Metabolomics Tools Assisting Classic Screening Methods in Discovering New Antibiotics from Mangrove Actinomycetia in Leizhou Peninsula

Mangrove actinomycetia are considered one of the promising sources for discovering novel biologically active compounds. Traditional bioactivity- and/or taxonomy-based methods are inefficient and usually result in the re-discovery of known metabolites. Thus, improving selection efficiency among strain candidates is of interest especially in the early stage of the antibiotic discovery program. In this study, an integrated strategy of combining phylogenetic data and bioactivity tests with a metabolomics-based dereplication approach was applied to fast track the selection process. A total of 521 actinomycetial strains affiliated to 40 genera in 23 families were isolated from 13 different mangrove soil samples by the culture-dependent method. A total of 179 strains affiliated to 40 different genera with a unique colony morphology were selected to evaluate antibacterial activity against 12 indicator bacteria. Of the 179 tested isolates, 47 showed activities against at least one of the tested pathogens. Analysis of 23 out of 47 active isolates using UPLC-HRMS-PCA revealed six outliers. Further analysis using the OPLS-DA model identified five compounds from two outliers contributing to the bioactivity against drug-sensitive A. baumannii. Molecular networking was used to determine the relationship of significant metabolites in six outliers and to find their potentially new congeners. Finally, two Streptomyces strains (M22, H37) producing potentially new compounds were rapidly prioritized on the basis of their distinct chemistry profiles, dereplication results, and antibacterial activities, as well as taxonomical information. Two new trioxacarcins with keto-reduced trioxacarcinose B, gutingimycin B (16) and trioxacarcin G (20), together with known gutingimycin (12), were isolated from the scale-up fermentation broth of Streptomyces sp. M22. Our study demonstrated that metabolomics tools could greatly assist classic antibiotic discovery methods in strain prioritization to improve efficiency in discovering novel antibiotics from those highly productive and rich diversity ecosystems.     

Fate of Endosulfan in Cotton Soil under Sub-tropical Conditions of Northern India

Terrestrial field studies were conducted with endosulfan during the 1989–90 Kharif season in bare cotton soil, to investigate the fate of endosulfan and its downward movement under sub-tropical conditions of northern India. Field experiments consisted of spray application of endosulfan at 875 g ha^-1 42 and 63 days after the assumed date of sowing in two separate treatments. Soil samples drawn periodically from different depths were analysed by GC-ECD (Ni⁶³) for endosulfan and its breakdown products. Dissipation of the total endosulfan residues occurred to an extent of 92–97% in the first four-week period and by about 99% in 238 days in two distinct phases in first-order kinetics. Residue half life (T_1/2) varied from 39 to 42 days. The parent compound metabolized to endosulfan-diol and endosulfan sulfate. Endosulfan-diol remained confined in the upper 5-cm layer and dissipated completely in 28 days whereas endosulfan sulfate was first detectable seven days after treatment and persisted until the end of the experiment, remaining confined in the upper 0–10 cm soil layer. The β-isomer also did not leach down beyond 10 cm depth. © 1997 SCI.     

Differences in metabolism of triploid and diploid Salmo trutta f. lacustris under acclimation conditions and after exposure to stress situations

Differences in metabolism (enzyme activities, metabolites) between diploid and triploid Salmo trutta f. lacustris were investigated under acclimation and stress conditions. Under acclimation conditions enzyme activities differed for 35% of the 27 investigated key regulatory enzymes and temperature optima for 23%. Muscle and liver metabolites related to energy metabolism and diagnostic indices of blood serum were similar, with exception of acetyl‐CoA being increased in triploids. Metabolic rate was lower and gill ventilation rate higher in triploids in comparison with diploids. During the tested stress situations (24 hr endurance swimming, 3 hr exposure to hypoxia in water with 32% oxygen saturation) muscle and liver glycogen decreased and serum and muscle lactate increased in both ploidy levels. Specifically, for triploids muscle adenylate energy charge and phosphocreatine levels decreased after endurance swimming and muscle and liver adenylate energy charge after exposure to hypoxia. Acetyl‐CoA increased in triploids during both stress situations. In summary, there existed differences in metabolism between the two ploidy levels and the energy metabolism of triploids was less balanced under stress.     

超高效液相色谱-串联质谱法测定鱼粉中喹噁啉类药物及其主要代谢物的残留量

文章建立了超高效液相色谱-串联质谱法同时检测鱼粉中5种喹噁啉类药物及其2种主要代谢物残留量的方法。样品经乙腈-乙酸乙酯(1∶1,V∶V)和1 mol·L^–1盐酸分步提取,盐酸提取液进一步用乙酸乙酯反萃取,有机相经浓缩后,均用乙腈复溶,用PRiME HLB通过性固相萃取柱净化处理。样品以乙腈和0.1%甲酸水溶液为流动相,经Phenomenex Kinetex C18柱梯度洗脱分离,质谱采取正离子多反应监测模式进行检测。代谢物采用内标法定量,原药采用基质匹配外标法定量。结果表明,7种化合物在对应的浓度范围内线性关系良好(R≥0.994),2种代谢产物检测限为2μg·kg–1,定量限为5μg·kg^–1;5种喹噁啉类药物检测限为1~10μg·kg^–1;定量限为2~20μg·kg^–1。在高、中、低3种添加浓度下7种化合物的平均回收率为64.4%~102.2%,相对标准偏差为3.2%~10.2%。方法灵敏度高、精密度好,能同时测定鱼粉中的喹噁啉类药物及其主要代谢物。     

桂西北喀斯特地区不同土地利用类型土壤抗蚀性研究

土壤抗蚀性是反映土壤抵抗侵蚀能力的重要参数之一,是土壤侵蚀研究的重要内容。本文选取土壤有机质、水稳性团聚体、团聚体结构破坏率、团聚状况、团聚度、分散率和<0.05 mm粉黏粒含量等7个指标,通过单因素方差分析及主成分分析,探讨了桂西北喀斯特地区5种不同土地利用类型土壤抗蚀性的差异。结果表明:原生林和次生林土壤有机质含量显著(P<0.05)高于撂荒地、坡耕地和人工林,撂荒地土壤有机质含量较坡耕地和人工林高,但差异不显著。原生林、次生林及撂荒地土壤>0.25 mm水稳性团聚体总量及团聚状况显著高于坡耕地及人工林,但其土壤团聚体结构破坏率及分散率显著低于坡耕地;人工林土壤团聚体结构破坏率显著高于次生林,但与原生林、撂荒地和坡耕地差异不显著;人工林土壤分散率则与坡耕地类似,显著低于原生林、次生林及撂荒地;原生林、次生林土壤团聚度与撂荒地、坡耕地差异不显著,但显著高于人工林;次生林、撂荒地及人工林<0.05 mm粉黏粒含量与原生林、坡耕地差异不显著,但坡耕地土壤<0.05 mm粉黏粒含量显著高于原生林。由主成分分析综合评分得到土壤抗蚀性强弱顺序为:原生林>次生林>撂荒地>坡耕地>人工林。因此,喀斯特地区人为干扰严重降低了土壤的抗蚀性,耕地通过撂荒方式能够提高土壤抗蚀性。     

安格斯妊娠母牛异食行为的营养代谢异常及其与肠道菌群相关性研究

牛异食行为(PICA)是以舔食、啃咬一些无营养价值异物为特征的一种异常行为,大多研究认为,PICA是因某些营养素紊乱和生活环境受限所致.业已证明,肠道菌群与宿主营养代谢关系密切,肠道菌群-肠-脑(micro-biota-gut-brain,MGB)轴与动物行为之间的相关性研究日益被揭示.但PICA牛的营养代谢物变化与其...     

Ultimate fate of halosulfuron methyl and its effects on enzymatic and microbial activities in three differently textured soils

Halosulfuron methyl is a sulfonylurea herbicide used worldwide for weed control in sugarcane, maize, wheat, and rice production. Considering its environmental impact, this study evaluated the effects of soil type, application rate, and temperature on the dynamics of halosulfuron methyl degradation.Additionally, as soil microbes and enzymes are reliable indicators of the impacts of anthropogenic activities on soil health, the effects of halosulfuron methyl on soil enzymatic and microbial activiti...     

Effects of dietary forage to concentrate ratio and wildrye length on nutrient intake,digestibility, plasma metabolites,ruminal fermentation and fecal microflora of male Chinese Holstein calves

Twenty-eight male, weaned Chinese Holstein calves((156.8±33.4) kg) were used to investigate the effects of dietary forage to concentrate ratio(F:C) and forage length on nutrient digestibility, plasma metabolites, ruminal fermentation, and fecal microflora. Animals were randomly allocated to four treatments in a 2×2 factorial arrangement: whole-length forage(WL) with low F:C(50:50); WL with high F:C(65:35); short-length forage(SL) with high F:C(65:35); and SL with low F:C(50:50). Chinese wildrye was used as the only forage source in this trial. The grass in the SL treatments was chopped using a chaff cutter to achieve small particle size(~50% particles 19 mm). Dry matter intake(DMI) and organic matter(OM) intake was increased by increasing both F:C(P0.01) and forage length(FL)(P0.05), while acid detergent fiber(ADF) and neutral detergent fiber(NDF) intakes were only increased by increasing the F:C(P0.01). The digestibility of NDF was increased as the FL increased(P0.01), and it was also affected by interaction between F:C and FL(P0.05). Cholesterol(CHO)(P0.01), leptin(LP)(P0.05), and growth hormone(GH)(P0.01) concentrations in plasma were increased as dietary F:C increased. A significant increase in plasma triglyceride(TG)(P0.01), insulin(INS)(P0.05), and GH(P0.01) levels was observed with decreasing dietary FL. Ruminal p H values of calves fed with low F:C diets were significantly lower than those in high F:C treatment(P0.05). Increasing the F:C enhanced ruminal acetic acid(P0.05) and acetic acid/propionic acid(P0.01). Fecal Lactobacillus content was significantly higher, while Escherichia coli and Salmonella contents were significantly lower in WL and high F:C groups(P0.05). Lower fecal scores(higher diarrhea rate) were observed in calves fed with SL hay compared to WL hay(P0.05). Denatured gradient gel electrophoresis(DGGE) bands and richness index(S) were significantly affected by the interaction between F:C and FL(P0.05), under high F:C, band numbers and richness index from WL group were higher than that from SL group(P0.05), whereas there were no differences between WL andSL groups under low F:C(P0.05). Microflora similarity was 50–73% among the different treatments. It is concluded that the WL with high F:C(65:35) diet is suitable for weaned calves.     

硅、锌、硼配合氮肥减施对水稻生长的影响

通过田间试验,研究了江汉平原地区氮肥减施条件下,硅、锌、硼等中微量元素配施对水稻生长的影响.结果表明,氮肥在减施20％时,配合硅、锌、硼等中微量元素,不会造成水稻减产,还可以降低穗颈瘟发病率、和优化产量结构.     

初始pH对嗜线虫致病杆菌YL001菌株抑菌活性及代谢的影响

【目的】研究初始培养pH对Xenorhabdus neamatophila YL001发酵液抑菌活性的影响,为进一步提高YL001的抑菌活性提供理论依据。【方法】采用生长速率法、琼脂扩散法、活体组织法,分别测定了初始pH 4.5,5.5,6.5,7.0,7.5和8.5培养条件下,X.neamatophila YL001发酵液及其乙酸乙酯提取物的抑菌活性;同时采用HPLC、LC-MS分析了乙酸乙酯提取物中代谢物的差异。【结果】随着初始pH的升高,共生菌发酵液及其乙酸乙酯提取物的抑菌活性也逐步提高。pH8.5时,YL001发酵液对水稻瘟疫病菌(Magnaporthe grisea)、辣椒疫霉病菌(Phytophthora capsici)、番茄灰霉病菌(Botrytis cinerea)、苹果轮纹病菌(Physalospora piricola)、番茄早疫病菌(Alternaria solani)、甘蓝黑斑病菌(Alternaria brassicae)以及枯草芽孢杆菌(Bacillus subtilis)均具有最强的抑制作用,发酵液乙酸乙酯提取物对番茄灰霉病的EC50为1.29mg/mL,对枯草芽孢杆菌的抑菌圈直径为14.65mm。随着初始培养pH的升高,YL001发酵液及乙酸乙酯提取物对番茄灰霉病的治疗和保护作用也逐渐提高,保护作用明显优于治疗作用。HPLC和LC-MS代谢分析表明,pH8.5、pH 7.0及pH 4.5培养条件下发酵液乙酸乙酯提取物中代谢物含量存在显著差异。【结论】初始培养pH可以显著影响X.neamatophila YL001的抑菌活性及代谢能力,碱性条件更有利于活性物质的产生及其稳定性的保持。