硫丹对斑马鱼的毒性效应

硫丹是一种有机氯杀虫剂,广泛应用于谷物、蔬菜、水果、茶叶等害虫的防治.然而,在杀灭害虫、提高农业产量的同时,硫丹对水生生物的生存构成威胁.为了丰富水生生物毒理学资料,评价硫丹对水生生物健康生长的风险,本文采用传统毒理学方法,在室内静态环境条件下,研究了硫丹对斑马鱼(Dardo rerio)的毒性效应.结果显示24 h、48 h、72 h、96 h半数致死浓度(LG50)分别为4.24 μg/L、2.49 μg/L、1.77μg/L、1.62 μg/L.在亚慢性实验条件下,硫丹对肝和脑组织中的超氧化物歧化酶及乙酰胆碱酯酶有显著影响.硫丹对斑马鱼超氧化物歧化酶活性具有促进效应,在0.17～0.74μg/L范围内对乙酰胆碱酯酶具有抑制效应.  

Evidence for the induction of apoptosis by endosulfan in a human T-cell leukemic line

Several organochlorinated pesticides including DDT, PCBs and dieldrin have been reported to cause immune suppression and increase susceptibility to infection in animals. Often this manifestation is accompanied by atrophy of major lymphoid organs. It has been suggested that increased apoptotic cell death leading to altered T-B cell ratios, and loss of regulatory cells in critical numbers leads to perturbations in immune function. The major objective of our study was to define the mechanism by which endosulfan, an organochlorinated pesticide, induces human T-cell death using Jurkat, a human T-cell leukemic cell line, as an in vitro model. We exposed Jurkat cells to varying concentrations of endosulfan for 0-48 h and analyzed biochemical and molecular features characteristic of T-cell apoptosis. Endosulfan lowered cell viability and inhibited cell growth in a dose- and time-dependent manner. DAPI staining was used to enumerate apoptotic cells and we observed that endosulfan at 10-200 M induced a significant percentage of cells to undergo apoptotic cell death. At 48 h, more than 90% cells were apoptotic with 50 M of endosulfan. We confirmed these observations using both DNA fragmentation and annexin-V binding assays. It is now widely being accepted that mitochondria undergo major changes early during the apoptotic process. We examined mitochondrial transmembrane potential (_m) in endosulfan treated cells to understand the role of the mitochondria in T-cell apoptosis. Within 30 min of chemical exposure, a significant percentage of cells exhibited a decreased incorporation of DiOC₆(3), a cationic lipophilic dye into mitochondria indicating the disruption of _m. This drop in _m was both dose- and time-dependent and correlated well with other parameters of apoptosis. We also examined whether this occurred by the down regulation of bcl-2 protein expression that is likely to increase the susceptibility of Jurkat cells to endosulfan toxicity. Paradoxically, the intracellular expression of bcl-2 protein was elevated in a dose dependent manner suggesting endosulfan-induced apoptosis occurred by a non-bcl-2 pathway. Based on these data, as well as those reported elsewhere, we propose the following sequence of events to account for T-cell apoptosis induced by endosulfan: uncoupling of oxidative phosphorylation excess ROS production GSH depletion oxidative stress disruption of _m release of cytochrome C and other apoptosis related proteins to cytosol apoptosis. This study reports for the first time that endosulfan can induce apoptosis in a human T-cell leukemic cell line which may have direct relevance to loss of T cells and thymocytes in vivo. Furthermore, our data strongly support a role of mitochondrial dysfunction and oxidative stress in endosulfan toxicity.  

硫丹对小鼠红细胞免疫功能的影响

为了探讨有机氯农药硫丹对小鼠(Mus musculus)红细胞免疫功能的影响,设计了体内、外两组实验。体内实验:将40只小鼠随机分成4组,灌胃硫丹的量依次为:0、0.4、1.6、6.4mg/(kg·d)。灌胃25d后,取血测定红细胞的免疫功能。体外实验:将9只小鼠的红细胞分别与不同浓度的硫丹在体外培养,实验设空白对照组、溶剂丙酮组和4个不同浓度硫丹组,其6组实验所用硫丹的量依次为:0、0、5、10、20、40μg/ml。体外培养2h后,测定红细胞免疫粘附能力。结果表明,在活体实验中,随着硫丹浓度的增加,小鼠红细胞C3b受体花环率(ratio of C3b rosetting,C3bRR)明显下降,依硫丹灌胃浓度由低到高,其C3bRR依次为7.78%、6.80%、4.96%、4.33%;而循环免疫复合物花环率(ratio of immune complexes rosetting,ICRR)随着硫丹浓度升高而升高,分别为6.69%、6.31%、7.86%、9.42%。红细胞促NK细胞活性的功能在各组间没有显著差异。硫丹6.4mg/(kg·d)组小鼠红细胞对T淋巴细胞免疫粘附促进能力较其他3组明显降低。血浆中红细...  

Endosulfan induces oxidative stress and changes on detoxication enzymes in the aquatic macrophyte Myriophyllum quitense

Endosulfan (1) is a chlorinated insecticide still in use in both developed and emerging countries. Although its toxicity on animals has been studied in the last years, scarce information is available on its effects on plants. In this study, we exposed the aquatic macrophyte Myriophyllum quitense to environmentally relevant concentrations of endosulfan (microg/L) (1) for a short time, simulating exposures that might occur after either accidental spills or toxic run-off from agricultural areas. The main goal was to evaluate changes in both detoxication and antioxidant enzymatic systems of this plant upon exposure to endosulfan (1). Thus, we measured the activities of catalase (CAT), soluble and membrane associated glutathione-S-transferases (s- and m-GSTs) and glutathione reductase (GR), as well as the hydrogen peroxide (H2O2) content. Results showed that endosulfan (1) exerts oxidative stress on M. quitense, which was evidenced by the increase of CAT activity and the H2O2 content in exposed plants. At 5 microg/L endosulfan (1), we found a generalized induction of activities of tested enzymes, indicating that this xenobiotic activates the protection system of this plant, increasing its capacity to scavenge reactive oxygen species. On the other hand, we did not find significant changes at 0.02 microg/L endosulfan (1), which is the maximal concentration allowed for freshwater. We conclude that runoff events, which can produce significant amounts of endosulfan (1) in aquatic environments during short time, can result in oxidative stress on M. quitense, and probably on similar macrophytes.  

Optimization of environmental parameters for biodegradation of alpha and beta endosulfan in soil slurry by Pseudomonas aeruginosa

Aim: To determine optimal environmental conditions for achieving biodegradation of α‐ and β‐endosulfan in soil slurries following inoculation with an endosulfan degrading strain of Pseudomonas aeruginosa. Methods and Results: Parameters that were investigated included soil texture, soil slurry: water ratios, initial inoculum size, pH, incubation temperature, aeration, and the use of exogenous sources of organic and amino acids. The results showed that endosulfan degradation was most effectively achieved at an initial inoculum size of 600 μl (OD = 0·86), incubation temperature of 30°C, in aerated slurries at pH 8, in loam soil. Under these conditions, the bacterium removed more than 85% of spiked α‐ and β‐endosulfan (100 mg l^?1) after 16 days. Abiotic degradation in noninoculated control medium within same incubation period was about 16%. Biodegradation of endosulfan varied in different textured soils, being more rapid in course textured soil than in fine textured soil. Increasing the soil contents in the slurry above 15% resulted in less biodegradation of endosulfan. Exogenous application of organic acids (citric acid and acetic acid) and amino acids (l ‐methionine and l ‐cystein) had stimulatory and inhibitory effects, respectively, on biodegradation of endosulfan. Conclusion: The results of this study demonstrated that biodegradation of endosulfan by Ps. aeruginosa in soil sediments enhanced significantly under optimized environmental conditions. Significance and Impact of the Study: Endosulfan is a commonly used pesticide that can contaminate soil, wetlands and groundwater. Our study demonstrates that bioaugmentation of contaminated soils with an endosulfan degrading bacterium under optimized conditions provides an effective bioremediation strategy.  

Distribution of endosulfan in water, sediment and fish from Warri river, Niger delta, Nigeria

This article presents the first attempt to quantify the levels and distribution pattern of endosulfan, an organochlorine pesticide in surface water, sediment and fish ( Chrysichthys furcatus and Tilapia zilli ). The samples were collected from three stations (Ovwian, Ekakpamre and Ovu) of Warri River in the western Niger Delta of Nigeria in 2006 during the dry and wet seasons (January–August). A total of 96 samples made up of 24 samples each for water, sediment and fish were analysed in this study. The pesticide levels were analysed using high performance liquid chromatography (HPLC model CECIL 1010) to elucidate its distribution in various environmental compartments. The ranges of concentrations of the pesticide in the matrices were: 0.01–9.23 μg/l (water), 0.06–11.98 μg/gdw (sediment), 0.01–15.06 μg/gdw ( Chrysichthys furcatus) and 0.01–1.80 μg/gdw ( Tilapia zilli ). From this result, decreasing order of occurrence of the pesticide is as follows: fish > sediment > water. The concentrations observed in fish ( Chrysichthys furcatus ) were higher than the levels observed in sediment and water suggesting bioaccumulation of the pesticide by the fish. Spatial variations occurred with downstream stations having statistically higher concentrations in all matrices at P  < 0.05. Seasonal variations occurred with higher concentrations in dry season for water and sediment only, while the fish species had higher concentrations in the wet season. The observed values of endosulfan were above the ecological bench marks (0.02 μg/l) recommended by Nigeria Environmental Protection Agency and European Union. They were also relatively higher than those in previous studies on the Nigerian environment, an observation that calls for regular monitoring of the Niger Delta water bodies.  

硫丹和狄氏剂对多刺裸腹的毒性效应

为了解亚致死浓度的有机氯农药硫丹和狄氏剂对枝角类的急性、慢性和联合毒性作用,采用急性毒性实验方法研究了硫丹和狄氏剂对多刺裸腹溞(Moina macrocopa)的48 h半效应浓度,应用生命表实验方法研究了亚致死浓度的硫丹(0、52和208 μg/L)和狄氏剂(0、12和48μg/L)对多刺裸腹溞生命表统计学参数的影响.结果表明,硫丹及狄氏剂对多刺裸腹溞的48 h半效应浓度是834.9和185.8 μg/L.硫丹对多刺裸腹溞出生时的生命期望(e0)、世代时间(T)和净生殖率(R0)有显著的影响(P<0.05),但对种群内禀增长率(rm)没有显著的影响(P>0.05);狄氏剂仅对种群内禀增长率有显著的影响(P<0.05);硫丹与狄氏剂的交互作用对所有生命表统计学参数均无显著影响(P>0.05).与硫丹浓度为0时相比,52和208μg/L的硫丹均使多刺裸腹溞的e0.和T显著缩短,R0显著降低;与狄氏剂浓度为0时相比,48μg/L的狄氏剂使多刺裸腹溞的rm显著升高.这说明硫丹与狄氏剂对多刺裸腹溞的联合作用形式是独立相加,没有产生协同作用.  

Antioxidant enzyme activities and lipid peroxidation in the freshwater cladoceran Daphnia magna exposed to redox cycling compounds

Contaminant-related changes in antioxidative processes in the freshwater crustacea Daphnia magna exposed to model redox cycling contaminant were assessed. Activities of key antioxidant enzymes including catalase, superoxide dismutase, glutathione peroxidase and glutathione S-transferases and levels of lipid peroxidation measured as thiobarbituric acid-reactive substances (TBARS) and lipofucsin pigment content were determined in D. magna juveniles after being exposed to sublethal levels of menadione, paraquat, endosulfan, cadmium and copper for 48 h. Results denoted different patterns of antioxidant enzyme responses, suggesting that different toxicants may induce different antioxidant/prooxidant responses depending on their ability to produce reactive oxygen species and antioxidant enzymes to detoxify them. Low responses of antioxidant enzyme activities for menadione and endosulfan, associated with increasing levels of lipid peroxidation and enhanced levels of antioxidant enzyme activities for paraquat, seemed to prevent lipid peroxidation, whereas high levels of both antioxidant enzyme activities and lipid peroxidation were found for copper. For cadmium, low antioxidant enzyme responses coupled with negligible increases in lipid peroxidation indicated low potential for cadmium to alter the antioxidant/prooxidant status in Daphnia. Among the studied enzymes, total glutathione peroxidase, catalase and glutathione S-transferase appeared to be the most responsive biomarkers of oxidative stress.  

硫丹及其主要代谢产物对紫色土中酶活性的影响

采用室内避光培养,研究了硫丹及其主要代谢产物(硫丹硫酸盐和硫丹二醇)在紫色土中的质量浓度变化及其对土壤酶活性的影响。结果表明,α-、β-硫丹在紫色土中的消解过程符合一级反应动力学方程,半衰期分别为32—99d和69—116 d。代谢产物硫丹硫酸盐浓度在前20 d增加较快,30 d后基本趋于稳定;硫丹二醇浓度先增加后减小,15 d时达到最大。5 mg/kg的硫丹处理5 d时对脲酶和硝酸还原酶活性起激活作用,随着培养时间的增加,硫丹处理对脲酶活性从无显著影响逐渐转向抑制,第60天时抑制作用达到最大;高浓度处理(100 mg/kg)强烈抑制脲酶活性,最大降幅达到94.5%。硫丹处理(除5 mg/kg外)对硝酸还原酶活性起抑制作用,浓度越大,抑制作用越强,最大降幅达到89.9%。5 mg/kg的处理抑制多酚氧化酶活性,10—20 mg/kg的处理30 d后由激活逐渐转向抑制;100 mg/kg的处理起激活作用,第15天时酶活性达到最大,随后持续下降,第60天时恢复到对照水平。偏相关分析表明,脲酶、硝酸还原酶活性与硫丹硫酸盐浓度呈显著负相关,硫丹硫酸盐抑制这两种酶的活性;多酚氧化酶活性与α-、β-硫丹、硫丹硫酸盐和硫丹二醇浓度的偏相关性不显著。  

Klebsiella pneumoniae KE-1 degrades endosulfan without formation of the toxic metabolite,endosulfan sulfate

For bioremediation of toxic endosulfan, endosulfan degradation bacteria, which do not form toxic endosulfan sulfate, were isolated from various soil samples using endosulfan as sole carbon and energy source. Among the 40 isolated bacteria, strain KE-1, which was identified as Klebsiella pneumoniae by physiological and 16S rDNA sequence analysis, showed superior endosulfan degradation activity. Analysis of culture pH, growth, free sulfate and endosulfan and its metabolites demonstrated that KE-1 biologically degrades 8.72 microg endosulfan ml(-1) day(-1) when incubated with 93.9 microg ml(-1) endosulfan for 10 days without formation of toxic endosulfan sulfate. Our results suggest that K. pneumoniae KE-1 degraded endosulfan by a non-oxidative pathway and that strain KE-1 has potential as a biocatalyst for endosulfan bioremediation.