Toxicity assessment of a common laundry detergent using the freshwater flagellate Euglena gracilis

Synthetic detergents are among the commonly used chemicals in everyday life. Detergents, reaching aquatic environments through domestic and municipal wastewater, can cause many different effects in aquatic organisms. The present study was aimed at the toxicity evaluation of a commonly used laundry detergent, Ariel, using the freshwater flagellate Euglena gracilis as a biotest organism. Different parameters of the flagellate like motility, swimming velocity, cell shape, gravitactic orientation, photosynthesis and concentration of light harvesting pigments were used as end points for the toxicity assessment. No Observed Effect Concentration (NOEC) and EC₅₀ values were calculated for the end point parameters at four different incubation times, i.e. 0, 6, 24 and 72 h. After 72 h incubation, swimming velocity of the cells was found to be the most sensitive parameter giving NOEC and EC₅₀ values of 10.8 and 34 mg L⁻¹, respectively. After 72 h exposure to the detergent, chlorophyll a and total carotenoids were significantly decreased in cultures treated with Ariel at concentrations of 50 mg L⁻¹ and above while chlorophyll b significantly decreased at concentrations above 750 mg L⁻¹. The maximum inhibitory effect on the quantum yield of photosystem II was observed after 24 h exposure and thereafter a recovery trend was observed. Motility, gravitaxis and cell shape were strongly impaired immediately upon exposure to the detergent, but with increasing exposure time these parameters showed acclimatization to the stress and thus the NOEC values obtained after 72 h were higher than those immediately after exposure.     

Assessing benzene-induced toxicity on wild type Euglena gracilis Z and its mutant strain SMZ

Benzene is a representative member of volatile organic compounds and has been widely used as an industrial solvent. Groundwater contamination of benzene may pose risks to human health and ecosystems. Detection of benzene in the groundwater using chemical analysis is expensive and time consuming. In addition, biological responses to environmental exposures are uninformative using such analysis. Therefore, the aim of this study was to employ a microorganism, Euglena gracilis (E. gracilis) as a putative model to monitor the contamination of benzene in groundwater. To this end, we examined the wild type of E. gracilis Z and its mutant form, SMZ in their growth rate, morphology, chlorophyll content, formation of reactive oxygen species (ROS) and DNA damage in response to benzene exposure. The results showed that benzene inhibited cell growth in a dose response manner up to 48 h of exposure. SMZ showed a greater sensitivity compared to Z in response to benzene exposure. The difference was more evident at lower concentrations of benzene (0.005–5 μM) where growth inhibition occurred in SMZ but not in Z cells. We found that benzene induced morphological changes, formation of lipofuscin, and decreased chlorophyll content in Z strain in a dose response manner. No significant differences were found between the two strains in ROS formation and DNA damage by benzene at concentrations affecting cell growth. Based on these results, we conclude that E. gracilis cells were sensitive to benzene-induced toxicities for certain endpoints such as cell growth rate, morphological change, depletion of chlorophyll. Therefore, it is a potentially suitable model for monitoring the contamination of benzene and its effects in the groundwater.     

三种类型油菜(Brassica spp.)和野芥菜(B. juncea var.gracilis Tsen et Lee)杂交亲和性及F1的适合度--潜在基因转移的研究

通过人工去雄授粉，采用荧光显微镜观察了3种类型栽培油菜花粉在两地采集的野芥菜柱头上的萌发生长情况，结合杂交后的结实率，探讨了3种类型油菜和野芥菜杂交的亲和性．结果表明，甘蓝型油菜和芥菜型油菜与野芥菜的亲和性都非常高，亲和性指数达10．0以上，而白菜型油菜和野芥菜的亲和性较低，亲和指数小于0．2．子一代的适合度研究结果表明，芥菜型油菜向野芥菜杂交一代的适合度没有降低，而甘蓝型及白菜型和野芥菜杂交一代的适合度明显下降，表现在花粉活力降低，结实率极低．上述结果表明，白菜型油菜和野芥菜的基因转移可能性最小，甘蓝型居中，而芥菜型极易向野芥菜发生基因转移．     

Euglena gracilis as a model for the study of Cu and Zn toxicity and accumulation in eukaryotic cells

We have observed the effect of copper and zinc on the biology of Euglena gracilis. The cells displayed different sensitivities to these metals, as the apparent LC₅₀ for Cu²⁺ was 0.22 mM, and for Zn²⁺ it was 0.88 mM. While Zn²⁺ was able to increase cell proliferation even at 0.1 mM, the minimal CuCl₂ concentration tested (0.02 mM) was sufficient to impair cell division. Higher concentrations of these metals not only inhibited cell division in a concentration-dependent manner, but also interfered with the metabolism of E. gracilis. A higher accumulation of proteins and lipids per cell was observed at the DI₅₀ concentration for metal-treated cells. These results suggest that the test concentration of both metals leads to a failure in completing cell division. Ultrastructural analysis indicated a chloroplast disorganization in copper-treated cells, as well as the presence of electron dense granules with different shapes and sizes inside vacuoles. Microanalysis of these granules indicated an accumulation of copper, thus suggesting a detoxification role played by the vacuoles. These results indicate that E. gracilis is an efficient biological model for the study of metal poisoning in eukaryotic cells. They also indicate that copper and zinc (copper being more poisonous) had an overall toxic effect on E. gracilis and that part of the effect can be ascribed to defects in the structure of chloroplast membranes.     

桉树叶水浸出液对2种海洋微藻生长和叶绿素荧光特性的影响

运用水样叶绿素荧光仪(Water-PAM)研究了桉树叶水浸出液胁迫下湛江叉鞭金藻(Dicrateria zhanjiangensis)和纤细角毛藻(Chaetoceros gracilis)2种海洋微藻叶绿素荧光特性〔包括PS(光系统)Ⅱ的F_v/F₀(潜在活性)、F_v/F_m(最大光能转化效率)、Yield(实际光能转化效率)、ETR(电子传递效率),其中F_m和F₀分别为最大荧光和基础荧光〕的变化,同时监测微藻的生长情况,以研究桉树叶水浸出液对海洋微藻的影响,从而为探讨华南沿海大量种植桉树所造成潜在的海洋生态问题提供一些依据. 结果表明:①低浓度(以φ计,0.4、0.8、1.2mL/L)胁迫在96h内对湛江叉鞭金藻的上述4项荧光参数均无显著影响(P>0.05);高浓度(2.0mL/L)胁迫下,24h时湛江叉鞭金藻的4项荧光参数都处于最低值,48h时处于次低值. 表明高浓度在胁迫早期(48h内)对湛江叉鞭金藻4项荧光参数有抑制作用,而后期(72、96h)这4项荧光参数又恢复到正常水平,抑制作用解除. ②低浓度(0.4、0.8mL/L)胁迫下纤细角毛藻的4项荧光参数在24、48h内均得到促进,而高浓度(2.0mL/L)胁迫下F_v/F_m在24h时为最小值,表现抑制作用;另外3项荧光参数则未受显著影响. 在胁迫后期(72、96h),纤细角毛藻的4项荧光参数均随着胁迫浓度的增大而增加,表现为促进作用. ③就生长情况而言,随着胁迫浓度的增大,2种海洋微藻细胞密度的增长明显变缓. 表明桉树叶水浸出液对2种海洋微藻的生长以及叶绿素荧光特性都有影响,从而将会对整个海洋生态系统产生影响.      

太湖梅梁湾有机污染物对纤细裸藻（Euglena gracilis）的遗传毒性效应

采用GC-MS方法分析了太湖梅梁湾水样的主要有机污染物成分和含量,共检测出15种目标化合物,表明该区域水体已经受到有机物污染;纤细裸藻(Euglena gracilis)在低浓度浓缩水样处理下(1倍组)生长无显著影响,叶绿素a、b含量和类胡萝卜素含量比对照组显著增加,而高浓度下(5倍和10倍组)生长明显受到抑制,叶绿素a和类胡萝卜素含量呈下降趋势;抗氧化酶系统中SOD和POD活性呈上升趋势,显示有机污染物胁迫可诱导抗氧化酶活性;彗星试验中,Olive尾矩和尾动量增加,纤细裸藻细胞DNA损伤程度随着太湖水样污染物浓度增加而加重,呈现出明显的剂量-效应关系,提示太湖梅梁湾水样具有潜在致突变性.结果表明,慧星试验和SOD活性试验结合使用在水环境中的遗传毒性监测方面具有较大的应用价值,适合作为水体有机污染监测的生物标志物.     

应用模式生物检测饮用水源颗粒物的毒性效应

悬浮颗粒物在自然水体中广泛存在,大量小颗粒物(粒径<1μm)可吸附、聚集水中多种污染物,从而形成新的复合污染,影响水生生态系统。已有研究表明,颗粒物粒径越小,传统饮用水处理工艺越难以去除,因此,其表面吸附的污染物也会随之残留在水体中,对饮用水安全产生影响。本研究对南京某自来水厂进水和出水中的颗粒物进行了测定和对比,并选用3种不同种类的模式生物包括发光菌(Photobacterium phosphoreum)、纤细裸藻(Euglena gracilis)和蚕豆(Vicia faba L.),对饮用水源水中不同粒径颗粒物的毒性效应进行了测试。结果表明,传统水处理工艺对小颗粒物(粒径<1μm)的去除效果并不理想;经传统水处理工艺处理后,出水仍显著抑制发光菌发光强度以及纤细裸藻生长,对蚕豆的污染指数较高,可能存在一定健康隐患;进水中颗粒物粒径<1μm组对发光菌发光强度和纤细裸藻生长的抑制效应显著大于粒径≥1μm颗粒物组,蚕豆根尖的污染指数也更高,毒性更强。本研究结果可为饮用水的生物毒性检测和水处理工艺改良提供参考。     

A new approach for biological online testing of stack gas condensate from municipal waste incinerators

A biological testing system for the monitoring of stack gas condensates of municipal waste incinerators has been developed using Euglena gracilis as a test organism. The motility, velocity and cellular form of the organisms were the endpoints, calculated by an image analysis system. All endpoints showed statistically significant changes in a short time when organisms were exposed to samples collected during combustion situations with increased pollutant concentrations. The velocity of the organisms proved to be the most appropriate endpoint. A semi-continuous system with E. gracilis for monitoring stack gas condensate is proposed, which could result in an online system for testing stack gas condensates in the future.     

高浓度氨氮胁迫对纤细裸藻的毒性效应

采用室内培养方法,通过检测纤细裸藻生长、光合色素含量、抗氧化酶活性及DNA损伤(彗星实验)研究了高浓度氨氮胁迫对纤细裸藻(Euglena gracilis)的毒性效应,以期为氨氮的水生态风险评价以及藻类污水净化提供科学依据.结果表明,氨氮在所设定的浓度范围内抑制藻类的生长,浓度越高,抑制越明显,2 000 mg·L-1时相比对照抑制率达55.7%;叶绿素含量随氨氮浓度增加先升高后下降,蛋白质含量与叶绿素变化趋势基本吻合;抗氧化酶系统中超氧化物歧化酶(SOD)和过氧化物酶(POD)活性随氨氮浓度增加而上升,2 000 mg·L-1时比对照分别增加了30.7%和49.4%,提示氨氮胁迫可诱导抗氧化酶活性增加;彗星实验中,纤细裸藻细胞DNA损伤程度随氨氮浓度增加而加重,表明高浓度氨氮具有潜在的致突变性.     

桉树与几种滨海植物水浸提液对纤细角毛藻生长的影响

为了研究桉树人工林是否会对海洋初级生产者产生影响,实验选取了桉树及4种滨海植物(木麻黄、台湾相思、秋茄和白骨壤),对比5种植物叶片水浸提液对纤细角毛藻生长的影响。结果显示,桉树对纤细角毛藻生长的影响最大,低浓度的叶片水浸提液(5 m L/L)表现出很强的抑藻作用;木麻黄、台湾相思和秋茄对纤细角毛藻生长的影响次之,低浓度的叶片水浸提液(5 m L/L)表现出较弱的抑藻作用,15 m L/L时,对藻的抑制作用较强;白骨壤对纤细角毛藻生长的影响最弱,25 m L/L时,表现出对藻有较弱的抑制作用。实验结果表明,桉树比其他常见的滨海植物对海洋微藻具有更强的化感作用,在沿海地区大面积种植桉树人工林会对海洋初级生产者产生潜在的影响。