Effects of fixation and storage on flow cytometric analysis of marine bacteria

Flow cytometry (FCM) is now becoming a routine tool for the enumeration and optical characterization of bacteria in marine environments. We investigated the effects of sample fixation and storage upon flow cytometric determination of marine bacteria. Fixed and unfixed seawater samples were analyzed by FCM immediately aboard ship and/or later in the laboratory, and the appearances of the fluorescence signals and bacterial counts of these samples were compared. Fixation and storage led to the formation of multiple peaks in fluorescence histograms; this was also seen in 22 out of 36 samples frozen in liquid nitrogen. Fixation did not, but storage did induce a decrease of bacterial counts: a rapid decrease during the first 3 days followed by a slower decline. The decline of cell numbers in stored samples was expressed by a regression model. Our studies indicate that precaution is necessary when interpreting the data from fixed and/or stored marine bacterial samples analyzed by FCM. The possibility that the procedure of fixation and storage leads to the appearance of high DNA and low DNA bacterial groups should be considered.     

杂色鲍血细胞分类、结构和免疫功能的流式细胞术分析

应用流式细胞术对杂色鲍(Haliotis diversicolor Reeve)血细胞的分类、结构和免疫功能进行分析。根据细胞前向角散射光(FSC)和侧向角散射光(SSC)强度的不同,可将血细胞分为三个亚群:透明细胞、小颗粒细胞和大颗粒细胞,组成比例分别为32.71%、58.17%和8.55%。血细胞的平均总凋亡和死亡率为3.76%。血细胞对荧光微球的总吞噬率为63.67%,其中吞噬1个、2个、3个及以上荧光微球的血细胞分别占22.31%、16.39%、24.96%。线粒体数量、溶酶体数量、非特异性酯酶活性和非诱导性活性氧(ROS)含量均在大颗粒细胞中最高,透明细胞最低。结果表明,杂色鲍三类血细胞在结构和功能上均存在差异,两类颗粒细胞可能在鲍类免疫过程中发挥着更为重要的作用。     

船基围隔条件下沙尘和营养盐添加对近海浮游植物群落结构的影响

为研究沙尘沉降和营养盐输入对中国陆架海域浮游植物群落结构的影响,于2017年3—4月在中国黄、东海进行沙尘和不同营养盐(NO^-₃、PO^-₄、尿素)添加的船基围隔培养实验。结果表明,与对照组相比,沙尘(2 mg/L)和尿素添加实验组的浮游植物群落细胞密度及群落结构变化不显著,叶绿素a含量差异不显著,优势种均为海链藻属 (Thalassiosira)。不同比值的氮、磷无机营养盐添加对水体中叶绿素a含量和细胞密度的影响不同,其中氮磷比为64∶1的实验组叶绿素a含量和细胞密度最高,分别为18.20 μg/L和7.86×10⁵cells/L。沙尘和营养盐添加对浮游植物群落的影响主要表现为叶绿素a含量、细胞密度峰值及不同优势种所占比例的差异,而各个实验组的种类组成及优势种具有一定的相似性。     

基于浮游植物流式细胞仪对胶州湾春季浮游植物的研究

本文基于浮游植物流式细胞仪Cyto Sub对2014年春季胶州湾浮游植物功能群组成及其与环境因子的关系开展研究。Cyto Sub共检测出6个浮游植物类群,分别是聚球藻、微微型真核浮游植物、隐藻、微型单细胞藻、小型单细胞藻和链状藻。多元统计分析显示胶州湾春季浮游植物可划分为两个群落,群落1主要由湾外站位组成,浮游植物生物量浓度为15.15μg/L(以碳含量计,以下同),以小型单细胞藻、链状藻和微型单细胞藻为主;群落2主要由湾内站位组成,浮游植物生物量浓度为72.83μg/L,以链状藻为主。胶州湾的优势类群为链状藻,其丰度与水温和营养盐浓度呈显著正相关关系。与其他浮游植物粒级研究方法的比较表明,当调查海域优势种为小粒径的链状藻时,该方法能够快速、准确地测量出各个浮游植物类群的粒径参数,从而推算出浮游植物群落的粒级结构。     

天然气水合物注热水分解前缘热作用因素分析

为研究天然气水合物注热水分解前缘的热作用影响,建立了考虑孔隙介质中水合物饱和度的分解前缘数学模型,并进行迭代分析求解;实验测温前缘和电阻前缘与数学模型拟合前缘较好的吻合,误差控制在8%以内;分析了注热水温度和速率对分解前缘的热作用影响:温度对分解前缘影响的误差不超过8.71%,较大流速对分解前缘影响误差为4.53%,在提高注入热水温度的同时以较大注入速率能减小对水合物分解前缘的热作用影响。     

Impact of environmental factors on in situ determination of iron in seawater by flow injection analysis

A sensitive method for iron determination in seawater has been adapted on a submersible chemical analyser for in situ measurements. The technique is based on flow injection analysis (FIA) coupled with spectrophotometric detection. When direct injection of seawater was used, the detection limit was 1.6 nM, and the precision 7%, for a triplicate injection of a 4 nM standard. At low iron concentrations, on line preconcentration using a column filled with 8-hydroxyquinoline (8HQ) resin was used. The detection limit was 0.15 nM (time of preconcentration = 240 s), and the precision 6%, for a triplicate determination of a 1 nM standard, allowing the determination of Fe in most of the oceanic regimes, except the most depleted surface waters. The effect of temperature, pressure, salinity, copper, manganese, and iron speciation on the response of the analyser was investigated. The slope of the calibration curves followed a linear relation as a function of pressure (C_p = 2.8 × 10^{− 5}P + 3.4 × 10^{− 2} s nmol^{− 1}, R² = 0.997, for Θ = 13 °C) and an exponential relation as a function of temperature (C_Θ = 0.009e^0.103Θ, R² = 0.832, for P = 3 bar). No statistical difference at 95% confidence level was observed for samples of different salinities (S = 0, 20, 35). Only very high concentration of copper (1000 × [Fe]) produced a detectable interference. The chemical analyser was deployed in the coastal environment of the Bay of Brest to investigate the effect of iron speciation on the response of the analyser. Direct injection was used and seawater samples were acidified on line for 80 s. Dissolved iron (DFe, filtered seawater (0.4 μm), acidified and stored at pH 1.8) corresponded to 29 ± 4% of Fe_a (unfiltered seawater, acidified in line at pH 1.8 for 80 s). Most of Fe_a (71 ± 4%) was probably a fraction of total dissolvable iron (TDFe, unfiltered seawater, acidified and stored at pH 1.8).     

2008年夏季西北冰洋表层水浮游植物群落结构的色素表征及其对不同水团的响应

本文依托2008年夏季中国第三次北极科学考察航次,对西北冰洋海盆区和楚科奇海陆架营养盐及光合色素进行了测定和分析。根据海水理化性质将研究海区分为5个区,并使用CHEMTAX软件（Mackery et al.,1996）讨论了西北冰洋不同海区浮游植物群落组成结构及其与环境因子之间的关系。结果显示在楚科奇海陆架区,太平洋入流显著影响浮游植物生物量和群落结构。高营养盐Anadyr水团以及白令陆架水控制海域,表现出高Chl a且浮游植物以硅藻为主,相反,低营养盐如阿拉斯加沿岸流控制海域,Chl a生物量低且以微型,微微型浮游植物为主。在外陆架海区,海冰覆盖情况影响着水团的物理特征及营养盐浓度水平,相应地显著影响浮游植物群落结构。在海冰覆盖区域,硅藻生物量站到总Chl a生物量的75%以上;在靠近门捷列夫深海平原海区,受相对高盐的冰融水影响（MW-HS）,营养盐浓度和Chl a浓度相对海冰覆盖区略高,浮游植物结构中微型、微微型藻类比重增加,硅藻比例则降至33%;南加拿大海盆无冰海区（IfB）,表层水盐度最淡,营养盐浓度最低,相应地显示出低Chl a生物量,表明海冰消退,开阔大洋持续时间延长,将导致低生物量及激发更小型浮游植物的生长,并不有利于有机碳向深海的有效输出。