微囊藻毒素对褶皱臂尾轮虫的毒性效应和种群增长影响

 研究了微囊藻毒素(MC-LR)对咸淡水褶皱臂尾轮虫的急性毒性效应.采用克隆培养法,通过编制生命表计算内禀增长率(rm)、周限增长率(l)、净生殖率(R₀)和世代时间(T),研究MC-LR的浓度在1～20mg/mL之间对轮虫种群增长参数的影响.结果表明,MC-LR的24h LC₅₀为124.87mg/mL;各实验组轮虫的存活率和繁殖率及种群增长参数均低于对照组,其中以R₀的改变最明显,对照组是实验组的2.4～3.8倍;实验组轮虫的雄体发生率是对照组的2倍以上.研究还显示了轮虫对MC-LR有较强的耐受能力;MC-LR能降低轮虫的生命周期、繁殖和种群增长率;MC-LR能诱导轮虫雄性个体的发生.     

UV/H2O2工艺降解微囊藻毒素-LR

采用UV/H₂O₂联合工艺,研究了光强、初始浓度、H₂O₂投加量、pH及阴离子对微囊藻毒素-LR (MC-LR)去除率的影响.结果表明,单独H₂O₂对MC-LR基本无去除效果;单独UV工艺可以一定程度上降解MC-LR;而UV/H₂O₂联合工艺由于发生协同作用明显提高降解效率.试验发现,随着光强的增大,MC-LR的去除率不断提高;随着MC-LR初始浓度的增大,其去除率不断降低;随着H₂O₂投加量的增大,降解速率常数逐渐增大,当H₂O₂投加量由1 mmol/L增大到3 mmol/L时,降解速率常数由0.084 4上升到0.166 4;当pH为3.13时,在相同条件下MC-LR的去除效果最好;阴离子的投加不利于MC-LR的降解,其中CO^2-₃、NO^-₃影响最大.     

Role of nitric oxide in the genotoxic response to chronic microcystin-LR exposure in human-hamster hybrid cells

Microcystin-LR (MC-LR) is the most abundant and toxic microcystin congener and has been classified as a potential human carcinogen (Group 2B) by the International Agency for Research on Cancer. However, the mechanisms underlying the genotoxic effects of MC-LR during chronic exposure are still poorly understood. In the present study, human-hamster hybrid (A_L) cells were exposed to MC-LR for varying lengths of time to investigate the role of nitrogen radicals in MC-LR-induced genotoxicity. The mutagenic potential at the CD59 locus was more than 2-fold higher (p < 0.01) in A_L cells exposed to a cytotoxic concentration (1 μmol/L) of MC-LR for 30 days than in untreated control cells, which was consistent with the formation of micronucleus. MC-LR caused a dose-dependent increase in nitric oxide (NO) production in treated cells. Moreover, this was blocked by concurrent treatment with the NO synthase inhibitor N^G-methyl-_L-arginine (L-NMMA), which suppressed MC-LRinduced mutations as well. The survival of mitochondrial DNA-depleted (ρ⁰) A_L cells was markedly decreased by MC-LR treatment compared to that in A_L cells, while the CD59 mutant fraction was unaltered. These results provided clear evidence that the genotoxicity associated with chronic MC-LR exposure in mammalian cells was mediated by NO and might be considered as a basis for the development of therapeutics that prevent carcinogenesis.     

检测2,4-D的一次性安培型免疫传感器

2,4-D是广泛使用的除草剂,以2,4-D为检测目标,研究了安培型免疫传感器.首先采用1-乙基-3-(3-二甲基氨丙基)碳二亚胺盐酸盐(EDC)作为交联剂制备2,4-D与BSA以及PLL的偶联物.用结合比1∶16的2,4-D与偶联物制备兔抗血清.免疫传感器的制备包括2个步骤,使用丝网印刷工艺制备一次性碳电极,然后通过戊二醛交联在电极表面固定2,4-D与多聚赖氨酸(PLL)的偶联物.待测水样与2,4-D抗血清和HRP酶标二抗进行间接免疫竞争反应.在HRP酶在邻苯二胺和H₂O₂的共同作用下发生酶促反应,检测还原电流以反映2,4-D的浓度.试验了不同浓度抗血清和酶标二抗对检测信号的影响,结果表明,2,4-D的检测下限达到1.69ng/mL,线性区间1.69～30 000ng/mL,适合饮用水中2,4-D的检测要求.     

Ru负载促进GeZnNO固溶体光解水反应的机理研究

光催化水分解生成氢气和氧气是解决环境污染和能源危机的方法之一,但当前对光解水的反应机理研究较少.本文通过进一步研究反应路径,以期为合成高效率光解水反应光催化剂提供新思路.采用固态合成法制备了GeZnNO可见光催化剂,并对部分催化剂进行RuO₂改性.同时,通过XRD、BET、UV-vis-DRS、PL等方法对催化剂进行表征.在整个光催化水分解体系中加入可能的中间产物H₂O₂,分别观察?OH和O₂的变化,初步证明H₂O₂是水分解中间产物,并提出相应水分解反应机理.通过在整个光催化水分解体系中加入Fe³⁺,观察?OH和O₂的变化, 进一步验证了前面的推测.最终结果表明,H₂O₂是光催化水分解反应体系中氧气生成的中间产物,对整个水分解起着重要作用.同时发现,RuO₂助催化剂不仅具有析氢位点的作用,还能够催化H₂O₂分解产生O₂,促进水分解的进行.该研究结果可为光催化水分解性能的进一步提高提供理论参考.     

氧化剂增强TiO2纤维光催化降解DMF研究

研究TiO₂纤维光催化降解DMF水溶液的结果表明:氧化剂增强了TiO₂纤维对DMF的降解能力,在其它反应条件相同情况下,O₃/TiO₂(F)对DMF降解率是air/TiO₂(F)和H₂O₂/TiO₂(F)的1.5倍左右,且降解速率提高2倍左右,COD分析表明DMF几乎完全矿化.同时实验过程中确定了DMF光催化降解中间产物仲胺的存在.以二甲胺为仲胺代表物进行光催化降解研究表明,中间产物仲胺的迅速降解是O₃增强TiO₂纤维降解效应的主要因素.     

常压和真空下CS2的光氧化反应

利用原位FTIR、XPS 和GC-MS等技术手段,研究了常温紫外光辐照下常压及真空状态中,CS₂的光氧化反应.结果表明,常压及真空状态下CS₂光氧化反应的产物均为COS、SO₂、CO 和 CO₂,反应均遵循表观一级反应动力学规律,但中间产物COS的浓度变化有较大的差异.常压下COS浓度先迅速增大,然后减少; 真空条件下COS浓度增加到常压下COS浓度的 1/10后基本持平,然后逐渐减小,整个变化趋势缓慢; 2种条件下CS₂的半衰期分别为31.6 s和39.8 s, 中间产物COS降解 98% 所需时间分别为 55 min和22 min.在相同的光照条件下,O₂浓度的增加有助于CS₂光氧化反应的进行.以CS₂光氧化中间产物COS为初始物的光氧化反应主要产物为CO₂和SO₂,后者可以进一步被氧化为SO2-4.适量增加O₂的浓度可以促进COS光氧化反应的进行,但O₂的浓度增加至一定程度后反应速率基本保持恒定.以上结论对揭示CS₂和COS的源和汇以及全球S物质循环具有重要价值.     

电化学方法降解酸性红B研究

研究了成对电氧化技术降解酸性红B染料废水过程.通过高效液相色谱(HPLC)对阴阳极室降解过程中的产物进行了检测,结合UV-Vis变化曲线及TOC、COD_Cr变化过程,发现阳极室脱色比较快,生成多种中间物质;阴极室中间产物很少且降解更完全.电解140min时阴极室中TOC和COD_Cr去除率分别达到71.70%和56.40%,而阳极室中去除率分别为25.15%和27.57%.实验结果表明,阴阳极室中生成氧化物质的氧化能力不同而使酸性红B在阴阳极室按不同的机理降解.     

产乙烯脱卤拟球菌195脱卤反应途径的理论探针

运用密度泛函理论在B3LYP/6-31G(d)水平下对多氯二苯并-对-二 (PCDDs)、多氯二苯并呋喃(PCDFs)、多氯联苯(PCBs)、多溴联苯醚(PBDEs)和氯苯分子进行计算,找到了一个可指示产乙烯脱卤拟球菌(Dehalococcoides ethenogenes)195降解转化这些芳香卤化物脱卤反应途径和中间产物的参数—结构上可能的脱卤产物的分子总能量(ET).以ET作为理论探针可指示PCDD/Fs、PCBs、PBDEs和氯苯被菌株195转化的主要脱卤中间产物,脱卤反应倾向于生成具有较低ET值的中间产物.另外可以利用结构上可能的脱卤中间产物与具有最低ET值脱卤中间产物间的分子总能量差(?ET)作为理论探针,推断存在第2种脱卤中间产物的可能性,?ET越小,则同时存在多种脱卤中间产物的可能性越大.     

利用SPR技术测定湖水中微囊藻毒素

利用表面等离子共振(SPR)技术,建立了快速定量检测湖水中微囊藻毒素LR(MC-LR)的方法.将MC-LR共价偶联到表面等离子共振芯片表面,检测了芯片的稳定性,并利用免疫竞争抑制原理构建标准曲线,对湖水样品进行了检测.结果表明:制备的芯片信号稳定,50个循环相对标准偏差(RSD)为0.62%;该方法的最低检测浓度为0.25ng/mL,可作为水体中MC-LR的分析检测方法.     

Role of nitric oxide in the genotoxic response to chronic microcystin-LR exposure in human–hamster hybrid cells

Microcystin-LR (MC-LR) is the most abundant and toxic microcystin congener and has been classified as a potential human carcinogen (Group 2B) by the International Agency for Research on Cancer. However, the mechanisms underlying the genotoxic effects of MC-LR during chronic exposure are still poorly understood. In the present study, human–hamster hybrid (A_L) cells were exposed to MC-LR for varying lengths of time to investigate the role of nitrogen radicals in MC-LR-induced genotoxicity. The mutagenic potential at the CD59 locus was more than 2-fold higher (p < 0.01) in A_L cells exposed to a cytotoxic concentration (1 μmol/L) of MC-LR for 30 days than in untreated control cells, which was consistent with the formation of micronucleus. MC-LR caused a dose-dependent increase in nitric oxide (NO) production in treated cells. Moreover, this was blocked by concurrent treatment with the NO synthase inhibitor N^G-methyl-l-arginine (l-NMMA), which suppressed MC-LR-induced mutations as well. The survival of mitochondrial DNA-depleted (ρ⁰) A_L cells was markedly decreased by MC-LR treatment compared to that in A_L cells, while the CD59 mutant fraction was unaltered. These results provided clear evidence that the genotoxicity associated with chronic MC-LR exposure in mammalian cells was mediated by NO and might be considered as a basis for the development of therapeutics that prevent carcinogenesis.     

Microcystin-LR biodegradation by Sphingopyxis sp. USTB-05

A promising bacterial strain for biodegrading microcystin-LR (MC-LR) as the sole carbon and nitrogen source was successfully isolated from Lake Dianchi, China. The strain was identified as Sphingopyxis sp. USTB-05, which was the first isolated MCs-biodegrading Sphingopyxis sp. in China. The average biodegradation rate of MC-LR by Sphingopyxis sp. USTB-05 was 28.8 mg·L⁻¹ per day, which was apparently higher than those of other bacteria reported so far. The optimal temperature and pH for both strain USTB-05 growth and MC-LR biodegradation were 30°C and 7.0, respectively. The release of MC-LR from the cyanobacterial cells collected from Lake Guishui and the biodegradation of MC-LR by both strain and cell-free extract (CE) were investigated. The results indicated that MC-LR with the initial concentration of 4.0 mg·L⁻¹ in water was biodegraded by Sphingopyxis sp. USTB-05 within 4 d, while MC-LR with the initial concentration of 28.8 mg·L⁻¹ could be completely removed in 3 h by CE of Sphingopyxis sp. USTB-05 containing 350 mg·L⁻¹ protein. During enzymatic biodegradation of MC-LR, two intermediate metabolites and a dead-end product were observed on an HPLC chromatogram. Moreover, the similar scanning profiles of MC-LR and its metabolic products indicate that the Adda side-chain of MC-LR was kept intact in all products.     

微囊藻毒素-LR对恶臭假单胞菌细胞活性和表面特性的影响

在恶臭假单胞菌(Pseudomonas putida)降解微囊藻毒素-LR(MC-LR)的体系中,研究了菌体细胞表面特性和活性的变化,考察了MC-LR对菌体的影响,对MC-LR的微生物毒性作用进行了初步的探索.结果表明,MC-LR可以使菌体细胞质膜通透性增加,并影响菌体离子代谢及可溶性蛋白等物质的分泌.当MC-LR质量浓度由0增大到2.0 mg·L-1时,能促进菌体分泌和释放可溶性糖及Na+、Cl-等离子.流式细胞实验检测发现,MC-LR加速了菌体细胞的死亡,且死亡率随着MC-LR质量浓度的增加而增大,2.5 mg·L-1MC-LR作用5 d,比未添加MC-LR的对照体系增加了近30%.扫描电镜观察表明,在MC-LR作用下,菌体细胞的微观形貌发生了改变,细胞褶皱加深,在2.5 mg·L-1MC-LR存在下培养5 d后,大部分细胞破裂,细胞内含物流出,细胞结构损坏严重.     

Characterization of chlorine dioxide as disinfectant for the removal of low concentration microcystins

Microcystins, which represents one kind of cancerogenic organic compounds, is abundant in eutrophication water. The effects of reaction factors on chlorine dioxide (ClO₂) for removal of low-concentration Microcystin-LR, Microcystin-RR, and Microcystin-YR in water as well as the reaction mechanisms was investigated by using enzyme-linked immunosorbent assay (ELISA) kit and gas chromatography-mass spectrometry (GC-MS). The results showed that MC-LR, MC-RR, and MC-YR could be efficiently decomposed by ClO₂. The degradation efficiency was shown positively correlated to the concentration of ClO₂ and reaction time; while the effect of reaction temperature and pH is slight. The kinetic constants and activation energies of the reaction of MC-LR, MC-RR, and MC-YR with ClO₂ are determined as 459.89, 583.15, 488.43 L·(mol·min)⁻¹ and 64.78, 53.01, 59.15 kJ·mol⁻¹, respectively. As indicated by high performance liquid chromatography mass spectrometer (HPLC-MS) analysis, degradation should be accomplished via destruction of Adda group by oxidation, with the formation of dihydroxy substituendums as end products. This study has provided a fundamental demonstration of ClO₂ serving as oxidizing disinfectant to eliminate microcystins from raw water source.     

筛选菌种酶催化降解微囊藻毒素的特点

从云南滇池底泥中筛选出1株能够高效降解微囊藻毒素(Microcystins,MCs)的纯菌种.该菌无细胞提取液能进行降解MC-RR和MC-LR的酶促反应,并有最终产物的积累.在pH 6～8范围内酶对MC-RR和MC-LR的催化降解活性较高,Cu²⁺、Mn²⁺和Zn²⁺对降解MCs的酶促反应没有明显的促进作用.     

微囊藻毒素完全抗原的研制

为研制通用性微囊藻毒素（Microcystins,MCs）完全抗原,首先选用硫醇盐将微囊藻毒素氨基衍生化,利用戊二醛两步法合成完全抗原,戊二醛的两个醛基分别与微囊藻毒素和牛血清白蛋白上的氨基发生反应,形成稳定的Schiff碱,从而达到微囊藻毒素的完全抗原化。结果显示,偶联比在3～18：1,平均偶联比为8：1,可作为下一步免疫的完全抗原使用。     

环境样品免疫检测基质效应分析与控制

免疫检测技术以其特异性强、灵敏度高、分析容量大、反应速度快、检测费用低廉等优点,逐渐成为环境样品大规模快速筛查和预警监测的首选技术.由于免疫检测无需样品预处理、能实现对样品的在线检测,因此环境样品中复杂的基质可能对免疫检测产生非常大的干扰和影响.以水环境中微囊藻毒素-LR的酶联免疫吸附试验（ELISA）检测为基础,系统地考察了环境样品中各种背景基质对ELISA检测的影响,评价了各个基质的影响规律和程度,并研究提出了相应的控制和消除各不利影响的方法,最后统一提出水环境样品免疫检测抗基质效应干扰的控制方案,即采用10×PBS＋1%BSA＋0.5%EDTA作为免疫检测反应的缓冲体系,能有效消除各基质效应对免疫检测的干扰.该方案同样适应于其它不需要对环境样品进行预处理的免疫检测技术.     

基于太湖微囊藻毒素的叶绿素a阈值研究

以探讨叶绿素a(Chl-a)阈值为主要目的于2013年6~10月间每月在太湖采样,通过固相萃取超高效液相色谱串联质谱法对水体中3种微囊藻毒素(MC-LR、MC-RR、MC-YR)进行检测.利用SPSS软件分析各种形态的微囊藻毒素(MCs)与总氮(TN)、总磷(TP)、Chl-a、高锰酸盐指数等富营养指标的相关性,分析了MC-LR、MCs与Chl-a的关系.结果表明,太湖MCs污染较严重,其浓度的空间分布特征为:梅梁湾>贡湖、西部沿岸区>湖心区>胥湖区、南部湖区,并以MC-LR浓度最高;相关性分析表明MC-LR、MC-RR、MC-YR及MCs均与高锰酸盐指数、TN、TP、Chl-a呈极显著正相关(P<0.01).结合饮用水中MC-LR和MCs的标准限值分析得出,太湖Chl-a的阈值是12.26 mg·m-3,与美国北卡罗莱纳州湖泊的Chl-a标准值比较属于安全阈值,具有一定的科学性.     

微囊藻毒素-LR多克隆抗体的制备

通过对新西兰大白兔免疫自制的微囊藻毒素-LR(Microcystin-LR,MC-LR)完全抗原BSA-MC-LR,获得了质量较好的抗MC-LR的多克隆抗体,间接ELISA表明抗体的效价能达到1.5×10⁵;固定包被抗原OVA-MC-LR,采用间接竞争ELISA测定水体中的微囊藻毒素,标准曲线表明对水样中MC-LR的检测下限为10ng/L,线性区间为30ng/L·3μg/L,能满足对饮用水和地表水中MC-LR的检测要求.