Investigation of Interspecies Interactions within Marine Micromonosporaceae Using an Improved Co-Culture Approach

With respect to bacterial natural products, a significant outcome of the genomic era was that the biosynthetic potential in many microorganisms surpassed the number of compounds isolated under standard laboratory growth conditions, particularly among certain members in the phylum Actinobacteria. Our group, as well as others, investigated interspecies interactions, via co-culture, as a technique to coax bacteria to produce novel natural products. While co-culture provides new opportunities, challenges exist and questions surrounding these methods remain unanswered. In marine bacteria, for example, how prevalent are interspecies interactions and how commonly do interactions result in novel natural products? In an attempt to begin to answer basic questions surrounding co-culture of marine microorganisms, we have tested both antibiotic activity-based and LC/MS-based methods to evaluate Micromonosporaceae secondary metabolite production in co-culture. Overall, our investigation of 65 Micromonosporaceae led to the identification of 12 Micromonosporaceae across three genera that produced unique metabolites in co-culture. Our results suggest that interspecies interactions were prevalent between marine Micromonosporaceae and marine mycolic acid-containing bacteria. Furthermore, our approach highlights a sensitive and rapid method for investigating interspecies interactions in search of novel antibiotics, secondary metabolites, and genes.     

基于UPLC-QTOF/MS的小麦发芽代谢组学分析方法

为探究不同提取方法对发芽小麦代谢物提取效果的影响,本研究建立了基于超高效液相色谱-四级杆飞行时间质谱(UPLC-QTOF/MS)的发芽小麦非靶向代谢组学样品前处理方法和分析方法,以发芽2 d周麦26籽粒为材料,设计提取溶剂、提取方式、提取时间3个因素在3个水平上L9(34)正交试验,并通过主成分分析和聚类分析确定提取效果最佳的组合。以80%乙腈(0.1%甲酸)震摇提取30 min可检测出1609个代谢峰,说明提取溶剂对代谢物提取效果起主要作用。共鉴定出92种小麦代谢物。     

基于UHPLC-QTOF-MS代谢组学方法筛选奶牛不同妊娠状态时的候选生物标志物

旨在筛选人工授精后第17天奶牛在不同妊娠状态时的候选生物标志物。本研究以宁夏某奶牛场体重为（550±50） kg,体况评分相近的经产（2~3胎次）健康荷斯坦奶牛为试验对象。同期发情后,在人工授精后第17天晨饲前对奶牛进行尾静脉采血,后期采用计步器和B超仪诊断出奶牛的妊娠状态。根据诊断结果将奶牛分为妊娠组（A组,n=12）和未妊娠返情组（B组,n=24）,将这两组血样进行代谢轮廓及代谢物变化分析。主成分分析（PCA）和正交偏最小二乘法-判别分析（OPLS-DA）结果显示,A、B两组血浆代谢轮廓均发生了明显变化,A、B两组之间共检测出8种ROC （Receiver operating characteristic curve）曲线下面积（area under the ROC curve,AUC）＞0.8的差异代谢物。丙氨酸-亮氨酸、甘氨酸、脯氨酸、L-正亮氨酸、DL-苯丙氨酸、肌氨酸、吡咯-2-羧酸和缬氨酸-蛋氨酸有望成为识别妊娠组（A组）和未妊娠返情组（B组）的差异代谢物。综上表明,血浆中的这8种代谢物有望成为奶牛妊娠识别阶段潜在的生物标志物,为配种早期妊娠识别提供新思路。     

Taurine alone or in combination with fish protein hydrolysate affects growth performance,taurine transport and metabolism in juvenile turbot (Scophthalmus maximus L.)

The study was conducted to investigate the effects of taurine (Tau) alone or in combination with fish protein hydrolysate (FPH) on growth performance, the expression of Tau transporter (TauT) and metabolic profile in juvenile turbot. FM, FPH0, FPH0+T, FPH10 and FPH10+T diets, respectively, contained 300, 150, 150, 80, and 80 g/kg fishmeal. FPH10 and FPH10+T diets contained 62 g/kg FPH. FPH0+T and FPH10+T diets were, respectively, prepared by supplementing the FPH0 and FPH10 diet formulations with 8 g/kg Tau. Specific growth rate was the highest in FM group and the lowest in FPH10 group. TauT mRNA levels in fish fed Tau supplemented diets were significantly lower than that in Tau unsupplemented diets. NMR‐based metabolomics analysis showed that Tau contents in liver of FPH0+T and FPH10+T were significantly higher than that of FM, FPH0 and FPH10. In muscle, Tau contents were significantly decreased in the FPH10+T versus FPH0 and the FPH10+T versus FPH10 comparisons. In conclusion, 62 g/kg FPH to replace fishmeal may not affect Tau synthesis, transport and metabolism. However, Tau supplemented alone or in combination with a certain level of FPH could reduce the requirement for Tau synthesis and transport and increased Tau levels in muscle and liver.     

基于UHPLC-Q-TOF/MS的不同产地普洱生茶化学成分差异研究

为了更加全面地了解产地对普洱生茶品质与化学成分的影响,本研究选取来自临沧市、普洱市、西双版纳州三大产区12个茶山(自然村)的普洱生茶样品,采用超高效液相色谱-四级杆-飞行时间质谱(UHPLC-Q-TOF/MS)对不同产地普洱生茶的非挥发性代谢物表型进行了研究。结果表明:不同产地普洱生茶的化学成分在含量上具有较大差异,且具有明显的产地特征。采用主成分分析,可以对来自西双版纳自治州(包括勐腊县、勐海县、景洪市)、普洱市、临沧市3个地级行政区的普洱生茶进行有效区分,也可以对来自普洱茶产区的东南、西南、西北3个区域的普洱生茶进行有效区分。进一步鉴定了普洱生茶中79种主要成分,并对其在12个茶山(自然村)的普洱生茶中的含量分布,以及与不同产地普洱生茶滋味品质的关系进行了分析。本研究表明,基于UHPLC-Q-TOF/MS的茶叶非挥发性化学成分轮廓可以作为普洱生茶产地判别的依据。     

基于血浆非靶代谢组学探究急性笼养应激对蛋鸭代谢的影响

随着环保政策改革和养殖业的集约化、规模化发展,蛋鸭叠层式笼养模式的应用越来越广泛,笼养应激对蛋鸭的影响逐渐受到重视。本研究旨在利用非靶向代谢组学分析蛋鸭平养与笼养模式下血浆代谢物的差异。随机挑选平养组(floor-water rearing system, FWR组)和叠层式笼养组(cage-rearing system, CR组)蛋鸭各10只,跖骨静脉采血,通过高效液相色谱-质谱(HPLC-HRMS)非靶向代谢组学技术获得20个在不同饲养模式下的血浆代谢物图谱,经T检验等分析,取差异倍数(Fold-change)ratio≥2或ratio≤1/2、变量重要性(variable importance in projection, VIP)>1、q值<0.05为筛选条件得到显著差异代谢物并作通路富集分析。结果共筛选到差异代谢物30种,其中上调表达的差异代谢物14种(P<0.05),下调表达的差异代谢物16种(P<0.05)。差异代谢物种类主要有甘油磷脂(glycerophospholipids, 16种)、脂肪酰基(fatty acyls, 4种)、羧酸及其衍生物...