城市污水厂尾水对文蛤微核率、溶酶体膜稳定性及解毒酶活性的影响

利用天然海水将取自青岛市团岛污水处理厂的尾水稀释为不同浓度(EVR=1%、5%、10%、20%、30%、40%)的混合液,对文蛤(Meretrix meretrix)进行15d的暴露培养,并测定血细胞中的溶酶体膜稳定性(LMS)、微核率(MNF)以及鳃和内脏中I相解毒酶(7-乙氧基-3-异吩恶唑酮-脱乙基酶-EROD)和Ⅱ相解毒酶(谷胱甘肽转硫酶-GST)随EVR的变化。结果表明,各尾水处理组(即使EVR=1%)的血细胞LMS均明显低于对照水平(P0.05);EVR≥20%的尾水造成血细胞MNF的显著诱导(P0.05);然而,鳃、内脏中的EROD、GST活性仅在少数尾水处理组中被激活(P0.05)。文蛤血细胞LMS对尾水暴露的响应敏感性及其与EVR之间的显著负相关性(R=-0.857,P0.05),使其适于作为纳污海域尾水污染的生物标志物。     

褐牙鲆卵黄原蛋白免疫胶体金技术的开发及其在环境雌激素检测中的应用

为快速检测海洋环境雌激素,本研究利用20 nm胶体金颗粒和褐牙鲆(Paralichthys olivaceus) Vtg单/多克隆抗体开发了夹心法免疫胶体金检测试纸条,并对胶体金标记条件、T线和C线抗体浓度等参数进行了优化,构建了褐牙鲆Vtg的快速检测技术。实验结果表明该胶体金试纸条工作范围为15.7～1 000 ng·mL^-1(R²>0.98),检出限为7.8 ng·mL^-1。本研究制备的胶体金试纸条检出限与目前常用的多种Vtg的ELISA方法相近,但是检测时间仅为15 min,比常规方法节省了约14 h。此外,该试纸条在存放30和90 d后,对Vtg的检测结果并无明显偏差,表现出较好的稳定性和可重复性。最后,通过测定多种常见环境雌激素(双酚A、雌二醇、乙炔雌二醇)暴露对雄性成年褐牙鲆血浆中Vtg的诱导情况,进一步验证了该试纸条在海洋环境雌激素活性检测中的应用。     

金鱼卵黄原蛋白单抗夹心ELISA的开发及其在环境雌激素检测中的应用

利用Western blot和ELISA方法,从实验室现有的斑马鱼(Danio rerio)卵黄脂磷蛋白(Lipovitellin,Lv)单克隆抗体库中筛选出了对金鱼(Carassius auratus)卵黄原蛋白(Vitellogenin,Vtg)具有高特异性、高亲和力的单抗H3A8。以纯化的单抗H3A8、金鱼Lv与HRP标记的Lv多克隆抗体建立了夹心ELISA,其工作范围为15.6~1 000ng·mL-1,检出限为9.6ng·mL-1,组内差异与组间差异分别为1.30%~4.99%和2.30%~4.37%,并且金鱼Vtg与Lv的标准曲线几乎重合,表明建立的夹心ELISA可以准确定量金鱼Vtg,并利用建立的ELISA测定了10、100、1 000ng/L 17β-雌二醇(17β-estradiol,E2)暴露3和21d后雄性金鱼血浆与体表粘液中的Vtg含量,发现1 000ng·L-1 E2暴露3d、100与1 000ng·L-1 E2暴露21d均能显著升高雄鱼血浆和体表粘液中的Vtg含量,E2暴露21d后雄鱼血浆与体表粘液中的Vtg含量较为接近,建议将取样简便且对鱼体无伤害的体表粘液用作今后金鱼Vtg的检测样品。     

大泷六线鱼卵黄原蛋白的纯化鉴定及其ELISA检测方法的建立

海洋环境雌激素污染日益加剧,亟需建立可靠的生物检测技术,鱼类卵黄原蛋白(vitellogenin, Vtg)是国际上推荐的环境雌激素生物标志物。本研究采用凝胶过滤与离子交换层析技术从17β-雌二醇(E_2)肌肉注射后的雄性大泷六线鱼(Hexagrammos otakii)血浆中纯化获得Vtg。经鉴定大泷六线鱼Vtg是分子量约为340 kDa的糖磷脂蛋白,SDS-PAGE显示其亚基分子量约为170 kDa。利用纯化的Vtg免疫新西兰大白兔制备了多克隆抗血清,并且Western blot结果证实抗血清对Vtg具有很好的特异性。以纯化的Vtg与兔抗Vtg多克隆抗体建立了检测大泷六线鱼血浆Vtg的间接竞争酶联免疫吸附反应(ELISA)。该检测方法的工作范围为15.62～1 000 ng/mL,组内与组间差异分别为6.85%与6.79%,可用于大泷六线鱼血浆Vtg的准确定量。本研究建立的大泷六线鱼Vtg ELISA具有较高的敏感度、特异性与精确度,为中国海洋环境雌激素的生物监测提供了可靠工具。     

酚类污染物对金鱼卵黄蛋白原诱导的雌激素效应研究

以雌二醇为阳性对照,通过雄性金鱼卵黄蛋白原的诱导实验研究壬基酚、辛基酚、双酚A和2,4-二氯酚4种酚类污染物的雌激素活性,同时探讨金鱼作为实验生物的敏感性。不同浓度梯度的酚类污染物暴露14d后,以酶联免疫吸附法(ELISA)测定金鱼血浆中的卵黄蛋白原含量,同时测定肝脏指数。结果表明,4种酚类污染物均可诱导金鱼体内卵黄蛋白原的合成并且提高其肝脏指数。雌二醇、壬基酚、辛基酚和双酚A诱导雄性金鱼卵黄蛋白原合成的最低可见效应浓度分别为0.005,15,5,10μg/L,且诱导量随暴露浓度的升高呈明显的剂量-效应关系,可以采用Weibull函数进行非线性拟合,其半效应浓度EC50分别为0.078,255.7,79.6和113.4μg/L。2,4-二氯酚诱导能力较弱,最大诱导量比阳性对照组低4个数量级,4种酚类化合物的雌激素活性强弱顺序为辛基酚双酚A壬基酚2,4-二氯酚。可见金鱼可以作为类雌激素化合物筛选和检测的敏感实验生物。     

镉暴露对文蛤和四角蛤蜊血细胞功能及DNA损伤影响

为研究镉暴露对文蛤和四角蛤蜊血细胞功能及DNA损伤水平的影响,在实验室条件下将文蛤和四角蛤蜊暴露于镉离子浓度分别为0.05 mg·L~(-1)、0.10 mg·L~(-1)和0.25 mg·L~(-1)的海水中,用流式细胞仪测定两者血细胞的死亡率和活性氧含量,用溶酶体荧光探针和碱解旋法测定其溶酶体含量和DNA损伤。结果显示,在0.05 mg·L~(-1)暴露组文蛤和四角蛤蜊血细胞死亡率、活性氧含量分别从第3 d和第1 d开始明显高于对照组,在0.10 mg·L~(-1)暴露组均从实验开始即显著高于对照组,而在0.25 mg·L~(-1)暴露组两种蛤血细胞死亡率随时间一直明显上升,但四角蛤蜊活性氧含量在后期明显下降;0.05 mg·L~(-1)暴露组文蛤和四角蛤蜊溶酶体含量分别从第3 d和第1 d开始低于对照组,在其它浓度组从第1 d就低于对照组;文蛤和四角蛤蜊血细胞DNA F值在0.05 mg·L~(-1)暴露组分别从第10 d和第3 d开始明显低于对照组,文蛤和四角蛤蜊在0.10 mg·L处理组DNA F值分别从第3 d和第1 d开始明显低于对照组,且随时间下降,在0.25 mg·L~(-1)暴露组两者从第1 d开始即明显低于对照组。结果证实,文蛤和四角蛤蜊在高浓度镉离子影响下,受到的损伤较为严重,且部分指标在同浓度镉离子影响下,四角蛤蜊各指标变化要明显早于文蛤,短时间的0.05 mg·L~(-1)浓度镉离子暴露对文蛤影响不大,在长时间或高浓度镉离子暴露条件下,其指标变化幅度相比较四角蛤蜊也较小,所以四角蛤蜊对镉离子的敏感性更明显,其受到的威胁也较严重,是更适合的生物指示种,相关指标变化规律能恰当的体现镉对两种蛤的威胁程度。     

三丁基锡对文蛤外套膜代谢酶活性的影响

用实验生态学方法，观察浓度为0．1、1、10ng／dm^3的三丁基锡暴露2、8、20d以及恢复7、20d后对文蛤外套膜酸性磷酸酶、碱性磷酸酶以及Na^＋、K^＋-ATP酶活性的影响．结果表明各浓度组的三丁基锡在暴露早期（2d），外套膜中酸性、碱性磷酸酶和Na^＋、K^＋-ATP酶活性与对照组比较没有显著差异，随着暴露时间的延长，高浓度的TBT暴露对AKP表现出激活作用，对ACP和Na^＋、K^＋-ATP酶活性则具有抑制作用．结果显示Na^＋、K^＋-ATP酶可作为有机锡暴露的生物标志物．     

雌二醇、壬基酚和三丁基锡对斑马鱼雌激素响应基因表达的影响

为研究天然雌激素、环境拟雌激素和环境抗雌激素共存条件下的雌激素效应,分别以雌二醇(17β-estradiol, E2)、壬基酚(Nonylphenol, NP)和三丁基锡(Tributyltin, TBT)为代表,采用实时荧光定量PCR和相对定量分析法,测试并分析了3种污染物单独和不同组合联合暴露条件下成年雄性斑马鱼(Danio rerio)肝脏组织中雌激素响应基因vtg1、esr1、esr2a和esr2b mRNA的相对表达情况。结果显示:E2和NP的联合暴露,会显著诱导vtg1和esr1 mRNA的表达,且其相对表达量远高于两者单独暴露时的表达量之和。E2和TBT的联合暴露中,当TBT浓度较高(100 ng/L)而E2浓度较低(10 ng/L)时,TBT能够抑制E2对vtg1和esr1 mRNA表达的诱导作用。NP和TBT的联合暴露,会导致vtg1和esr1 mRNA的相对表达量远小于两者单独暴露时的表达量之和。而在E2、NP和TBT三者共同暴露中,当TBT浓度较高(100 ng/L),且E2和NP浓度较低(E2 10 ng/L, NP 25μg/L)时,TBT能抑制E2和NP对vtg1 mRNA表达的诱导作用。由此可见,不同类型环境雌激素共存条件下的联合效应与各组分的浓度配比有关,本研究结果为探索多种环境雌激素类污染物共存条件下的生物效应和生态风险提供了科学依据。     

饥饿对3种不同规格文蛤耗氧率和排氨率的影响

利用实验生态学的方法研究了饥饿对3种不同规格文蛤(Meretrix meretrix Linnaeus)耗氧率和排氨率的影响.实验饥饿的时间分别为1,3,5,7,10,13,20,27,35 d.实验结果表明,饥饿对3种不同规格的文蛤耗氧率和排氨率都有极其显著的影响(P<0.01).3种不同规格的文蛤在饥饿1～7 d时,耗氧率随着时间的延长而增大;饥饿7 d以后,耗氧率随着时间的延长而下降.3种不同规格的文蛤在饥饿1～10 d时,排氨率随着时间的延长而增大;在饥饿10 d以后,耗氧率随着时间的延长而下降.在实验周期内,大、中、小3种不同规格的文蛤的O/N值分别为15.05～36.12(平均23.69),15.6～37.56(平均24.73),17.44～47.25(平均27.17).     

瓦氏黄颡鱼(Pelteobagrus vachelli)卵黄蛋白原的纯化、性质鉴定及ELISA检测方法的建立

腹腔注射17β-雌二醇(E2),使瓦氏黄颡鱼雄鱼在7天内产生卵黄蛋白原(Vtg)。采用凝胶过滤和离子交换两种层析技术,从E2诱导的雄性瓦氏黄颡鱼血浆中分离、纯化出Vtg,采用糖、磷、脂蛋白染色技术证明分离、纯化的蛋白为Vtg,该Vtg在非变性条件下分子量约为240kDa,在SDS变性条件下分子量约为143kDa。纯化的瓦氏黄颡鱼Vtg经检测显示可能含有类胡萝卜素,但没有二硫键,对热相对稳定。利用纯化的瓦氏黄颡鱼Vtg,制备了兔抗瓦氏黄颡鱼Vtg多克隆抗血清。用双向免疫扩散法测得抗血清的纯度较高,效价为1︰32;Western blotting检测显示抗血清的特异性较好。以瓦氏黄颡鱼Vtg多克隆抗血清为抗体,以纯化的瓦氏黄颡鱼Vtg为抗原,建立了间接酶联免疫吸附反应(ELISA)方法检测瓦氏黄颡鱼体内Vtg的含量,标准曲线线性部分的线性方程为y=0.099x+0.4529(R2=0.9327),该方法检测的灵敏度为15.6ng/ml,工作范围为31.2—4000ng/ml,在此范围内,标准曲线具有良好的线性。     

四种重金属对刺参幼参的急性致毒效应

采用静水试验法,在水温18.5℃~20℃的条件下,用Cu2 ,Zn2 ,Cd2 和Cr6 四种重金属对刺参幼参进行了单一急性毒性试验。结果表明:Cu2 对刺参幼参的24h,48h,72h和96hLC50分别为0.299,0.176,0.133和0.120mg/L;Zn2 的24h,48h,72h和96hLC50分别为6.700,3.624,2.577和1.951mg/L;Cd2 的24h,48h和72hLC50分别为4.246,2.588和2.137mg/L;Cr6 的24h,48h和72hLC50分别为31.974,7.499和3.808mg/L;Cu2 ,Zn2 ,Cd2 和Cr6 对刺参幼参的安全浓度分别为0.018,0.362,0.259和0.750mg/L。四种重金属对刺参幼参的毒性大小依次为Cu2 >Cd2 >Zn2 >Cr6 。     

Tracing dissolved organic matter cycling in the eastern boundary of the temperate North Atlantic using absorption and fluorescence spectroscopy

Apparent oxygen utilization (AOU), dissolved organic carbon (DOC), coloured dissolved organic matter (CDOM) absorption spectra, and CDOM fluorescence characteristic of aromatic amino acids (excitation/emission 280 nm/320 nm; F(280/320)) and marine-humic like substances (320 nm/410 nm; F(320/410)) were measured in full depth during a cruise in the temperate Eastern North Atlantic (ENA). An optimum multi-parameter (OMP) inverse method was run to calculate water mass proportion-weighted average (archetypal) concentrations of these chemical parameters for all water masses and samples. Archetypal concentrations retain the variability due to water mass mixing and basin scale mineralization from the water mass formation sites to the study area. Conversely, the difference between measured and archetypal concentrations, retain the variability due to dissimilarities in mineralization processes within the study area. Our analysis indicates that DOC supported 26±3% of the AOU in the dark temperate ENA and that basin scale processes occurring at and from the formation area of the water masses explained 63% of the total DOC variability. Our data also suggests that DOC remineralized at the basin scale was of lower molecular weight, and with a lower proportion of fluorescent aromatic amino acids than found within the study area. The relationship between the absorption coefficient at 254 nm (a_CDOM(254)) and AOU indicates that a_CDOM(254) was consumed during organic matter remineralization in the dark ocean, with 55% of the variability being explained by basin scale processes. The relationships of F(320/410) with AOU and DOC confirmed that marine humic-like substances are produced by microbial degradation processes, at a rate of 6.1±0.9×10⁻³ mg equivalents of QS mol AOU⁻¹. Our results also indicate that basin-scale remineralization processes account for 85% of the total variability of F(320/410), emphasizing that large scale processes control the formation of humic-like substance in the dark ENA.     

海洋中β-二甲基巯基丙酸内盐降解过程的研究进展

二甲基硫(DMS)是海水中一种最重要的、含量最丰富的还原态挥发性生源有机硫化物,前体β-二甲基巯基丙酸内盐(DMSP)的降解过程受各种因素影响。其中主要包括温度、DMSP的浓度、氧气、盐度、酸度、颗粒粒度、藻类生长期、季节变化、氧化压力、抑制剂等。它们均与DMSP降解速率呈一定的函数关系,并对DMSP的降解产物产生影响。藻类是DMSP的主要来源,因此着重讨论了温度、盐度、酸度等对不同浮游植物细胞内DMSP与DMS生物生产和转化过程的影响。结合海洋硫循环的研究现状和海洋化学发展的趋势,探究了用颗粒态DMSP与Chla的比率来量化碳和硫通量的方法及DMSP裂解酶活性的检验技术。大气中CO2压力持续增加导致的海洋酸化对藻类中DMSP降解过程的影响也是进一步研究的重点。     

Ballast water management in Canada: A historical perspective and implications for the future

Non-indigenous species (NIS) can create significant risks when introduced to foreign ecosystems. One of the many vectors facilitating the global transport of NIS is ballast water, and as commercial shipping increases so does the threat of ballast water-mediated introductions. Despite Canadian regulations, consistent with international guidelines put forth by the International Maritime Organization in the early 1990s, ballast water remains a potential vector for the introduction of many NIS to Canadian marine ecosystems: there have been no new introductions in the Laurentian Great Lakes Region since 2006, yet marine ecosystems continue to see the introduction and establishment of NIS. This paper details and analyzes the development of Canadian ballast water management (BWM), in the context of marine ecosystems, and identifies issues therein. BWM in Canada has a number of management gaps that have persisted since large-scale BWM began in the late 1980s. These include BWM exemptions for vessels not exiting the North American continental shelf, and limitations of effectiveness of mid-ocean exchange as the predominant management method in marine ecosystems. In addition, ballast water regulations for the Canadian Arctic may require additional consideration as the Arctic will likely continue to see an increase in both warming and commercial shipping in the future, thereby increasing the potential risk of NIS. In order to adequately protect all Canadian marine ecosystems, it is recommended that the current BWM regime: fill gaps in management to help prevent further introduction and spread of NIS; focus equally on both marine and freshwater regions, and; look to other aggressive BWM strategies such as those being developed by the state of California.     

Sources and implications of particulate organic matter from a small tropical river——Zuari River,India

Transitional ecosystems,estuaries and the coastal seas,are distinctively affected by natural and anthropogenic factors.Organic matter(OM)originating from terrestrial sources is exported by rivers and forms a key component of the global biogeochemical cycles.Most previous studies focused on the bulk biochemical and anthropogenic aspects affecting these ecosystems.In the present study,we examined the sources and fate of OM entrained within suspended particulate matter(SPM)of the Zuari River and its estuary,west coast of India.Besides using amino acid(AA)enantiomers(L-and D-forms)as biomarkers,other bulk biochemical parameters viz.particulate organic carbon(POC),δ13C,particulate nitrogen(PN),δ15N and chlorophyll a were analyzed.Surprisingly no significant temporal variations were observed in the parameters analyzed;nonetheless,salinity,POC,δ13C,PN,δ15N,glutamic acid,serine,alanine,tyrosine,leucine and D-aspartic acid exhibited significant spatial variability suggesting source differentiation.The POC content displayed weak temporal variability with low values observed during the post-monsoon season attributed to inputs from mixed sources.Estuarine samples were less depleted than the riverine samples suggesting contributions from marine plankton in addition to contributions from river plankton and terrestrial C3 plants detritus.Labile OM was observed during the monsoon and post-monsoon seasons in the estuarine region.More degraded OM was noticed during the pre-monsoon season.Principal component analysis was used to ascertain the sources and factors influencing OM.Principally five factors were extracted explaining 84.52%of the total variance.The first component accounted for 27.10%of the variance suggesting the dominance of tidal influence whereas,the second component accounted for heterotrophic bacteria and their remnants associated with the particulate matter,contributing primarily to the AA pool.Based on this study we ascertained the role of the estuarine turbidity maximum(ETM)controlling the sources of POM and its implications to small tropical rivers.Thus,changes in temporal and regional settings are more likely to affect the natural biogeochemical cycles of small tropical rivers.     

Seabed Ground Improvement in Coastal Reclamation Area by Heavy Tamping After Rockfilling Displacement

Considering the characteristics of seabed ground in coastal reclamation area, ultra soft clay improvement method by heavy tamping after rockfilling displacement was proposed combined with a case study. The improvement mechanisms of the method can primarily be attributed to rockfilling displacement (RD), dynamic compaction (DC), dynamic replacement (DR) and dynamic replacement and mixing (DRM). For the case study given in this article, heavy tamping construction program was proposed based on field pilot tests. Furthermore, the effectiveness of the proposed ground improvement method was verified through in situ plate load test, sand fill test and the analyses of observed settlement data. Thus, the method of heavy tamping after rockfilling displacement is applicable for the improvement of seabed ground in coastal reclamation area. It is proposed for similar projects that heavy tamping of fills may be performed by layers and correspondingly tamping energy increased to further enhance the improvement effect of DC, DR and DRM. In addition, delayed improvement effect or time effect of soil mass after heavy tamping is still an issue to be further studied.     

莱州湾海域水体中有机氯农药和多氯联苯的浓度水平和分布特征

以固相萃取/气相色谱法测定了莱州湾海域水体中22种有机氯农药和多氯联苯类化合物的浓度水平和分布特征。结果表明,莱州湾海域表层水体中有机氯农药浓度范围为N.D.~32.7ng/L,底层水中的浓度范围为N.D.~11.7ng/L。在该海域水体中共检出有机氯农药3种,β-666是水体中主要的有机氯农药污染物。多氯联苯类在底层水样中检出2种,总浓度范围在4.5~27.7ng/L之间。该海域有机氯农药和多氯联苯的分布特征是近岸高,离岸低,由近岸向湾外延伸方向依次递减。并对莱州湾表层水中总有机氯农药与海水盐度、氯度、溶解氧和pH间的关系作了初步探讨,得出总有机氯农药与盐度、氯度间有一定的相关关系,相关系数均为0.59。方法测定5种有机氯农药化合物的空白加标回收率为97.3%~126.0%,相对标准偏差为2.8%~8.6%;测定5种多氯联苯类化合物的空白加标回收率为88.6%~151.8%,相对标准偏差为6.7%~10.4%。     

空间信息系统原理(三)数字高程模型及其数据结构

简要介绍了DTM的提出和应用情况,并对数字地面模型(DTM)、数字高程模型(DEM或DHM)、数字地形模型(DGM)等三个概念进行了论述,说明了它们的区别和联系,详细论述了离散点、不规则三角形、等高线、断面线和规则格网等几种数字高程模型的数据结构,并对不同数据结构的数字地面模型的特征进行了比较。     

Material selection for the gas containment system of a compressed natural gas carrier fleet

In this work, the possible exploitation of fiber-reinforced composites in the context of maritime transportation of compressed natural gas (CNG) is investigated. In addition to a more conventional steel configuration, two different fiber materials, carbon and glass, are considered as construction materials for pressure vessels (PVs) to be stored on board ships, with thickness optimized by FEM analysis.The considered scenario is represented by the transportation of CNG from an offshore well to a terminal on shore. Fleets of ships carrying CNG in pressure vessels manufactured with the investigated materials are generated by means of a ship synthesis model (SSM) software and compared on the basis of technical and economical indicators.The choice of the construction material influences considerably the weight of the PVs, which represent a major item of total ship weight and reflects directly on the general transport performances in terms of resistance, seakeeping and reliability in the service. On the other hand, capital as well as operating expenditures are considerably affected by the choice. When exploring the design space, the ship synthesis model is able, at a preliminary stage of the design, to account for the various technical and economical aspects, their implications and relationships. Results are presented of computations carried out in a specific case, identified by the annual gas production and other characteristics of the well terminal and a cruising route for the ships. The comparison is carried out on the basis of the cost per transported unit of gas and of the percentage of success in the transportation process. The computations show that the choice of the PV material has a key influence on the results in terms of optimal number, dimensions and speed of the ships.     

基于EMD与神经网络的机械故障诊断技术

经验模式分解 (EMD)是分析非线性、非平稳信号的有力工具 ,它将信号分解为突出了原信号的不同时间尺度的局部特征信息的内在模函数 (IMF)分量。本文通过将各 IMF分量输入到 BP网络中进行训练学习和故障诊断 ,比直接输入原信号可以提高 BP网络对故障诊断的准确率 ,而且减少了训练时间。