)]发光细菌法在水质综合毒性在线检测中的应用

以海洋发光细菌费氏弧菌(Vibrio fischeri)作为检测生物,采用冻干菌粉快速复苏技术,研究费氏弧菌在水质检测中的最佳测试温度和有效性测试条件,并对硫酸锌等多种毒物和几种实际水样进行发光抑制作用分析。研究表明,费氏弧菌冻干粉复苏菌液保存在2~5℃条件下能有效测试7 d,最佳测试温度为15℃,最佳测试时间为15 min。氯化汞、硫酸锌、硫酸镉等重金属和苯胺、多菌灵、甲醛等有机毒物对费氏弧菌均具有较强的光抑制作用,也即费氏弧菌对以上毒物较为敏感,并能够连续7 d保持对同一浓度硫酸锌的敏感性较为一致。对几种实际水样的测试和分析表明,以费氏弧菌为指示生物的发光细菌法能够应用于水质环境安全的综合毒性在线监测预警中。     

Acute toxicity assessment of drinking water source with luminescent bacteria: Impact of environmental conditions and a case study in Luoma Lake,East China

• Acute toxicity assessment was conducted in Luoma lake watershed, East China. • Impacts of environmental factors on the toxicity testing was fully evaluated. • Dissolve oxygen had a weak positive correlation with luminescence inhibition rate. Protecting the quality of lake watersheds by preventing and reducing their contamination is an effective approach to ensure the sustainability of the drinking water supply. In this study, acute toxicity assessment was conducted on the basis of acute bioluminescence inhibition assay using the marine bacterium Vibrio fischeri as the test organism and Luoma Lake drinking water source in East China as the research target. The suitable ranges of environmental factors, including pH value, organic matter, turbidity, hardness, and dissolved oxygen of water samples were evaluated for the toxicity testing of bioluminescent bacteria. The physicochemical characteristics of water samples at the selected 43 sites of Luoma Lake watershed were measured. Results showed that the variations in pH value (7.31–8.41), hardness (5–20 °d) and dissolved oxygen (4.44–11.03 mg/L) of Luoma Lake and its main inflow and outflow rivers had negligible impacts on the acute toxicity testing of V. fischeri. The luminescence inhibition rates ranged from -11.21% to 10.80% at the 43 sites. Pearson’s correlation analysis in the experiment revealed that temperature, pH value, hardness, and turbidity had no correlation with luminescence inhibition rate, whereas dissolved oxygen showed a weak statistically positive correlation with a Pearson correlation coefficient of 0.455 (p<0.05).     

Screening organic micropollutants in surface waters by SPE extraction and ecotoxicological testing

Complex mixtures of toxic substances occurring in surface waters are difficult to characterise by chemical analyses because each compound occurs at a very low concentration and requires a specific analytical method to be identified. Ecotoxicological tests on water extracts can be used as a screening tool to evaluate quickly and simply the overall quality of a water body with regard to micropollutant contamination. In this work, a pre-concentration procedure based on solid-phase extraction (SPE), suitable for both biological testing and analytical determination, is proposed. The extraction procedure is an improved version of a methodology used to evaluate the toxicity of organic micropollutants occurring in surface waters. It offers the advantage of using disposable commercial cartridges, which are easier to manage than the columns prepared with macromolecular resins. Water extracts from two representative Italian rivers, characterised by a different gradient of potential contamination and prepared according to the new concentration techniques, are used. The acute toxicity of the water extracts is tested on Daphnia magna and the bioluminescence inhibition in Vibrio fischeri. Criteria based on the concentration factor (CF) are proposed for assessing the hazard to aquatic life due to the exposure to toxic substances in surface waters. The aim of hazard ranking is to focus analytical efforts towards those samples that show the highest toxic potential.     

Effects of a municipal effluent on the freshwater mussel Elliptio complanata following challenge with Vibrio anguillarum

The purpose of this study was to examine the cumulative effects of exposure to a pathogenic bacteria and municipal effluent in the freshwater mussel Elliptio complanata. Mussels were exposed to increasing concentrations of an ozone-treated effluent at 15°C for 7 days. A sub-group of mussels was inoculated with Vibrio anguillarum and exposed to the same conditions as above.After the exposure period, mussels were collected to assess hemocyte count and viability,immunocompetence(phagocytosis and nitrite production), oxidative stress/inflammation(cyclooxygenase and lipid peroxidation) and oxygen radical/xenobiotic scavenging activity(metallothioniens, glutathione S-transferase). The results showed that mussels exposed to municipal effluent had increased hemocyte counts, phagocytosis, nitrites, lipid peroxidation and metallothioneins. In the inoculated mussels, the same responses were observed, in addition to cyclooxygenase and glutathione S-transferase activities. Multivariate analyses revealed that(1)the response pattern changed with effluent concentration, where increased responses observed at low effluent concentrations(10%, V/V) were attenuated at higher effluent concentrations,(2)the effluent produced more pronounced changes in lipid peroxidation, metallothionein and hemocyte viability, and(3) the simultaneous presence of V. anguillarum led to more important changes in hemocyte count and viability and nitrite levels. In conclusion, the presence of V.anguillarum could alter the response of mussels to municipal effluent, which could lead to increased inflammation in mussels.     

磺胺与金属氧化物纳米颗粒对发光菌的急性联合毒性及其机制初探

磺胺与金属氧化物纳米颗粒同作为抗菌剂对环境与人体健康皆有危害。为探究其对细菌的毒性作用,测定了4种磺胺,包括磺胺氯哒嗪(SCP)、磺胺吡啶(SPY)、磺胺甲基异恶唑(SMZ)、磺胺嘧啶(SD),与3种金属氧化物纳米颗粒,纳米ZnO(n-ZnO)、纳米CuO(n-CuO)、纳米Cr_2O_3(n-Cr_2O_3)对发光菌(Vibriofischeri)的单一毒性及二元急性联合毒性。结果表明,SCP对V.fischeri的单一毒性明显大于其他3种磺胺;n-ZnO的单一毒性远大于n-CuO和n-Cr_2O_3;磺胺与金属氧化物纳米颗粒的二元急性联合毒性效应表现为拮抗、相加和协同,协同居多,说明金属氧化物纳米颗粒增加了磺胺的生物毒性,两者在环境中的共存带来了更大的环境生态风险。     

Leaching toxicity characteristics of municipal solid waste incineration bottom ash

The continuously increasing production of municipal solid waste incineration bottom ash (MSWIBA) has promoted its utilization as construction material and raised environmental concern. The physico-chemical properties and leaching behavior of MSWIBA were studied, and ecotoxicological testing using a luminescent bacterium bioassay was performed to assess the ecological pollution risks associated with its leached constituents. The MSWIBA was leached by two types of leachants, H₂SO₄/HNO₃ and HAc solution, at different liquid to solid ratios and contact times. The concentrations of heavy metals and anions in the leachates were analyzed. Multivariate statistical analyses, including principle component analysis, Pearson's correlation analysis and hierarchical cluster analysis, were used to evaluate the contributions of the constituents to the toxicity (EC₅₀) of the MSWIBA leachate. The statistical analyses of the ecotoxicological results showed that the Ba, Cr, Cu, Pb, F⁻ and total organic carbon (TOC) concentrations were closely correlated with the EC₅₀ value, and these substances were the main contributors to the ecotoxicity of the MSWIBA leachate. In addition, the cluster of these variables indicated similar leaching behaviors. Overall, the research demonstrated that the ecotoxicological risks resulting from MSWIBA leaching could be assessed before its utilization, which provides crucial information for the adaptation of MSWIBA as alternative materials.     

弧菌拮抗菌的筛选及其对凡纳滨对虾的抑菌防病作用

从海洋沉积物中分离到2株弧菌拮抗菌(H2和H4),经生理生化特性、16S rRNA基因序列分析分别鉴定为短小芽孢杆菌(Bacillus pumilus H2)和地衣芽孢杆菌(Bacillus licheniformis H4). H2和H4菌株均对需钠弧菌的生长有显著的抑制作用;但只有H2菌株在模拟养殖动物小肠环境中生长良好. 选择H2进行模拟试验(20尾凡纳滨对虾虾苗为1组).结果表明,对照组(不接种)虾苗存活32h,接种弧菌的试验组虾苗仅存活16h,同时接种H2菌株和弧菌的试验组虾苗存活48h,而仅接种H2菌株的试验组虾苗存活了72h,说明菌株H2可显著提高凡纳滨对虾的存活时间.现场试验中,100L养殖池中(50尾虾/池),分别接种0 (对照), 103, 104 CFU/mL的H2菌株并养殖14d,再用104 CFU/mL需钠弧菌处理14d.接种104 CFU/mL H2菌株的试验组,虾的死亡率仅为12.5%,显著低于对照组的30.8%(P0.05).     

典型城市黑臭河道水体生物毒性研究

以黑臭为特征的城市内河污染近来受到关注,检测和分析污染河流水质的生物毒性对水质标识和生态评价十分必要。本研究逐月检测了温州市九山外河(JS River)和山下河(SX River)两条典型黑臭河道水样对斑马鱼、发光细菌和热带爪蟾胚胎的毒性效应。结果显示,黑臭河道水体对斑马鱼、发光细菌(青海弧菌Vibrio Qinghaiensis Q67)和爪蟾胚胎均具有毒性效应。以斑马鱼死亡率和发光细菌相对抑光率表征的水质综合毒性较一致,且以发光细菌更为敏感。爪蟾胚胎致畸实验主要通过存活率和畸形率来表征水体的毒性。3类生物监测均显示SX水体毒性明显高于JS整体毒性,这与两条河黑臭程度相一致。通过发光细菌检测水体综合毒性的分析,显示SX和JS水体毒性在5月-8月的夏季较高,冬春季节较低,表明黑臭河道水质毒性季节性变化与水体温度(T)和溶解氧(DO)值关系密切。本研究结果可为污染水体生物毒性检测和黑臭水体综合评价及后续治理提供依据。     

城市污水厂污泥倾倒对海洋生物的毒性研究

以广州市和佛山市的5个城市污水处理厂的污泥为实验对象,选取费氏弧菌、蒙古裸腹溞、卤虫、裸项栉鰕虎鱼仔鱼4种不同营养级的海洋生物为受试对象,结合化学分析方法,研究广州区域周边污水污泥浸出液的生物毒性效应。结果显示各污水污泥浸出液的毒性均较大,且浸出液的Cu污染浓度较高;广州市污水污泥对4种受试生物的毒性效应要高于佛山市。从污泥毒性对受试生物的选择性方面分析,发现卤虫筛分能力强且灵敏度高,而费氏弧菌则相关性好且方法简捷。研究结果为反映城市污泥的生物毒性强度和选择合适毒性评价受试生物提供基础数据。     

Effectiveness evaluation of glyphosate oxidation employing the H2O2/UVC process: Toxicity assays with Vibrio fischeri and Rhinella arenarum tadpoles

The H₂O₂/UVC process was applied to the photodegradation of a commercial formulation of glyphosate in water. Two organisms (Vibrio fischeri bacteria and Rhinella arenarum tadpoles) were used to investigate the toxicity of glyphosate in samples M_1, M₂, and M₃ following different photodegradation reaction times (120, 240 and 360 min, respectively) that had differing amounts of residual H₂O₂. Subsamples of M₁, M₂, and M₃ were then used to create samples M_1,E, M_2,E and M_3,E in which the H₂O₂ had been removed. Acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) activities were measured in tadpoles to determine possible sub-lethal effects. In V. fischeri, M_1,E, which was collected early in the photodegradation process, caused 52% inhibition, while M_3,E, which was collected at the end of the photodegradation process, caused only 17% inhibition. Survival of tadpoles was 100% in samples M₂, M₃, and in M_1,E, M_2,E and M_3,E. The lowest percentages of enzymatic inhibition were observed in samples without removal of H₂O₂: 13.96% (AChE) and 16% (BChE) for M₂, and 24.12% (AChE) and 13.83% (BChE) for M₃. These results show the efficiency of the H₂O₂/UVC process in reducing the toxicity of water or wastewater polluted by commercial formulations of glyphosate. According to the ecotoxicity assays, the conditions corresponding to M₂ (11 ± 1 mg a.e. L^?1 glyphosate and 11 ± 1 mg L^?1 H₂O₂) could be used as a final point for glyphosate treatment with the H₂O₂/UV process.