螺旋藻藻蓝蛋白对酒精性肝损伤大鼠肠道菌群的影响

酒精性肝病(alcoholic liver disease, ALD)是中国常见的肝脏疾病之一, 严重危害人民健康。目前, 缺乏治疗酒精性肝病的有效方法。螺旋藻藻蓝蛋白(phycocyanin, PC)是一种天然蛋白质, 具有抗炎、抗氧化等作用。本文研究了PC对大鼠酒精性肝损伤的保护作用及其对肠道菌群的影响。利用酒精诱导酒精性肝损伤模型, 将大鼠随机分为4组, 分别为对照组, 模型组, 阳性对照组, 藻蓝蛋白组, 灌胃PC干预之后, 通过肝脏切片的Masson和HE染色的病理分析判断肝损伤的程度, 之后通过16S rRNA高通量测序技术检测大鼠肠道菌群的组成和结构。模型组大鼠肠道菌群发生了紊乱, 显著降低了双歧杆菌(Bifidobacteriaceae)的丰度, 该菌种可以减少肝脏炎症和肝酶, 口服PC后Bifidobacteriaceae的丰度显著增加。此外, 酒精诱导后, 乳酸杆菌(Lactobacillaceae)的丰度略微增加, 促进酒精代谢。藻蓝蛋白能够通过改变酒精性肝损伤大鼠的肠道菌群构成, 从而起到缓解酒精诱导的肝损伤的效果。     

球形赖氨酸芽孢杆菌(Lysinibacillus sphaericus)对斑马鱼幼鱼生长、肠道菌群及代谢的影响

旨在探究养殖水体中添加球形赖氨酸芽孢杆菌对斑马鱼幼鱼生长、抗氧化免疫、肠道菌群与代谢的影响。将初体重为(0.019±0.003) g的270尾斑马鱼随机均分为3组, ZF0组(对照组, 基础饲料)、ZF5组(基础饲料+10⁵CFU/mL球形赖氨酸芽孢杆菌)与ZF7组(基础饲料+10⁷CFU/mL球形赖氨酸芽孢杆菌), 每组3个平行,试验周期为33 d。实验结果表明: (1) 各组间的终末体重、增重率和特定生长率都没有显著性差异(P>0.05)。(2) ZF5和ZF7组的GPT活性显著高于ZF0组(P<0.05), ZF5组AKP的活性显著高于其他组(P<0.05)。(3) ZF7组胰蛋白酶和脂肪酶的活性相对于ZF0和ZF5组显著提高(P<0.05)。(4) ZF7组肠道绒毛高度、V/C都显著高于ZF0与ZF5组(P<0.05); ZF7组的隐窝深度显著低于其他两组(P<0.05)。(5) 从肠道菌群组成来看, 各组间优势菌门均为变形菌门(Proteobacteria), ZF5和ZF7组显著降低了肠道中梭杆菌门(Fusobacteriota)的丰度并显著提高了放线菌门(Actinobacteriota)的丰度(P<0.05)。属水平上, 对照组与试验组的优势菌属为鲸杆菌属(Cetobacterium), 试验组显著减少了邻单胞菌属(Plesiomonas)的丰度同时显著提高了戈登氏菌属(Gordonia)、细杆菌属(Microbacterium)等有益菌属的丰度(P<0.05)。(6)ZF5和ZF7组与ZF0组的肠道代谢物差异明显, 其中Lipoxin a4、Prostaglandin g2和11-dehydrothromboxane b2显著上调(P<0.05), 5''-phosphoribosyl- 5-amino-4-imidazolecarboxamide (aicar)显著下调(P<0.05)。差异代谢物主要富集在ABC转运蛋白、神经活性配体-受体相互作用以及丙氨酸、天冬氨酸和谷氨酸通路中。综上所述,养殖水体中添加球形赖氨酸芽孢杆菌有利于改善斑马鱼幼鱼的肠道消化性能、组织结构和微生物多样性及其代谢产物。     

3月龄脉红螺表型性状对体质量影响效应分析

随机抽取3月龄脉红螺稚螺103粒,开展其表型性状对体质量性状影响分析,以期为脉红螺人工选择育种提供较好的测度指标。实验测量了脉红螺壳长（X₁）、壳宽（X₂）、壳厚（X₃）、壳口长（X₄）、壳口宽（X₅）等5种表型性状和体质量性状（Y）,运用相关性分析、多元回归分析、通径分析及决定系数分析等方法分析了表型性状对体质量性状的影响。结果表明,脉红螺各性状间的相关性均达到极显著水平（P<0.01）,拟合的多元线性方程为Y=-18.484 4+0.348 35X₁+0.463 79X₂,经自助法检验,在95%置信区间条件下,该方程决定系数（R²）范围为0.863 5~0.939 4,拟合度较好;壳长和壳宽对体质量的决定系数之和大于0.85,是最适宜用来预测体质量性状的两个表型性状指标。本研究成功分析出对脉红螺体质量直接影响作用最强的表型性状,为脉红螺选择育种理想测度指标的挑选提供了依据。     

苦参汤对皮肤溃疡大鼠模型创口愈合的影响

目的：研究苦参汤对皮肤溃疡大鼠伤口愈合的影响。方法：通过皮下注射75%冰醋酸诱导大鼠皮肤溃疡模型,将模型大鼠分为5组,G1采用蒸馏水干预,G2～5分别采用苦参汤低、中、高剂量及0.02%高锰酸钾溶液（PP）进行局部治疗,连续7d。治疗后对各组溃疡面积、溃疡评分和组织学评估,通过免疫组织化学分析检测前炎性细胞因子前列腺素E₂（PGE-2）和白细胞介素-8（IL-8）的表达。结果：苦参汤对皮肤溃疡大鼠创口愈合有促进作用,中剂量和高剂量苦参汤治疗的大鼠溃疡愈合更佳（P<0.05）,低剂量苦参汤治疗的肉眼可见溃疡评分显著降低（P<0.01）,PP组在降低溃疡面积和肉眼可见的溃疡评分方面,差异无统计学意义（P>0.05）;所有剂量的苦参汤和PP均改善了大鼠诱导的皮肤溃疡的组织学损伤;高剂量苦参汤和PP使大鼠皮肤溃疡组织中的PGE-2和IL-8表达显著降低（P<0.01）;各组大鼠均未发现与治疗有关的毒性。结论：苦参汤可促进皮肤溃疡大鼠的伤口愈合,其对PGE-2和IL-8的抑制是增强伤口愈合的可能作用机制。     

中国黄渤海真江蓠(Agarophyton vermiculophyllum)群体遗传多样性研究

真江蓠是原产于西北太平洋的重要经济红藻。我们利用10对微卫星引物检测中国黄渤海地区真江蓠的群体遗传多样性和结构。10个微卫星位点在12个群体中共检测到65个等位基因,每个位点的等位基因（N_a）为1~28,有效等位基因（N_e）为1.0~9.6。每个群体的平均等位基因（N_a）、平均有效等位基因（N_e）、平均香浓指数（I）、平均观察杂合度（H_o）和平均预期杂合度（H_e）分别为2.4、1.6、0.419、0.133和0.227,显示较低的群体遗传多样性。中国黄渤海12个真江蓠群体间遗传分化较大（F_st=0.398 7）,基因流有限（N_m=0.377 1）,近交系数为正（F_is=0.391 3,F_it=0.634 0）,表明可能存在近交和杂合子缺失现象。Structure和UPGMA系统进化分析一致将12个群体分为两个遗传组,并在黑石礁群体（HS）和石岛群体（SD）中发现明显的遗传混杂现象。AMOVA分析显示遗传变异主要来自于群体内（73.27%）。该研究可为黄渤海地区真江蓠自然资源保护和管理提供科学依据。     

珠江口溶解有机物的遥感动态监测

河口有色溶解有机物（colored dissolved organic matter,CDOM）的分布是各种物理-生物地球化学过程共同作用的结果。为实现河口高动态变化CDOM的监测,遥感是一种重要的手段。由珠江口四个不同季节的航次获得的实测数据,本文构建了一个遥感算法以反演CDOM在400 nm的吸收系数（a_CDOM （400））。该算法使用以波段反射率比值R_rs （667）/R_rs （443）和R_rs （748）/R_rs （412）为自变量。将构建的算法应用于2002-2014年的MODIS/Aqua数据,本文计算了珠江口不同季节的a_CDOM （400）气候态分布。CDOM的分布主要受珠江径流量和区域水下地形特征的影响。沿着垂直于水深梯度的断面,气候态a_CDOM （400）呈指数减少（y=ae^bx,b<0）,但不同季节差异很大。珠江口CDOM主要是河流淡水输运而来。其中,富里酸比例随盐度的增加而降低。基于构建的算法、CDOM保守混合方程和径流量,本文由MODIS/Aqua数据进一步估算了2002-2014年夏季和冬季珠江DOC的有效入海浓度和有效入海通量。珠江的有效入海浓度和有效入海通量都与流量存在正相关关系,且在夏季的相关性更明显,R2分别为0.698和0.9657。     

牙鲆gnrh基因的表达分析

本文通过qPCR方法,研究了我国重要海水鱼类——牙鲆（Paralichthys olivaceus）雌、雄成体组织以及性腺分化期的脑、性腺中gnrh1、gnrh2和gnrh3的表达水平。结果显示,gnrh1分布于所有检测的组织：在脑、垂体、肾脏和肝脏中高表达,在性腺中表达相对较低;gnrh2则在雌、雄性的肝脏以及雄性的肠、肾脏、眼睛和头肾中;而gnrh3只在雄性的肾脏、眼睛和肌肉中检测到微弱表达。人工诱导的牙鲆雌核发育鱼苗分别用17β-雌二醇（E₂）和17α-甲基睾酮（MT）处理,使其分化为雌性或雄性个体表型。在性腺分化期的脑中,gnrh1的表达呈现上升趋势,在全长（total length,TL）4 cm鱼苗中,E₂组的表达显著高于MT组（P<0.05）;gnrh2在2 cm TL时E₂组显著高于MT组（P<0.05）,随后在E₂和对照组的表达水平均出现下降趋势;gnrh3在2 cm TL时E₂和对照组的表达均显著高于MT组（P<0.05）,自6 cm TL时MT和E₂组的表达开始下降。在性腺中,三种gnrh在性腺分化前2 cm TL表达量都相对较高,随后呈现下降趋势。综上,推测牙鲆gnrh1可能参与了性腺分化的启动、分化过程及性腺发育,gnrh2主要参与了性腺分化的启动,gnrh3主要参与了性腺分化的启动和分化过程。     

褐菖鲉(Sebastiscus marmoratus)幼鱼生物学表型对体质量的影响效应

探究影响水产养殖动物体质量的关键生物学表型性状组合,进而揭示其体质量增长对策,对指导水产养殖动物的种质甄别与科学养殖具重要现实意义。随机选取象山西沪港海域板式网箱内养殖3个月的褐菖鲉幼鱼同生群个体60尾为研究对象,以全长(X₁)、体长(X₂)、体宽(X₃)、体高(X₄)、头宽(X₅)、侧线长(X₆)、尾柄高(X₇)、体质量(BW)、净体质量(NW)、内脏质量(W₁)、肠质量(W₂)、肝质量(W₃)、胃质量(W₄)、鳃质量(W₅)、心质量(W₆)为生物学测定指标,采用多元分析方法定量研究了体尺性状和称量性状对体质量的影响效应。结果表明:(1)经统计,所涉称量性状测定值较体尺性状更显离散,两者变异系数的波动范围分别为38.27%~89.86%和13.86%~17.78%,排序分别为W₄>W₁>W₆>W₂>W₅>W₃>NW>BW和X₃>X₄>X₆>X₅>X₂>X₇>X₁;经计算,消化脏器与内脏间的质量比呈W₃>W₄>W₂ (P<0.05);(2)相关分析显示,所涉体尺性状和称量性状均与BW呈极显著相关(P<0.01),两者相关系数波动范围分别为0.414~0.990和0.899~0.961,排序分别为NW>W₃>W₁>W₆>W₅>W₂>W₄和X₁>X₄>X₂>X₅>X₆>X₃>X₇;(3)经通径分析,被保留的称量性状组合和体尺性状组合的直接作用排序分别为NW>W₃>W₂>W₄>W₆和X₄>X₅>X₆>X₇;经决定程度分析和复相关分析,称量性状组合对BW的总决定系数和复相关指数均为0.989,体尺性状组合则均为0.961;(4)经偏回归分析,分别获得了基于称量性状组合和体尺性状组合的BW估算方程。研究结果可为褐菖鲉速生型种质遴选与科学养殖提供参考。     

黄条（鱼师）(Seriola aureovittata)生长对工厂化养殖密度的微生态适应特性

研究低(5 kg/m³)、中(10 kg/m³)、高(15 kg/m³)三个养殖密度对工厂化养殖黄条（鱼师）生长的影响, 从消化道菌群角度分析了养殖黄条（鱼师）的生长的密度适应性。利用Illumina MiSeq平台对不同养殖密度条件下黄条（鱼师）的消化道(胃、肠道、幽门盲囊)菌群进行16S rRNA高通量测序, 并进行了菌群的多样性、结构特征和菌群基因功能预测的分析。结果显示, 相较于中、低密度组, 高养殖密度显著(P<0.05)抑制了黄条（鱼师）的终末体质量、特定生长率并显著上调了血清应激相关激素水平。消化道微生物群落在属水平上, 乳杆菌属(Lactobacillus)、鞘氨醇单胞菌属(Sphingomonas)、双歧杆菌属(Bifidobacterium)、Faecalibaculum、Ruminococcaceae_UCG等在中、高密度组中丰度降低, 克雷伯氏菌属(Klebsiella)等在中、高密度组中丰度增加, 均与低密度组差异显著(P<0.05)。KEGG通路分析发现, 各养殖密度组的菌群基因主要集中在代谢、环境信息过程、遗传信息过程三个一级功能通路。该实验设置的中、高密度组相对低密度组, 消化道菌群的组成和结构上呈现有益菌(乳酸杆菌属等)显著性减少和条件致病菌(克雷伯氏菌属)显著性增加(P<0.05)的趋势。在菌群功能方面, 与低密度组相比, 中、高密度组消化道特别是在代谢、遗传及环境信息过程的二级功能通路上注释的tags数目相对低密度组显著增加(P<0.05)。研究结果为建立工厂化养殖模式下适宜密度调控和健康养殖技术建立提供了理论依据。     

藻蓝蛋白对博来霉素肺纤维化小鼠肠道菌群的影响

博来霉素(bleomycin,BLM)是目前用于治疗头颈癌等多种癌症的药物之一,但会产生一系列副作用,例如肺纤维化(pulmonary fibrosis,PF),这大大降低了患者的生活质量。藻蓝蛋白(phycocyanin,PC)作为一种色素蛋白对包括PF在内的多种疾病具有调节作用。研究发现,慢性肠道疾病通常伴有肺部疾病,患者的肠道菌群会发生紊乱。针对上述线索,研究了PC对BLM诱导的PF小鼠肠道菌群的影响。通过尾静脉将BLM注入小鼠体内,以构建肺纤维化模型。然后通过16S r RNA高通量基因测序技术,分析了PC对BLM引起的PF小鼠肠道菌群的影响。结果表明,PC降低了BLM诱导的PF程度并调节了小鼠的肠道菌群。通过尾静脉注射BLM后,小鼠肺部受到损伤,肠道菌群发生紊乱。口服藻蓝蛋白干预后,小鼠的肺组织切片显示其胶原蛋白沉积减少,肺泡间隔和炎性细胞浸润减少,小鼠PF程度减轻。此时,小鼠肠道菌群中Alistipes、Lachnospiraceae等有益菌的丰度增加,Rikenella等有害菌的丰度降低,肠道菌群趋于正常。推测PC可以通过增加肠道中有益细菌的数量并减少肠道中有害细菌的数量...     

Microbial Biomass and Organic Nutrients in the Deep-Sea Sediments of the Central Indian Ocean Basin

In order to assess the impact of deep-sea mining on the in situ benthic life, we measured the microbial standing stock and concentration of organic nutrients in the deep-sea sediments of the Central Indian Ocean Basin in the Indian pioneer area. Sediments were collected using box core and grab samples during September 1996. The total bacterial numbers ranged from 10 10 -10 11 cells per g -1 dry weight sediment. There was a marginal decrease in the number of bacteria from surface to 30 cm depth, though the subsurface section registered a higher number than did the surface. The highest numbers were encountered at depths of 4-8 cm. The retrievable number of bacteria were two orders less in comparison with the direct total counts of bacteria. An almost homogeneous distribution of bacteria, total organic carbon, living biomass, and lipids throughout the depth of cores indicates active microbial and benthic processes in the deep sea sediments. On the other hand, a uniform distribution of total counts of bacteria, carbohydrates, and total organic carbon in all the cores indicates their stable nature and suggests that they can serve as useful parameters for long-term monitoring of the area after the benthic disturbance. Further studies on temporal variability in this region would not only verify the observed norms of distribution of these variables but would also help to understand restabilization processes after the simulated benthic disturbance.     

Assessing the Validity of Seismo-Acoustic Predictor Equations for Obtaining Seabed and Sub-Surface Sediment Physical Properties

The interdependence between the seismo-acoustic properties of a marine sediment and its geotechnical/physical parameters has been known for many years, and it has been postulated that this should allow the extraction of geotechnical information from seismic data. Though in the literature many correlations have been published for the surficial layer, there is a lack of information for greater sediment depths. In this article, a desktop study on a synthetic seafloor model illustrates how the application of published near-surface prediction equations to subsurface sediments (up to several tens of meters burial depth) can lead to spurious predictions. To test this further, acoustic and geotechnical properties were measured on a number of sediment core samples, some of which were subjected to loading in acoustically-equipped consolidation cells (oedometers) to simulate greater burial depth conditions. For low effective pressures (representing small burial depths extending to around 10 meters subsurface), the general applicability of established relationships was confirmed: the prediction of porosity, bulk density, and mean grain size from acoustic velocity and impedance appears generally possible for the investigated sedimentary environments. As effective pressure increases through, the observed relationships deviate more and more from the established ones for the near-surface area. For the samples tested in this study, in some instances increasing pressure even resulted in decreasing velocities. There are several possible explanations for this abnormal behavior, including the presence of gas, overconsolidation, or bimodal grain size distribution. The results indicate that an appropriate depth correction must be introduced into the published prediction equations in order to obtain reliable estimates of physical sediment properties for greater subsurface depths.     

A Review of Some Environmental Issues Affecting Marine Mining

This article reviews information recently available from existing marine and coastal mining for responses to environmental issues affecting marine mining at different depths. It is particularly but not exclusively concerned with those issues affecting seabed biodiversity impact and recovery. Much information has been gathered in the past 10 years from shallow mining operations for construction aggregate, diamonds, and gold, from coastal mines discharging tailings to shallow and deep water, and from experimental deep mining tests. The responses to issues identified are summarized in a series of eight tables intended to facilitate site-specific consideration. Since impacts can spread widely in the surface mixing layer SML, and can affect the biologically productive euphotic zone, the main issues considered arise from the depth of mining relative to the SML of the sea. Where mining is below the SML, the issue is whether it is environmentally better to bring the extraction products to the surface vessel for processing (and waste discharge), or to process the extraction products as much as possible on the seabed. Responses to the issues need to be sitespecific, and dependent on adequate preoperational environmental impact and recovery prediction. For deep tailings disposal from a surface vessel, there are four important environmental unknowns: (1) the possible growth of "marine snow" (bacterial flocs) utilizing the enormous quantities of fine tailings particles (hundreds or thousands of metric tons per day) as nuclei for growth, (2) the possibility that local keystone plankton and nekton species may migrate diurnally down to and beyond the depth of deep discharge and hence be subjected to tailings impact at depth, (3) the burrow-up capability of deep benthos and their ability to survive high rates of tailings deposition, and (4) the pattern and rate of dispersion of a tailings density current through the deep water column from discharge point to seabed. Actions to obtain relevant information in general and site-specifically are suggested.     

Increased Particle Fluxes at the INDEX Site Attributable to Simulated Benthic Disturbance

Particle fluxes were measured 7 m above the sea bottom during the predisturbance, disturbance, and postdisturbance periods by using time series sediment traps attached to seven deep-sea moorings deployed in the INDEX experiment site in the Central Indian Basin. The predisturbance particle fluxes varied between 22.3 to 55.1 mg m -2 day -1 . Increased and variable particle fluxes were recorded by the sediment traps during the disturbance period. The increase observed was 0.5 to 4 times more than the background predisturbance fluxes. The increases in particle fluxes (~4 times) recorded by the sediment trap located in the southwestern direction (DMS-1) were the greatest, which could be the result of preferential movement of resuspended particles generated during the deep-sea benthic disturbance along the general current direction prevailing in this area during the experimental period. Also, the traps located closer to the disturbance area recorded greater fluxes than did the traps far away, across the Deep Sea Sediment Resuspension System path. This variability in recorded particle fluxes by the traps around the disturbance area clearly indicates that physical characteristics such as grain size and density of the resuspended particles produced during the disturbance had an important effect on particle movement. The postdisturbance measurements during ~5 days showed a reduction in particle fluxes of ~50%, indicating rapid particle settlement.     

Geoacoustic Inversion via Genetic Algorithm and Its Application to Manganese Sediment Identification

An acoustic inversion method using a wide-band signal and two near field receivers is proposed and applied to multiple layered seabed models including a manganese sediment. The inversion problem can be formulated into a probabilistic model comprised of signals, a forward model, and additive noise. The forward model simulates wide-band signals, such as chirp signals, and is chosen to be the source-waveletconvolution plane wave modeling method. The wavelet matching technique, using weighted least-squares fitting, estimates the sediment sound-speed and thickness on which determination of the possible numerical ranges for a priori uniform distribution is based. The genetic algorithm is applied to a global optimization problem to find a maximum a posteriori solution for determined a priori search space. Here the object function is defined by an L 2 norm of the difference between measured and modeled signals. Not only the marginal pdf but also its statistics are calculated by numerical evaluation of integrals using the samples selected during importance sampling process of the genetic algorithm.     

海上大直径钢管桩打桩过程中桩周土强度弱化研究

海上大直径钢管桩打桩过程中,桩周土体受到强烈扰动而发生强度弱化,掌握桩周土体强度弱化规律对于准确预测打桩过程、保证工程安全具有重要意义。为研究土体强度弱化规律,开展了环剪试验模拟打桩对桩周土体的扰动,测试土体强度随剪切速率的变化规律,建立了描述土体强度弱化规律的拟合公式,引入到打桩分析软件中。研究结果表明：土体的强度折减程度不仅与土体本身的性质有关还受到土体的埋深和剪切速率的影响,埋深越深土体强度折减程度越低,剪切速率越高土体强度折减越高,在打桩分析中可采用这里推荐的线性折减方法来模拟不同深度处土体强度的折减规律。     

Determination of In Situ Sediment Shear Strength from Advanced Piston Corer Pullout Forces

The advanced piston corer (APC) has been used by the Ocean Drilling Program since 1985 for recovering soft sediments from the ocean floor. The pullout force measured on extracting the core barrel from the sediment is shown to correlate with the average shear strength of the sediment core measured in the ship's laboratory. A simple rule of thumb is derived relating the shear strength of the sediment to the pullout force. Multiple APC holes at individual sites allow the consistency of the pullout measurements to be assessed. The effects of different operational procedures during APC coring are also explored. Although generally applicable, the correlation between pullout force and laboratory measurements of shear strength breaks down for some APC holes, possibly because of the disturbance of some sediment types during the APC coring process. A better understanding of the physical process of APC coring, and its effect on the properties of the sediment both inside and immediately outside the core barrel, would indicate what confidence can be put on the measurement of pullout force as a way of evaluating the in situ shear strength of deep sea sediments.     

Interannual Sea Level Variations and Annual Tides in the Northwestern Pacific

Specific properties of the interannual sea level variations and annual tides in the Northwestern Pacific were studied. Several tide stations were monitored. The monthly mean sea level for the year of 1995 was analyzed at each tide station. A seismic event in 1995, some tectonic activity around the subject area, and the Kuroshio (the oceanic western boundary current) may possibly contaminate results which would have occurred from the astronomical annual tide alone.     

Flow Cytometric Analysis of Coastal Lagoon Bacterioplankton and Picophytoplankton: Fixation and Storage Effects

The effect of different fixation and storage protocols on the flow cytometric (FCM) simultaneous analysis of bacterioplankton and phytoplankton in coastal seawater samples (Mediterranean coastal lagoons) was investigated. FCM measurements (cell number, fluorescence and scatter characteristics) were obtained through DAPI staining. Three fixatives [glutaraldehyde (GA), formaldehyde (FA) and paraformaldehyde (PFA)] and two storage (3 months duration) methods (5 °C and −196 °C) were tested. Two dominant populations were detected in studied samples: bacteria and eukaryotic picophytoplankton. Adding fixatives (2% final concentration) appears necessary to obtain FCM exhaustive counts of all the bacteria and phytoplanktonic cells. This was related to the permeation effect of fixatives which allowed a better DAPI staining of the cells. Maximum fluorescence, i.e. optimal staining of the cells was obtained with FA or PFA, and significant lower fluorescences with GA. Fixed samples stored at 5 °C induced rapid cell loss. Only storage in liquid nitrogen of samples fixed with FA or PFA, allows mid-term (≥4 months) preservation of bacteria or picophytoplankton cell numbers, and limited evolution of DAPI-induced fluorescence and scatter characteristics.