A fluorescence-based bioassay for aquatic macrophytes and its suitability for effect analysis of non-photosystem II inhibitors

Background, Goals and Scope During the last years the miniaturization of toxicity test systems for rapid and parallel measurements of large quantities of samples has often been discussed. For unicellular algae as well as for aquatic macrophytes, fluorescence-based miniaturized test systems have been introduced to analyze photosystem II (PSII) inhibitors. Nevertheless, high-throughput screening should also guarantee the effect detection of a broad range of toxicants in order to ensure routinely applicable, high-throughput measuring device experiments which can cover a broad range of toxicants and modes of action others than PSII inhibition. Thus, the aim of this study was to establish a fast and reproducible measuring system for non-PSII inhibitors for aquatic macrophyte species to overcome major limitations for use. Methods A newly developed imaging pulse-amplitude-modulated chlorophyll fluorometer (I-PAM) was applied as an effect detector in short-term bioassays with the aquatic macrophyte species Lemna minor. This multiwell-plate based measuring device enabled the incubation and measurement of up to 24 samples in parallel. The chemicals paraquat-dichloride, alizarine and triclosan were chosen as representatives for the toxicant groups of non-PSII herbicides, polycyclic aromatic hydrocarbons (PAHs) and pharmaceuticals and personal care products (PPCPs), which are often detected in the aquatic environment. The I-PAM was used (i) to establish and validate the sensitivity of the test system to the three non-PSII inhibitors, (ii) to compare the test systems with standardized and established biotests for aquatic macrophytes, and (iii) to define necessary time scales in aquatic macrophyte testing. For validation of the fluorescence-based assay, the standard growth test with L. minor (ISO/DIS 20079) was performed in parallel for each chemical. Results The results revealed that fluorescence-based measurements with the I-PAM allow rapid and parallel analysis of large amounts of aquatic macrophyte samples. The I-PAM enabled the recording of concentration-effect-curves with L. minor samples on a 24-well plate with single measurements. Fluorescence-based concentration-effect-curves could be detected for all three chemicals after only 1 h of incubation. After 4–5 h incubation time, the maximum inhibition of fluorescence showed an 80–100% effect for the chemicals tested. The EC50 after 24 h incubation were estimated to be 0.06 mg/L, 0.84 mg/L and 1.69 mg/L for paraquatdichloride, alizarine and triclosan, respectively. Discussion The results obtained with the I-PAM after 24 h for the herbicide paraquat-dichloride and the polycyclic aromatic hydrocarbon alizarine were in good accordance with median effective concentrations (EC50s) obtained by the standardized growth test for L. minor after 7 d incubation (0.09 mg/L and 0.79 mg/L for paraquat-dichloride and alizarine, respectively). Those results were in accordance with literature findings for the two chemicals. In contrast, fluorescence-based EC50 of the antimicrobial agent triclosan proved to be two orders of magnitude greater when compared to the standard growth test with 7 d incubation time (0.026 mg/L) as well as with literature findings. Conclusion Typically, aquatic macrophyte testing is very time consuming and relies on laborious experimental set-ups. The I-PAM measuring device enabled fast effect screening for the three chemicals tested. While established test systems for aquatic macrophytes need incubation times of ≥ 7 d, the I-PAM can detect inhibitory effects much earlier (24 h), even if inhibition of chemicals is not specifically associated with PSII. Thus, the fluorescence-based bioassay with the I-PAM offers a promising approach for the miniaturization and high-throughput testing of chemicals with aquatic macrophytes. For the chemical triclosan, however, the short-term effect prediction with the I-PAM has been shown to be less sensitive than with long-term bioassays, which might be due to physicochemical substance properties such as lipophilicity. Recommendations and Perspectives The results of this study show that the I-PAM represents a promising tool for decreasing the incubation times of aquatic macrophyte toxicity testing to about 24 h as a supplement to existing test batteries. The applicability of this I-PAM bioassay on emergent and submerged aquatic macrophyte species should be investigated in further studies. Regarding considerations that physicochemical properties of the tested substances might play an important role in microplate bioassays, the I-PAM bioassay should either be accompanied by evaluating physicochemical properties modeled from structural information prior to an experimental investigation, or by intensified chemical analyses to identify and determine nominal concentrations of the toxicants tested. The chemicals paraquat-dichloride, alizarine and triclosan were chosen as representatives for the toxicant groups of non-PSII herbicides, PAHs and PPCPs which are often detected in the aquatic environment. Nevertheless, in order to ensure a routinely applicable measuring device, experiments with a broader range of toxicants and samples of surface and/or waste waters are necessary. ESS-Submission Editor: Dr. Markus Hecker (MHecker@Entrix.com)     

新疆矿区煤矿瓦斯排放现状与趋势分析

为获得新疆矿区当前生产规模对应的瓦斯排放基量,调查了研究矿区生产矿井的瓦斯排放现状.根据各矿区2004-2006年生产矿井的开采煤层、生产水平、生产规模和瓦斯排放等参数,对不同矿区和煤田进行了统计分析,计算了新疆矿区当前生产规模对应的瓦斯排放总量,同时对重点瓦斯防治矿区和一般瓦斯防治矿区进行了分类.调查结果表明,新疆矿区煤矿平均吨煤相对瓦斯涌出量由2004年的2.82m3/t增加到2006年的3.09 m3/t,呈逐年递增趋势; 新疆矿区2004-2006年的年平均瓦斯(CH4)排放量为1.05×108 m3/a;准南煤田(I)和塔北煤田(VI)的平均绝对瓦斯涌出量分别占全疆绝对瓦斯涌出总量的64.82%和15.19%,为相对瓦斯富集区,也是当前瓦斯防治的重点区域.按照矿区3年平均绝对瓦斯涌出量(大于1.00m3/min)或平均相对瓦斯涌出量(大于5m3/t)将新疆矿区分为重点瓦斯防治矿区和一般瓦斯防治矿区,并提出了瓦斯分类治理的具体建议.应用灰色系统理论,结合调查数据分别求解了新疆矿区原煤产量与绝对瓦斯涌出量的GM[1,1]预测模型.本研究为新疆煤炭资源后续规模开发过程中的相关环境决策和节能减排政策的实施提供了依据.     

飞行安全的针对性评估

根据飞行员执行新的或特殊任务的需要,在研制飞行安全评估系统时,设置了针对性评估模块.运用层析分析法(AHP)确定了针对性评估模块与一般性评估模块的权重关系.根据飞行安全理论确定了一般性评估简捷方法的因素、要素及量化数据;根据层析分析法、基元事件分析法确定了针对性评估模块的结构和因素的权重,采用基元事件分析法、飞行法规分析和调查研究相结合的方法,对针对性评估的要素进行了量化.最后针对具体事例做了评估计算.研究表明,当针对性评估的安全度小于90%时,应引起足够的警觉,采取必要的措施.     

危险品道路运输泄漏引发水污染事故的定量风险评估方法研究

介绍了由道路交通事故引发的地表水污染事故风险定量分析方法,重点分析了突发水污染事故源发生概率和危害后果表征方法.以评价水域内道路危险品运输泄漏事故发生概率为水污染事故源发生概率.污染事故后果危害水平的表征:1)敏感区(点)最高浓度; 2)敏感区(点)超标总历时;3)水质超标的范围(如超标河流长度).评估方法为陆源造成的突发水污染事故的预防管理与应急计划的制订实施提供了科学依据.     

基于人工神经网络理论的建筑物火灾安全评价研究

依据建筑物火灾危险性的影响因素,应用人工神经网络理论及系统安全方法,建立了建筑物火灾危险性的评价指标体系,该方法摆脱了评价过程中的随机性和参评人员主观上的不确定性及其认识上的模糊性等缺点,大大提高了准确性。为了验证评价模型的准确性,将该理论应用到某高校图书馆火灾危险性评价中,快速、准确地得到了安全评价结果,取得了满意效果,为建筑物防火设计以及安全管理提供了可行的依据。     

冲压机械伤害风险评价信息系统研究

研究冲压机械伤害风险评价信息系统,无论对企业内部事故预防还是对职工健康安全,还是节省国家因事故造成的经济损失,都十分必要。应用计算机编程语言和数据库软件,对冲压作业进行综合管理评价、设备设施的固有危险性评价及劳动卫生与作业环境评价,进而对冲压作业风险进行评价分级,建立动态的冲压机械伤害风险评价数据库管理信息系统。该系统可保存所有评价的项目资料,供应用和参考,实现了对冲压机械伤害风险评价的数据库动态管理,使风险评价分析工作组织化、条理化、系统化。     

企业安全文化评估与企业安全行为的质化研究

通过总结国内外各种不同的安全文化概念,归纳分析了国内外较为流行的近10种安全文化建立模式。在研讨文献的基础上,通过深入访谈、焦点小组与参与观察,归纳构造了一套共11个维度,总题项为152题的安全文化评估量表。通过对选定企业的实证问卷调查,进行统计分析,采用多元逐步回归分析等方法对安全文化各评核因素及整体安全文化进行预测,并以百分数呈现其整体的安全文化现状水平。通过基于扎根理论的各种质化研究方法,对案例企业的安全文化建设和企业安全行为进行研究,得到了7点关于企业安全行为的研究发现,并提出了相应的改进建议。     

安全评价方法在危险废物处置环境风险评价中的应用探讨

对安全评价方法在危险废物处置建设项目环境风险评价中的运用进行初步探索。主要用"工艺过程风险因素分析表"对工艺过程潜在风险性识别;用蒙德法进行源项分析;用池火灾模型、蒸气云爆炸伤害模型对易燃、易爆物质的火灾、爆炸等重大事故后果进行计算,得出人员的伤亡半径和财产损失半径等参数,以便于判断风险的可接受水平。分析结果表明:采用安全评价方法对危险废物处置建设项目进行环境风险评价是适用的、可行的。     

“四沿”化工企业污水处理系统安全事故状态下负荷缓冲及安全环境(SE)联锁探讨

在对"四沿"(沿江、沿河、沿湖、沿海)化工行业污水处理方法及流程特点总结的基础上,归纳出应对水力和污染物冲击负荷的缓冲方法。为克服缓冲控制能力有限的缺点,提出一种新方法——SE(安全和环境)联锁,以期达到完善管理、提高控制效果的目的,并详细阐述了该方法在管理及技术层面的实施途径。     

油气集输站安全监控预警及管理信息系统的开发

提出油气集输站安全监控预警及管理信息系统建设的调研内容、需求分析以及安全相关控制系统的风险分析流程。设计了该系统建设的4层结构框架和拓扑图,深入研究了实时参数监控、分级报警与安全保护控制、智能诊断与预警、多媒体监控、安全生产管理信息子系统的主要开发内容和功能实现。