谢物发生了显著性的改变。α-酮戊二酸、甲硫氨酸、果糖-6-磷酸、苯丙氨酸、草酸、乳糖、棕榈酸、正缬氨酸、腺嘌呤、胆固醇、葡萄糖-6-磷酸、油酸、甘油、十五烷酸、β-丙氨酸等代谢物显著降低,而L-苹果酸、天冬氨酸、乳酸等代谢物显著上升。结论:病毒组与对照组之间存在脂类代谢、氨基酸代谢及糖代谢的差异,代谢组学的研究可以为流感病毒引起的肠道损伤机制提供新的依据。     

The effects of acute lipopolysaccharide challenge on dairy goat liver metabolism assessed with 1HNMR metabonomics

We investigated the mechanisms mediating hepatic metabolic responses to an acute lipopolysaccharide (LPS) challenge in goats. Guanzhong dairy goats (15) were randomly divided into three groups: control (CTL, saline, 0.2 ml/kg BW), lower dose LPS (LPS‐L, 20 μg/kg BW) and higher dose LPS (LPS‐H, 40 μg/kg BW). All injections were administered intraperitoneally twice with a 24‐h interval. Forty‐eight hours after the first injection, blood samples were collected to extract plasma for biochemical analysis, and liver tissues were biopsied and stored in liquid nitrogen for metabonomics analysis. We found that plasma levels of alanine aminotransferase, aspartate aminotransferase and total bilirubin increased (p < 0.05) in both LPS‐treated groups, whereas plasma triglyceride, cholesterol, very low‐density lipoprotein, low‐density lipoprotein, high‐density lipoprotein, total protein and albumin levels markedly decreased (p < 0.05). The increased activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), levels of tumour necrosis factor α (TNF‐α), interleukin (IL)‐1β, IL‐6 and IL‐8 indicated hepatic injury and metabolic dysfunction in some degree. Using proton nuclear magnetic resonance (¹H‐NMR) metabonomics and the Chenomx NMR suite database, 69 metabolites were detected and identified. Metabolic differences among the groups were determined with pattern recognition analyses using principal component analysis and supervised projection to latent structures discriminant analysis. Pattern recognition analysis distinguished and clustered the metabolite variables from the three groups, finding nine of 69 metabolites that differed significantly between two of the three groups: six from the LPS‐L or LPS‐H groups differed from CTL and three differed between LPS‐L and LPS‐H groups. These altered metabolites were closely connected with glucose, lipid and amino acid metabolic pathways in hepatocytes. Based on an analysis of these metabolites and their relevant pathways, the mechanisms and degree of hepatic injury were deduced. Therefore, the metabolic profile was used effectively to detect characteristic hepatic metabolites, discriminate metabolic changes induced by LPS, clarify the mechanisms for the resulting metabolic dysfunctions and provide efficient information to diagnose liver injury.     

微生物脂肪酶概况及其动物营养效应的研究进展

微生物脂肪酶是工业用脂肪酶的重要来源,是生物技术和有机化学应用中广泛使用的酶类之一。文章从微生物脂肪酶来源、酶学特性、在动物消化道中的营养效应和作用机理方面进行综述,主要介绍国内外关于脂肪酶应用在动物饲料方面所取得的一些研究成果,为微生物脂肪酶在饲料中的开发与应用提供参考。     

The crucial role of lysine in the hepatic metabolism of growing Holstein dairy heifers as revealed by LC-MS-based untargeted metabolomics

The objective of this experiment was to evaluate the effect of supplementing rumen-protected Lys based on a Lys-deficient diet on liver metabolism in growing Holstein heifers. The experiment was conducted for 3 months with 36 Holstein heifers (initial body weight: 200 ± 9.0 kg; 7-month-old). Heifers were randomly assigned to 2 diets based on corn, soybean meal, alfalfa hay, and wheat bran: control, Lys-deficient diet (LD; 0.66% Lys in diet), and Lys-adequate diet (LA; 1.00% Lys in diet). The results showed no difference in growth performance between the 2 groups (P > 0.05). However, there was a clear trend of increasing feed conversion rate with Lys supplementation (0.05 < P < 0.01). The serum urea nitrogen concentration was significantly decreased, and the aspartate aminotransferase-to-alanine aminotransferase ratio was significantly decreased by Lys supplementation (P < 0.05). Moreover, growing heifers fed a Lys-adequate diet had lower levels of urine nitrogen excretion and higher levels of the biological value of nitrogen (P < 0.05). Metabolomic analysis revealed that 5 types of phosphatidylcholine and 3 types of ceramide were significantly increased and enriched in sphingolipid metabolism and glycerophospholipid metabolism (P < 0.05). His, Leu, and Asp levels were significantly decreased in the liver following Lys supplementation (P < 0.05). In conclusion, Lys supplementation may promote the synthesis of body tissue proteins, as evidenced by significantly decreased amino acids in the liver and urine N excretion, it also improves hepatic lipid metabolism by providing lipoprotein precursors.     

The potential for mitigation of methane emissions in ruminants through the application of metagenomics,metabolomics, and other -OMICS technologies

Ruminant supply chains contribute 5.7 gigatons of CO_2-eq per annum, which represents approximately 80% of the livestock sector emissions. One of the largest sources of emission in the ruminant sector is methane (CH₄), accounting for approximately 40% of the sectors total emissions. With climate change being a growing concern, emphasis is being put on reducing greenhouse gas emissions, including those from ruminant production. Various genetic and environmental factors influence cattle CH₄ production, such as breed, genetic makeup, diet, management practices, and physiological status of the host. The influence of genetic variability on CH₄ yield in ruminants indicates that genomic selection for reduced CH₄ emissions is possible. Although the microbiology of CH₄ production has been studied, further research is needed to identify key differences in the host and microbiome genomes and how they interact with one another. The advancement of “-omics” technologies, such as metabolomics and metagenomics, may provide valuable information in this regard. Improved understanding of genetic mechanisms associated with CH₄ production and the interaction between the microbiome profile and host genetics will increase the rate of genetic progress for reduced CH₄ emissions. Through a systems biology approach, various “-omics” technologies can be combined to unravel genomic regions and genetic markers associated with CH₄ production, which can then be used in selective breeding programs. This comprehensive review discusses current challenges in applying genomic selection for reduced CH₄ emissions, and the potential for “-omics” technologies, especially metabolomics and metagenomics, to minimize such challenges. The integration and evaluation of different levels of biological information using a systems biology approach is also discussed, which can assist in understanding the underlying genetic mechanisms and biology of CH₄ production traits in ruminants and aid in reducing agriculture’s overall environmental footprint.     

直接饲喂微生物在反刍动物生产上的应用

直接饲喂微生物（DFM）能提高育肥牛日增重和饲料利用率，提高奶牛泌乳量，改善幼龄反刍动物的健康状况和生产成绩。然而，DFM的作用很不稳定，其作用模式不明。文中主要介绍了DFM对反刍动物健康和生产性能的影响，讨论了其潜在的作用机理。并对有待于继续研究的问题进行了探讨。     

微生物饲料添加剂在畜禽生产中的应用

微生物饲料添加剂无污染、无残留、安全无毒，在畜禽生产中的应用日益广泛。在使用微生物饲料添加剂的过程中，只有全面了解微生物饲料的作用机理和产品特性，正确对待其作用效果，才能使其发挥更显著的作用。     

瘤胃微生物多样性与分析技术的研究进展

瘤胃微生物区系是一个组成和功能极其复杂的体系,微生物的多样性决定着动物机体健康和生产性能。本文就瘤胃微生物多样性的研究进展和瘤胃微生物分类定量的技术手段以及益生菌对瘤胃微生物的作用作一综述。     

松嫩草甸寸草苔群落土壤微生物量磷的初步研究

松嫩草甸寸草苔群落是羊草群落轻微碱化演替的次生群落。对该群落土壤微生物量磷季节变化、垂直分布以及与土壤因子的关系进行初步研究。结果表明,0~10和20~30 cm土层微生物量磷的季节变化趋势呈明显的单峰曲线,峰值出现在8月;10~20 cm土层微生物量磷的季节变化不明显;表层(0~10 cm)土壤微生物量磷含量高于10~20和20~30 cm土层的含量。通径分析结果表明,速效氮、全磷、缓效钾以及土壤pH值和土壤温度对土壤微生物量磷的直接影响较大,而土壤速效磷主要是通过其他土壤因子间接影响土壤微生物量磷;紧实的土壤条件对土壤微生物代谢活动具有明显的限制作用,而土壤盐化状况对微生物代谢的限制作用不大。逐步回归分析结果表明,上层(0~20 cm)土壤全磷含量是促进土壤微生物量磷累积的重要因素,而下层(20~30 cm)土壤微生物对磷素的累积主要受到土壤可利用磷素的间接影响。     

水产养殖用复合微生物粉剂中菌种的分子生物学鉴定

对两种市售水产养殖用复合微生物粉剂中微生物菌种进行鉴定。利用平板分离技术对粉剂中的微生物进行分离纯化,进行了形态学观察、16S rDNA和26S rDNA序列分析,并构建系统发育树。结果表明,两种微生物粉剂中菌群结构存在一定差异,S1粉剂中鉴定出5株芽孢杆菌和2株酵母菌,分别为枯草芽孢杆菌、地衣芽孢杆菌或索诺拉沙漠芽孢杆菌、葡萄牙棒孢酵母和库德里阿兹威毕赤酵母;S2粉剂中鉴定出3株芽孢杆菌、1株放线菌、2株酵母菌和3株肠球菌,分别为解木糖赖氨酸芽孢杆菌、环状芽孢杆菌、地衣芽孢杆菌或索诺拉沙漠芽孢杆菌、芬氏纤维微菌、酿酒酵母、库德里阿兹威毕赤酵母、鹑鸡肠球菌、屎肠球菌或乳酸肠球菌。明确水产养殖用复合微生物粉剂的菌群结构为同类产品的安全性及有效性评估提供理论基础。