Continuous live cell imaging of cellulose attachment by microbes under anaerobic and thermophilic conditions using confocal microscopy

Live cell imaging methods provide important insights into the dynamics of cellular processes that cannot be derived easily from population-averaged datasets.In the bioenergy field,much research is focused on fermentation of cellulosic biomass by thermophilic microbes to produce biofuels;however,little effort is dedicated to the development of imaging tools to monitor this dynamic biological process.This is,in part,due to the experimental challenges of imaging cells under both anaerobic and thermophilic conditions.Here an imaging system is described that integrates confocal microscopy,a flow cell device,and a lipophilic dye to visualize cells.Solutions to technical obstacles regarding suitable fluorescent markers,photodamage during imaging,and maintenance of environmental conditions during imaging are presented.This system was utilized to observe cellulose colonization by Clostridium thermocellum under anaerobic conditions at 60℃.This method enables live cell imaging of bacterial growth under anaerobic and thermophilic conditions and should be widely applicable to visualizing different cell types or processes in real time.     

厦门市郊区降尘中大分子有机质化学特征浅析

以厦门市的降尘(包括一次沙尘天气)为研究对象,采用综合湿法-化学方法获得腐殖酸(HA)、干酪根+碳黑(KB)和碳黑(BC)3类大分子有机质,分析了两种天气情况下大分子有机质的形貌特征、官能团结构和来源,同时探讨沙尘天气条件下的影响.结果表明,大分子有机质的形貌特征为探讨其来源提供了丰富的信息,如呈大孔网状结构的干酪根样品可能源于植物,而呈球状结构的BC样品则可能源于煤或油的不完全燃烧.傅立叶红外光谱研究表明,降尘中的HA、KB和BC具有相似的结构组成和官能团信息,而受到沙尘天气及季节性的影响,官能团的含量存在差别.碳稳定同位素结合扫描电镜进一步研究大分子有机质来源表明,大分子有机质有明显的陆源C3植物的信息,且有部分化石燃料的信息.沙尘天气降尘中大分子有机质的δ13C值低于正常天气,这与植物的不同地域生长环境、沙尘暴携带的来自蒙古沙漠和中国西北部的沙尘及陆源有机质的δ13C值偏低区域的沙尘影响有关.KB和BC的相关性显著(r=0.964,p ＜0.01),表明干酪根(K)与BC有部分相似的生物质来源.     

Cantilever nanobiosensor using tyrosinase to detect atrazine in liquid medium

The aim of this study was to develop a cantilever nanobiosensor for atrazine detection in liquid medium by immobilising the biological recognition element (tyrosinase vegetal extract) on its surface with self-assembled monolayers using gold, 16-mercaptohexadecanoic acid, 1-ethyl-3-[3-dimethylaminopropyl] carbodiimide hydrochloride/n-hydroxysuccinimide. Cantilever nanobiosensors presented a surface compression tension increase when atrazine concentrations were increased, with a limit of detection and limit of quantification of 7.754 ppb (parts per billion) and 22.792 ppb, respectively. From the voltage results obtained, the evaluation of atrazine contamination in river and drinking water were very close to those of the reference sample and ultrapure water, demonstrating the ability of the cantilever nanobiosensor to distinguish different water samples and different concentrations of atrazine. Cantilever nanosensor surface functionalization was characterised by combining polarisation modulation infrared reflection-absorption spectroscopy and atomic force microscopy and indicating film thickness in nanometric scale (80.2 ± 0.4 nm). Thus, the cantilever nanobiosensor developed for this study using low cost tyrosinase vegetal extract was adequate for atrazine detection, a potential tool in the environmental field.     

常温ANAMMOX工艺运行性能及功能菌研究

通过改变进水基质浓度,对以火山岩为填料的常温上向流厌氧氨氧化生物滤池在不同基质浓度下的脱氮性能进行了研究.借助显微镜、电镜(SEM)观察对滤池内的微生物形态进行了分析,利用变性凝胶电泳(DGGE)和克隆测序等微生物方法,对ANAMMOX种属进行了鉴定.试验结果表明,常温条件下,厌氧氨氧化滤池在高、低不同基质下都能够保持较高的脱氮效果,其中TN去除负荷最高能够达到2.99kgN/(m3×d),NH4+-N和NO2--N最高去除率分别能够达到99.4%和98.8%;显微镜、电镜观察显示:滤池下部的微生物种类更丰富,ANAMMOX菌在滤池的中部和上部的比例更高;16SrRNA克隆测序鉴定结果表明:滤池中的ANAMMOX菌为Candidatus Kuenenia stuttgartiensis,其对温度和基质浓度都有着较宽的适应性.     

Novel method for in situ visualization of polycyclic aromatic hydrocarbons in mangrove plants

Two-photon laser confocal scanning microscopy (TPLCSM) was first used to visualize the uptake and movement of naphthalene (NAP), phenanthrene (PHE), and pyrene (PYR), from separately contaminated hydroponics solutions into living Kandelia candel (L.) Druce seedlings. With this non-destructive and non-chemical extraction technique, the experimental results revealed that three types of polycyclic aromatic hydrocarbons (PAH) were observed moving into the K. candel and its translocation to radicle, hypocotyl and leaf, and the transmission rates of these PAH in the K. candel were in the order of NAP?>?PHE?>?PYR. Data demonstrate the manner in which PAH enter, transport, and distribute within the K. candel, and provided us some valuable information on uptake and translocation mechanism of PAH. These findings may help to optimize the phytoremediation strategies of PAH in mangrove wetlands.     

Noise exposure of pregnant mice induces heart defects in their fetuses

To determine the effects of noise exposure on hearts of newborn mice, 48 pregnant mice were exposed to recorded aircraft flight noise (100?dB) for 150?min from the 2nd day of pregnancy to 7th day after delivery. Mice of the same age were selected as control group. The weights and crown-rump lengths of the fetuses and the newborns were measured, and their hearts were removed under deep anesthesia on the 1st, 7th, and 14th days after birth and prepared histologically. Quantitative changes of the hearts were measured stereologically by the Cavalieri method. The ultrastructural changes of cardiac myocytes were examined using transmission electron microscopy. No histological changes were observed, but the volumes of newborns’ hearts in the experimental groups increased significantly compared to the control group. Photomicrographs of cardiac muscle cells revealed nuclear envelope changes, chromatin condensation, and increased mitochondria and myofibrils in noise-exposed newborns. It is concluded that the newborns of pregnant mice exposed to noise stress have hypertrophic hearts with ultrastructural changes.     

Effect of Cd++ on germination,growth and active principles of Datura metel L.∗

The effects of Cd⁺⁺ concentration increment in soil and/or in atmosphere, on germination, growth and alkaloid contents of Datura metel L. were investigated. We observed a significant reduction of growth following addition of Cd⁺⁺ in soil. Minor effects on growth were evident when the plants were sprayed with CdCl₂ solution (10mgCd⁺⁺/l).

Scanning electron microscope observation showed stomata widely open and conspicuous cuticular striations in the foliar epidermis of polluted plants.

GLC analysis of extracts evidenced a significant variation in active principle content among plants undergoing different treatments.     

硫酸根离子与聚合铝的反应过程及其产物形态

对不同Al/SO2-4比及碱化度条件下PACl与SO₄^2-的反应过程进行研究,分析了体系中总铝浓度及形态分布变化规律,测定了各沉淀/结晶产物的组成,并对其形貌进行了扫描电镜观察.实验结果表明:碱化度对PACl/SO₄^2-反应体系有较大影响,随着碱化度(B值)的增加,沉淀/结晶的析出速率加快,结果析出物中OH含量逐渐增加,而SO₄^2-/Al比率从0.45～0.30逐渐降低.各聚合铝水解形态与SO₄^2-反应速率上存在明显差异,从而为其混凝优势形态的分离纯化创造了条件.     

Changes of Surface Properties of Yeast Cell Wall Under Exposure of Electromagnetic Field (40.68 MHz) and Action of Nystatin

Summary Changes of the surface properties were studied in the cell wall of the yeast Saccharomyces cerevisiae Y-517 under influence of the electromagnetic field (EMF) (40.68 MHz) and lethal doses of the fungicidal antibiotic nystatin (10 μg/10⁶ cells). Atomic force microscopy was used to study surface topography and visco-elastic properties of the cell walls. Surface carbohydrates were detected by lectins marked with gold with the help of the scanning electron microscope. Additional polysaccharide layer appeared over cell surface after EMF exposure. We suggest that electromagnetic fields resulted in the change of the cell surface, and, accordingly, the sensitivity of organisms to the antifungal antibiotics.