In situ assays with tropical cladocerans to evaluate edge-of-field pesticide runoff toxicity

Tropical regions’ economy is usually based on agriculture, which involves an intensive use, and even frequent overuse, of pesticides. Nevertheless, not much research has been done on the impact of pesticides on tropical aquatic ecosystems, which are often contaminated by runoff-related pesticide inputs due to unpredictable and torrential rainfalls. This study aimed to: (i) adapt and evaluate a short-term sublethal in situ assay using post-exposure feeding as an endpoint, to the tropical cladoceran species Diaphanosoma brachyurum (collected at the Pedra do Cavalo dam in the Paraguaçu River basin, Bahia, Brazil), and, (ii) assess the role of the standard species Daphnia magna as an adequate laboratory surrogate. Lethal and sublethal (post-exposure feeding) responses were assessed for the two species. To evaluate these responses under environmentally realistic exposure conditions, a runoff event was simulated in an agricultural area previously contaminated with different deltamethrin concentrations. The resultant runoff water was used to set up microcosms with different dilutions, simulating the entrance of runoff water in an adjacent lentic system. An in situ assay with D. brachyurum was performed inside the microcosms, allowing to discriminate the effects due to deltamethrin from those due to other potential stressors associated with the experimental design (e.g. organism handling, load of suspended particles, microcosm design). Water samples were collected from microcosms to conduct a laboratory assay with D. magna. The in situ methodologies revealed to be suitable to conduct assays with D. brachyurum under tropical conditions, since all exposed organisms were successfully retrieved from the chambers. Furthermore, none of the potential stressors associated with the experimental design influenced the daphnids’ performance. The tropical cladoceran species, exposed under more realistic conditions, revealed to be more sensitive than the laboratory standard species: lethal effects were only observed for D. brachyurum and sublethal effects were noticed at a lower deltamethrin concentration for this species than for D. magna.     

Insect cell-based impedance biosensors: a novel technique to monitor the toxicity of environmental pollutants

Cell biosensors are currently emerging as novel, sensitive techniques to monitor the toxicity of environmental pollutants. Here, we have developed electric cell-substrate impedance sensing (ECIS) for on-line monitoring of the behavior of insect cells. Cells were cultured on a microarray of eight small gold electrodes, deposited on the bottom of tissue culture wells. Upon inoculation, cells showed a tendency to drift downward and attached to the gold surface precoated with the protein Concanavalin A to accelerate the cell attachment. The impedance increased because the cells acted as insulating particles to restrict the current flow. The resulting impedance, a coordination of many biological reactions within the cell, was continuously monitored in real-time to reveal information about cell spreading and micromotion. As the cell behavior was sensitive to external chemicals, the applicability of ECIS for inhibition assays was demonstrated with HgCl₂, 2,4,6-trinitrotoluene (TNT), 2-amino 4,6-dinitrotoluene (2-ADNT) and 1,3,5-trinitrobenzene (TNB). Electronic Publication     

垃圾渗滤液与再生水生物毒性的体外检测方法应用

本文总结了近年来在垃圾渗滤液和再生水生物毒性检测方面的主要体外试验模型和检测方法,并整理了这些模型和方法在生物毒性评价中的应用.目前常用的体外试验模型包括人源细胞系、其他哺乳动物细胞以及微生物细胞,相比较而言,人源细胞系在检测结果外推至人体时具有更强的说服力,因而其应用最为广泛.体外检测方法可概括为细胞毒性、遗传毒性和内分泌干扰效应检测三个方面,其中内分泌干扰效应的研究较多集中于雌激素效应.最后,本文提出开发三维体外细胞模型、体外试验与化学分析相结合、全面分析内分泌干扰效应和建立成组体外试验体系是渗滤液和再生水生物毒性检测方法的发展方向.     

通过多阶段暴露试验评价嘧菌酯对斑马鱼的急性毒性与发育毒性

嘧菌酯作为最畅销的甲氧丙烯酸酯类杀菌剂,是防治水稻病害的常用农药,其对水生生态环境的负面效应值得重视.为研究嘧菌酯对水生生物的危害,进行多阶段(成鱼、仔鱼、胚胎)斑马鱼毒性试验,分析了嘧菌酯对鱼类的急性毒性,同时,通过6 d胚胎发育试验,研究了嘧菌酯对鱼类早期发育阶段的影响.结果表明,斑马鱼3个生命阶段对嘧菌酯的敏感性(以96 h-LC50(致死中浓度)表示)顺序为:仔鱼(0.39 mg·L~(-1))胚胎(0.61 mg·L~(-1))成鱼(1.37 mg·L~(-1)).6 d胚胎发育试验结果发现,嘧菌酯可诱导斑马鱼胚胎出现一系列不良症状,包括孵化率下降、心率异常、生长抑制和心包水肿等.0.25 mg·L~(-1)的嘧菌酯可显著促进斑马鱼胚胎自主运动和心率,并能明显抑制孵化仔鱼的体长.0.6 mg·L~(-1)及更高浓度的嘧菌酯可明显抑制斑马鱼胚胎眼睛、体节、尾部和心脏的发育.研究显示,嘧菌酯对斑马鱼多个生命阶段均具有一定毒性,但对早期生命阶段毒性更强,因此,其对鱼类早期生命阶段的影响值得重视.     

Gas chromatography with atomic absorption detection for the determination of alkyl selenide compounds in the breath of mice after administration of different selenium compounds

Quantitative determination of dimethylselenide and dimethyldiselenide in the breath of mice after addition of different selenium compounds to the drinking water is described. Breath samples are collected with a cryogenic trap and analyzed by gas chromatography with atomic absorption spectrometric detection. Detection limits down to 0.2 ng/100 g body weight are obtained.

The pulmonary excretion of selenium metabolites after oral administration via drinking water or after intraperitoneal injection appears to be negligible. The bioconversion of the different selenium species is discussed.     

三种典型纳米颗粒物造成的人体肺细胞氧化应激和炎症效应(Oxidative Stress and Inflammatory Effect on A549 Cell Line Induced by Three Typical Nano-Particles )

采用体外细胞暴露实验研究了人肺腺癌细胞系(A549)单层细胞暴露于50和500μg·mL-1两种浓度纳米氧化钛、纳米氧化硅、碳纳米管和晶体石英砂等四种颗粒物后产生的氧化应激和炎症反应.用细胞活度、细胞内活性氧总量和细胞上清液中白细胞介素8(IL-8)表达量表征暴露效应.研究结果表明,纳米氧化钛、纳米氧化硅和碳纳米管在体外暴露实验过程中均发生不同程度的聚集;细胞暴露48h后,三种纳米颗粒物均使A549细胞活度下降,诱导细胞产生过量活性氧,同时刺激细胞IL-8表达量增高;三种纳米颗粒物中,纳米氧化钛和纳米氧化硅对细胞活度影响较大,碳纳米管诱发的炎症效应较另两种纳米材料强.     

Assessment of manure-application effects upon the runoff water quality by algal assays and chemical analyses

The effects of manure-application mode, rate to soil, and rainfall characteristics on the quality of agricultural runoff water have been assessed by means of the algal-growth-potential test (AGPT) and chemical analyses. This study used two modes of manure application (i.e. surface mode and incorporation mode), three manure-application rates (0, 150, 300 kg N ha(-1)), and two rainfall intensities and times (i.e. 11 mm h(-1) for 142 min and 22 mm h(-1) for 71 min). The effects of the dilution of runoff water on algal growth were also examined. The algal yields obtained with runoff from soil with the incorporated manure mode are similar to those from soil without manure application and are lower than those with the surface mode of manure application. A higher manure-application rate increases the load of nutrients in the runoff and subsequently the algal yield. The dilution of runoff water can stimulate or limit the algal growth, depending on the concentration of toxicants, N (nitrogen) and P (phosphorus) from runoff and in the aquatic diluting medium. A lower rainfall intensity plus a longer rainfall time increases algal productivity. This study showed that N is the limiting factor to algal growth at low dilution but that, at high dilution or with the incorporation mode of manure application, P becomes the limiting factor to algal growth.     

Rat cardiomyocyte H9c2(2-1)-based sulforhodamine B assay as a promising in vitro method to assess the biological component of effluent toxicity

The treatment of wastewaters is crucial to maintain the ecological status of receiving waters, and thereby guarantee the protection of aquatic life and human health. Wastewater quality evaluation is conventionally based on physicochemical parameters, but increasing attention has been paid to integrate physicochemical and biological data. Nevertheless, the regulatory use of fish in biological testing methods has been subject to various ethical and cost concerns, and in vitro cell-based assays have thus become an important topic of interest. Hence, the present study intends: (a) to evaluate the efficiency of two different sample pre-concentration techniques (lyophilisation and solid phase extraction) to assess the toxicity of municipal effluents on rat cardiomyoblast H9c2(2-1) cells, and (b) maximizing the use of the effluent sample collected, to estimate the environmental condition of the receiving environment. The gathered results demonstrate that the H9c2(2-1) sulforhodamine B-based assay is an appropriate in vitro method to assess biological effluent toxicity, and the best results were attained by lyophilising the sample as pre-treatment. Due to its response, the H9c2(2-1) cell line might be a possible alternative in vitro model for fish lethal testing to assess the toxicity of municipal effluents. The physicochemical status of the sample suggests a high potential for eutrophication, and iron exceeded the permissible level for wastewater discharge, possibly due to the addition of ferric chloride for wastewater treatment. In general, the levels of carbamazepine and sulfamethoxazole are higher than those reported for other countries, and both surpassed the aquatic protective values for long-term exposure.     

Daphne gnidium, a possible native host plant of the European grapevine moth Lobesia botrana, stimulates its oviposition. Is a host shift relevant?

Summary. We tested the hypothesis that Daphne gnidium is an ancestral host plant of Lobesia botrana. To this end, we studied the oviposition response of this moth to various aerial organs of this plant. Our results show that females prefer to lay eggs on daphne berries rather than on grapes (cv. chasselas) and that polar surface extracts of daphne berries and leaves strongly stimulate oviposition in a dosedependant way, whereas flower extracts are weakly stimulant and branch extracts are inactive or deterrent for oviposition. Furthermore, a total extract of daphne berries stimulates oviposition in semi-natural conditions when applied onto fresh grapes. Oviposition stimulants from berries were isolated by HPLC coupled with a dual-choice oviposition bioassay. The ecological value of D. gnidium for L. botrana is discussed and the possible use of oviposition stimulants from daphne, contrasting with the oviposition signal of the cultivated host plant, is proposed in the perspective of developing behavioural manipulation methods such as stimulo-deterrent diversionary strategies compatible with IPM programs.