转cry1Ab和epsps基因玉米C0030.3.5对大型蚤(Daphnia magna)的生态毒性研究

随着抗虫和耐草甘膦除草剂转基因玉米的迅速推广和种植,其环境安全性也越来越成为人们关注的焦点。为探讨抗虫耐除草剂转基因玉米C0030.3.5(外源基因cry1Ab和epsps)对水生动物环境的安全性,以模式生物大型蚤(Daphnia magna)为指示生物,分别使用1.5 g·L~(-1)C0030.3.5玉米粉和其非转基因对照DBN318玉米粉饲喂大型蚤28 d,探讨C0030.3.5玉米对大型蚤生长和繁殖的影响。结果显示,C0030.3.5玉米粉组大型蚤与亲本DBN318组大型蚤相比,体长、存活率、新生幼蚤总数等没有显著性差异(P0.05)。28 d饲喂实验结果表明,抗虫耐除草剂玉米C0030.3.5没有对大型蚤生长和繁殖产生不良影响。上述研究结果为转基因玉米的商业化种植和安全管理提供科学数据。     

氯霉素对大型溞的急性和慢性毒性效应研究

氯霉素是一种具有广谱杀菌作用的抗生素,曾在水产养殖中广泛使用,虽然目前已被列入我国渔药禁用清单,但在水环境中仍被大量检出。为探究氯霉素对水生生物的毒性作用,选择大型溞(Daphnia magna)作为受试生物,研究氯霉素对其急性毒性和慢性毒性效应,同时建立了氯霉素的高效液相色谱(HPLC)分析方法,通过实测浓度分析确保实验过程中氯霉素浓度保持在可接受范围内。结果表明:氯霉素对大型溞的48 h半数抑制浓度(EC50)为129.5 mg·L~(-1),95%置信区间为124.4!150.9mg·L~(-1),对溞类的急性毒性为低毒;长期暴露能抑制大型溞的产溞数量,以繁殖量为毒性指标,21 d无可观察效应浓度(NOEC)为1.25 mg·L~(-1),最低可观测效应浓度(LOEC)为2.50 mg·L~(-1);各暴露组实测浓度范围在配制浓度的80%～110%,保证了实验的有效性。同时,利用实验获得的急慢性毒性数据,计算氯霉素对大型溞的急慢性毒性比(ACR),发现利用慢性毒性求得的ACR值比利用急性毒性EC10求得的ACR值更接近推荐值。研究表明氯霉素对大型溞的急性毒性低,但具有慢性毒性效应,其环境风险不容忽视。     

农药类化合物对大型溞的毒性作用模式:与基线毒性化合物比较研究

农药在控制有害生物的同时,对水生生态系统产生较大的毒性效应。本文通过实验获得了25种基线化合物对大型溞的急性毒性数据,并与57种农药类化合物对大型溞的急性毒性数据进行比较研究,同时根据体外浓度LC_(50)、生物富集因子BCF和体内临界浓度CBR的关系,计算了这些化合物在大型溞体内的临界浓度,研究了农药类化合物对大型溞的毒性作用机理。结果表明,基线化合物在大型溞体内的临界浓度log1/CBR值在一个很窄的范围波动,而农药类化合物在大型溞体内的临界浓度log1/CBR值范围广且较高。这说明多数农药类化合物对大型溞为反应性毒性作用模式。其中,除草剂对大型溞的毒性显著低于杀菌剂和杀虫剂对大型溞的毒性。这可能是因为除草剂主要通过干扰植物光合作用、植物激素或植物分子合成发挥毒性效应,从而导致其对大型溞的生理系统难以发生反应性毒性效应。而杀虫剂和杀菌剂主要通过干扰生物神经系统、生殖系统、呼吸作用或大分子合成发挥毒性效应,因此易与大型溞生理系统发生生物化学反应,从而具有较高的毒性效应。本文分别建立了除草剂、杀菌剂和杀虫剂对大型溞的急性毒性QSAR模型。除草剂对大型溞的急性毒性机理较简单,其毒性与化合物疏水性程度和离子化程度有关;而杀菌剂对大型溞的急性毒性主要与化合物的标准生成热和极性表面积有关;杀虫剂对大型溞的急性毒性作用机理较复杂,它们对大型溞的毒性效应与其和生物分子之间的氢键和范德华力有关。     

Controlling pollution from land‐based sources in the Turkish Mediterranean coast—legislative aspects

The Turkish Mediterranean Coast is still the cleanest section in the region. This aspect needs to be strictly controlled and improved to allow for a sound development of touristic activities. Also, the adopted controlling policy should comply with the basic principles of the Mediterranean Action Plan. In Turkey, the regulations issued in connection with the “Water Products Law”; are now implemented to control wastewater discharges. Recently, major efforts were devoted to improve these regulations within the umbrella of the new “Environmental Law”;. The new proposals consider the issue from a much broader framework and provide technical support and criteria for marine outfalls, direct discharges of domestic and industrial sources and pretreatment requirement prior to discharge to the sewerage systems.     

典型化工污水处理厂废水及受纳水体的生物急性毒性监测方法应用研究

为了研究生物急性监测方法对监测典型化工污水处理厂废水的适用性,选择2家常州市典型的化工园区污水处理厂("常A"和"常B"),进行了发光细菌、藻类、大型溞和斑马鱼卵4种不同层次受试生物的急性毒性检测。研究表明,发光细菌急性毒性、藻类叶绿素荧光毒性在2个污水处理厂中均被检测到,发光细菌急性毒性通常进水大于出水,但在投放大量氧化消毒剂时,出水表现出剧毒。藻类叶绿素荧光毒性最高值出现在常B进水中。大型溞和斑马鱼卵急性毒性仅在常B进出水样中有检出,受纳河道下游水样虽未表现出急性毒性,但可观察到斑马鱼卵各类发育畸形。4种生物急性毒性检测方法中,发光细菌适用范围最广,藻类、大型溞和斑马鱼卵急性毒性方法可根据监测目的和工业园区特点相应选择。实验结论可为化工污水处理厂尾水排放过程中生物毒性监测的常态化提供方法选择,为管理部门制定排放标准提供依据。     

大型蚤对五价砷抗性选择的响应

金属污染对栖息在该环境中的生物具有强大的选择力,生物若能进化出对该金属的抗性则能在该环境中生存下去,否则将会灭绝。人工模拟的选择方法可以用来研究生物对金属的抗性进化。为了探索大型蚤(Daphnia magna)是否能进化出对五价砷(As(Ⅴ))的抗性,采用多代选择的方法对其进行了As(Ⅴ)诱导的抗性响应研究。依据本实验室大型蚤对As(Ⅴ)的96h半致死浓度(4.25 mg·L~(-1)),试验选取As(Ⅴ)亚致死浓度(8.0 mg·L~(-1))对大型蚤进行选择,每代选择30%~50%对As(Ⅴ)耐受性高的大型蚤转移至不加As(Ⅴ)环境下继续繁殖得到下一代,并重复该选择过程至获得第五代(F5)终止试验。以选择组F5代及对照组F5代大型蚤为测试目标,考察其在As(Ⅴ)继续暴露下的存活时间以期获得具As(Ⅴ)抗性大型蚤。结果显示,在F5代中,选择组大型蚤在As(Ⅴ)(8.0 mg·L~(-1))暴露下的存活时间相比对照组显著延长175%,证实了大型蚤对As(Ⅴ)的进化抗性。进一步研究发现,选择组F5代与对照组F5代相比繁殖力下降19.96%,平均每批产仔量降低15.71%。这表明经过五代人工选择后,大型蚤能够进化出对As(Ⅴ)的抗性,而这种抗性的进化伴随以生物适应性参数的降低为代价。另外,大型蚤对这种抗性的获得机制(对As(Ⅴ)的累积和脱毒机制)仍需进一步研究。     

Effect of tannins on survival and development of Daphnia menucoensis and the abundances of bacteria

Polyphenols – products of organic-matter decomposition entering water bodies from autochthonous and especially allochthonous sources – affect primary producers, bacterioplankton, and zooplankton to consequently modify food webs. Cladocerans are widely used in research experiments because they constitute the most frequent prey of high-trophic-level organisms in the majority of lakes, and certain species symbolise the ecologic prototype of the generalist filter feeder. In our study area’s shallow lakes, cladocerans, though generally of low abundance, do attain significantly high biomasses. We accordingly evaluated the mortality of the cladoceran Daphnia menucoensis Paggi, 1996 exposed to different tannin concentrations along with the bacterial abundance. In two experiments D. menucoensis females exposed to high tannin concentrations reproduced inefficiently, exhibited high mortality, and manifested altered behaviour, such as reduced reflexes and diminished mobility. Though tannins apparently affected algal abundances, the cladoceran herbivory strongly reduced those numbers. Positive effects on bacteria, however, were not recorded. High-allochthonous-organic-matter-containing water bodies support trophic webs because bacterioplankton provide an alternative energetic base for the zooplankton, as possibly occurs in shallow lakes of the Salado-River basin, characterised by cyclic hydrologic periods alternating between draught and flooding and intensive land use. In conclusion, tannins diminish D. menucoensis’s survival, locomotion, and the capacity to respond to stimuli.     

Acute and long-term effects of sucralose on the water flea Daphnia magna on mobility,survival and reproduction at different temperature regimes

Sucralose is a food additive used as a substitute for sugar. Although the risk of negative effects in organisms exposed to sucralose in the natural environment is low, it is expected that the water concentrations of sucralose will continue to increase in the near future. Temperature in the aquatic ecosystem is also expected to increase, and this needs to be considered in ecotoxicological studies. Herein, we investigated if the water flea Daphnia magna could be affected by sucralose exposure both at optimal and elevated temperatures. No increased mortality or effects on mobility and fecundity were observed after sucralose exposure at any exposure temperature during the course of the experiment. An increase in temperature to 26°С reduced the time required to produce four generations of offspring. However, this result was observed across treatments; hence speeding up of life processes was caused by an increased temperature only and not sucralose exposure. A lack of deleterious effects on the acute and chronic endpoints, even at elevated temperatures, suggests a relative robustness of this species to sucralose. However, due to the extensive use of sucralose in combination with rising water temperatures, further investigations on possible long-term effects should be studied to evaluate potential early warning signals.     

纳米二氧化铈对蛋白核小球藻和大型溞的毒性及其在大型溞体内的形态转化

随着纳米技术的飞速发展,纳米材料的应用日益广泛。同时,这类具有独特物理化学特性的微小颗粒对环境和健康的影响引起了人们的关注。本工作参考国际经济合作与发展组织(OECD)化学品生态毒理测试方法,以蛋白核小球藻(Chlorella pyrenoidosa)和大型溞(Daphnia magna)为受试生物,研究了CeO_2纳米颗粒暴露对小球藻生长、叶绿素含量和细胞内活性氧水平以及大型溞运动能力的影响,分析了大型溞体内铈的形态。随着暴露浓度的升高和时间延长,CeO_2纳米颗粒逐渐抑制小球藻的生长,导致叶绿素水平的降低和活性氧水平升高。暴露96 h后,CeO_2纳米颗粒对小球藻生长的EC50为30.4 mg·L-1,而对大型溞活动抑制的24 h、48 h-EC50分别为430.2 mg·L-1和142.7 mg·L-1。根据中华人民共和国环境保护行业标准中的毒性分级标准,CeO_2纳米颗粒对小球藻属于中毒性物质,对大型溞属于低毒性物质。CeO_2纳米颗粒在大型溞体内主要以Ce(IV)的形式存在,约有3%转化为Ce(III)。对CeO_2纳米颗粒的水生态效应给予足够重视并深入研究其毒性作用机制。     

三种双酰胺类杀虫剂制剂对大型溞生长发育和繁殖的影响

为探索双酰胺类杀虫剂对大型溞的慢性毒性,本文采用氟虫双酰胺、氯虫苯甲酰胺和溴氰虫酰胺3种双酰胺类杀虫剂制剂,测定其对大型溞生长发育和繁殖的影响,明确其对大型溞的慢性毒性以及大型溞对该类杀虫剂敏感的端点指标。结果表明,氟虫双酰胺、氯虫苯甲酰胺和溴氰虫酰胺3种制剂对大型溞的寿命、体长、蜕皮数、首胎时长、产胎数、单雌产溞数等端点指标具有不同程度的影响。3种杀虫剂显著减少大型溞蜕皮数的最低浓度分别为1.00×10-2mg·L~(-1)、1.00×10-4mg·L~(-1)、1.25×10-3mg·L~(-1);显著缩短寿命的最低浓度分别为5.00×10-3mg·L~(-1)、1.00×10~(-4)mg·L~(-1)和1.25×10-3mg·L~(-1);显著缩短体长的最低浓度分别为5.00×10-3mg·L~(-1)、8.00×10-4mg·L~(-1)和1.00×10~(-2)mg·L~(-1);显著减少产胎数的最低浓度分别为1.00×10-2mg·L~(-1)、4.00×10-4mg·L~(-1)和1.00×10-2mg·L~(-1);显著减少单雌产溞数的最低浓度分别为1.00×10~(-2)mg·L~(-1)、8.00×10-4mg·L~(-1)和5.00×10-3mg·L~(-1);但是,除溴氰虫酰胺外,其他2种杀虫剂制剂对首胎时长却没有显著影响。端点指标中对氟虫双酰胺的敏感性为寿命和体长蜕皮数、产胎数和单雌产溞数首胎时长;对氯虫苯甲酰胺为蜕皮数和寿命产胎数体长和单雌产溞数首胎时长;对溴氰虫酰胺为蜕皮数和寿命单雌产溞数体长、首胎时长和产胎数。研究结果说明,3种双酰胺类杀虫剂对大型溞的生长发育和繁殖具有不同程度的抑制作用,寿命是评价该类杀虫剂制剂对大型溞慢性毒性的最敏感端点指标。