Real-time fluorescent quantitative immuno-PCR method for determination of fluoranthene in water samples with a molecular beacon

A reliable and sensitive competitive real-time fluorescent quantitative immuno-PCR (RTFQ-IPCR) assay using a molecular beacon was developed for the determination of trace fluoranthene (FL) in the environment. Under optimized assay conditions, FL can be determined in the concentration range from 1 fg/mL to 100 ng/mL, with y = 0.194x + 7.859, and a correlation coe cient of 0.967 was identified, with a detection limit of 0.6 fg/mL. Environmental water samples were successfully analyzed, recovery was between 90% and 116%, with intra-day relative standard deviation (RSD) of 6.7%–12.8% and inter-day RSD of 8.4%–15.2%. The results obtained from RTFQ-IPCR were confirmed by ELISA, showing good accuracy and suitability to analyze FL in field samples. As a highly sensitive method, the molecular beacon-based RTFQ-IPCR is acceptable and promising for providing reliable test results to make environmental decisions.     

Toxicity of carbaryl,diquat dibromide,and fluoranthene,individually and in mixture,to larval grass shrimp,Palaemonetes pugio

This study examined the toxicity of two pesticides (carbaryl and diquat dibromide) and one polycyclic aromatic hydrocarbon (fluoranthene), both singly and in mixture, to grass shrimp larvae (Palaemonetes pugio). These three chemicals are all present in coastal environments and can easily enter estuarine ecosystems. Fluoranthene was the most toxic chemical with a 96-h LC₅₀ value of 32.45 μ g/L, followed by carbaryl (43.02 μ g/L) and diquat dibromide (1624 μ g/L). In the chemical mixture tests, the binary carbaryl/diquat dibromide mixture and the ternary carbaryl/diquat dibromide/fluoranthene mixture had additive results.     

十六烷基三甲基氯化铵与荧蒽对小球藻的联合毒性及其作用机制

以普通小球藻(Chlorella vulgaris)为受试生物,采用批量培养方法研究了阳离子表面活性剂十六烷基三甲基氯化铵(CTAC)与多环芳烃荧蒽(Flu)的联合毒性及其作用机制.结果表明,当CTAC初始浓度固定为100μg/L时,随Flu浓度的升高(0~100.84μg/L), CTAC与Flu的联合毒性由协同效应(0~22.50μg/L)转为拮抗效应(22.50~100.84μg/L).当Flu浓度为1.13μg/L时,协同效应达到最大(RI＝2.01),与对照组相比,生物量抑制率从37.5%增加至80.9%;对氮和铁单位吸收量分别从0.27mg和9.18μg降至0.09mg和2.14μg;藻细胞Zeta电位从-10.0mV提高至-8.3mV;叶绿素a和可溶性蛋白质含量分别从5.00mg/L和80.65μg/mg降为2.57mg/L和50.36μg/mg.根据实验结果分析, CTAC与Flu复合污染体系提高了藻细胞Zeta电位,抑制了小球藻对氮和铁的吸收,降低了藻细胞体内叶绿素和蛋白质的含量.     

平邑甜茶根毛细胞离子流动性对PAHs胁迫的响应

以苹果砧木-平邑甜茶（Malus hupehensis Rehd）幼苗为实验材料,采用营养液栽培,利用非损伤微测技术,研究多环芳烃（PAHs）中荧蒽和苯并（b）荧蒽胁迫处理对平邑甜茶幼苗根毛细胞Ca²⁺、K⁺、H⁺流速的影响.结果表明：（1）经荧蒽及苯并（b）荧蒽胁迫处理后,平邑甜茶幼苗根毛细胞Ca²⁺平均流速由对照的（-63.53±9.30）pmol/（cm²×s）分别增加到（+62.85±10.00）pmol/（cm²×s）、（91.33±19.72）pmol/（cm²×s）;K⁺流速平均值由基础流速（-60.56±14.56）pmol/（cm²×s）分别增至（+32.60±5.44）pmol/（cm²×s）、（+36.76±5.23）pmol/（cm²×s）;H⁺平均流速由对照的（+44.38±5.19）pmol/（cm²×s）分别降低至（-0.72±0.055）pmol/（cm²×s）、（-6.34±0.79）pmol/（cm²×s）.经荧蒽及苯并（b）荧蒽处理,根毛细胞表面Ca²⁺、K⁺、H⁺流动性发生明显逆转.Ca²⁺、K⁺表现为外排趋势,且外排量逐渐降低,最终趋于稳定;H⁺表现较稳定的内吸趋势.（2）苯并（b）荧蒽胁迫对平邑甜茶幼苗根毛细胞离子流动性造成的毒性效应高于荧蒽.说明PAHs胁迫会破坏植物根毛细胞离子流动性,影响植物正常生长,为深入研究植物受PAHs胁迫所产生的响应提供理论依据.     

一株荧蒽降解菌的筛选鉴定及降解特性

采用富集培养方法,从多环芳烃污染的土壤中分离筛选到一株能以高分子量的多环芳烃荧蒽为惟一碳源和能源且生长状况良好的菌株DN002。通过形态观察、生理生化指标及16S rRNA同源序列分析比对,结果表明,该菌株为木糖氧化产碱菌(Achromobacter xylosoxidans),最适生长温度为32℃,最适生长pH为7～7.5。该菌株对荧蒽有较强的降解能力,在14 d内对500 mg/L的荧蒽的降解率为92.8%。菌株细胞蛋白SDS-PAGE结果显示,经荧蒽诱导后,在分子量为18～66 kDa范围内有显著的差异蛋白条带。     

粉煤灰沸石负载Ce^3＋／TiO2光催化降解水中的菲和荧蒽

以钛酸四丁酯为前驱体，粉煤灰合成沸石为载体，采用溶胶-凝胶方法，在低温条件下制备了稀土铈掺杂的TiO2光催化剂。利用SEM—EDS、XRD、FTIR对催化剂进行了分析和表征。以高压汞灯为灯源，对多环芳烃菲、荧蒽的降解进行了研究。实验考查了稀土铈掺杂质量分数、催化剂用量、溶液pH、目标物初始质量浓度等因素对光催化降解的影响，研究了其光降解动力学。结果表明，当稀土铈含量为0．5％，催化剂用量为3g／L，pH偏碱性时，催化效果最佳。光催化反应符合Langmuir—Hinshelwood动力学规律，菲、荧蒽的降解过程符合一级反应动力学，反应速率常数分别为0．0126min^-1，0．0099min^-1。     

Studies in the Biodegradation of 5 PAHs (Phenanthrene,Pyrene, Fluoranthene,Chrysene und Benzo(a)pyrene) in the Presence of Rooted Poplar Cuttings

Cuttings of Populus nigra L. cv. Loenen were cultivated in sand treated with one of the following PAHs: phenanthrene (Phen), fluoranthene (Flt), pyrene (Pyr), chrysene (Chr) and benzo[a]pyrene (BaP). The PAHs were applied at varying levels of concentration to each test series. After 6 weeks the concentration and the distribution of the PAHs in the substrate of the various sets of tests were compared with the concentration in the substrate of the control. Additionally the substrate and the plant roots were tested for evidence of degradation products of PAHs. The results revealed that the levels of concentration of Phen and Pyr detected in the substrate surrounding the roots was in some cases significantly lower than in the corresponding section of substrate in the unplanted set (= control). This phenomenon did not occur for Flt and BaP and in the case of Chr only in those substrates, which had been treated with the highest levels of concentration. As the presence of lesser amounts of Phen and Pyr in the plant pots cannot only be attributed to their accumulation and metabolism in the roots, it is fair to assume that the chemical transformation of these three PAHs took place outside the roots. The set of tests treated with Phen revealed the presence of 2- or 3-hydroxy-Phen (main components), a hydroxy-methoxy-Phen, 9,10-Phenanthrenequinone and one unidentified compound in metabolite form. Altogether eleven metabolites of Pyr were identified in the root extracts, which can be divided into three groups: 1-Hydroxy-Pyr and derivatives, dihydroxy-Pyr and derivatives and ring fission products (4-Hydroxy-Pyr and a derivative of the 4-Phen-carbonic acid). However, the metabolite mass detected for Phen and Pyr represents only an insignificant percentage in comparison with the lesser amounts of PAHs observed in the planted set of tests. This indicates that the three PAHs were reduced to lower molecular compounds, which are methodically impossible to record, and subsequently translocated to other parts of the plant and integrated into the biomass. Although no lesser amount for Flt and BaP was found in the plant pots, 1-Hydroxy-Flt, an unidentified compound of Flt and 1-Methoxy-BaP were detected. These are presumably end products which were enhanced in the roots. It was not possible to identify any transformation products of Chr. It can be assumed that the majority of metabolites were not synthesised in the roots but are a result of microbial degradation in the rhizosphere. The test plants improved the conditions for the biotransformation of Phen and Pyr significantly and accumulated Flt, Pyr, Chr and BaP in their roots. It can therefore be concluded that the use of plants in the bioremediation of contaminated soils is a promising option.     

荧蒽对大豆种子发芽的影响

针对国内外有关荧蒽(FLT)对种子萌发影响研究极少的现状,本文设置5个质量浓度(2,5,15,25,50 mg·L-1)的染毒组、1个空白对照组(0 mg·L-1)和1个助溶剂对照组(丙酮体积分数为1%),以大豆种子发芽率、发芽势、发芽指数、活力指数和幼苗长度为测量指标,探索性地研究了FLT对大豆种子萌发的毒性效应。结果表明:在浓度小于15 mg·L-1的FLT胁迫下,大豆发芽率与对照相比无显著差异,大于等于15 mg·L-1的FLT胁迫则显著抑制了发芽率(p0.05)。发芽势、发芽指数、活力指数则对FLT的胁迫作用更加敏感,当FLT浓度大于等于5 mg·L-1时呈显著降低(p0.05)。FLT对幼苗长度起抑制作用,且随浓度越高和处理时间增长,抑制作用逐渐增强。     

生物扰动作用对河口沉积物中荧蒽去除的影响

通过室内微宇宙实验系统研究了天津厚蟹(Helice tientsinensis)和双齿围沙蚕(Perinereis aibuhitensis)生物扰动作用下河口沉积物中荧蒽的去除情况。实验结果显示,天津厚蟹扰动组中荧蒽的去除率显著高于沙蚕扰动组(P=0.05)和对照组(P=0.003),其中对表层(0~2 cm)和中层(3~5 cm)的促进效果最为显著;虽然各实验组表层沉积物中荧蒽的去除率均超过50%,但扰动组的去除更快,在36 d时就达最高去除率68%;双齿围沙蚕扰动组底层沉积物中荧蒽的去除率高于厚蟹扰动组和对照组,但差异不显著。研究表明表层沉积物中的荧蒽易去除,厚蟹生物扰动对荧蒽去除有显著促进作用;在距离表层5cm以下的沉积物中荧蒽的持久性增强,但生物扰动作用可促进其去除。     

Effects of benzo(k)fluoranthene,a polycyclic aromatic hydrocarbon on DNA damage,lipid peroxidation and oxidative stress in marine gastropod Morula granulata

Benzo(k)fluoranthene [B(k)F] is one of the widespread priority pollutants of polycyclic aromatic hydrocarbons that has been scarcely studied for exposure assessment. With studies reporting a high amount B(k)F in sediments and water samples around the world, it has become vital to study its effects on aquatic organisms. In this connection, this study is conducted to study the effect of different concentrations of B(k)F (1, 10, 25 and 50?µg/L) in marine gastropod Morula granulata exposed in vivo for 96?h. A concentration-dependent increase in percentage tail DNA (TDNA) as measured by comet assay was observed in snails exposed to B(k)F. Exposure concentrations above 1?µg/L B(k)F showed significant increase in superoxide dismutase (SOD) activity and lipid peroxidation value in snails. After 96?h, SOD activity was found to be doubled for 50?µg/L B(k)F in comparison to control. A significant increase in catalase and glutathione S-transferase activity was observed at all exposure conditions at the end of the exposure time. Our study showed that B(k)F induces oxidative stress in snails which further lead to genotoxic damage. To our knowledge, this is the first study on oxidative stress and genotoxic damage in gastropods exposed to B(k)F.