Rhodobacter sp. NP25b菌株缺氧降解壬基酚聚氧乙烯醚的研究

从城市污水处理厂活性污泥中分离得到一株能够在缺氧条件下以壬基酚聚氧乙烯醚(NPEOs)为惟一碳源和能源生长的菌株NP25b.经生理生化鉴定和16S rRNA基因序列分析,该菌株属于红细菌属(Rhodobacter sp.),对该菌株降解NPEOs的特性进行了研究.结果表明,在缺氧条件下,菌株NP25b在7 d内对初始底物浓度为400 mg/L NPEOs的降解率可达84%.利用液相色谱-质谱(LC-MS)和气相色谱-质谱(GC-MS)对NPEOs降解中间产物进行了分析,结果表明,主要降解产物为短链NPEOs和壬基酚聚氧乙烯醚乙酸(NPECs),其中包括具有较强内分泌干扰效应的NP1EO.该菌株能够代谢含有疏水基团的聚氧乙烯醚类表面活性剂,例如辛基酚聚氧乙烯醚和脂肪醇聚氧乙烯醚.推测菌株NP25b降解NPEOs是通过乙氧基(EO)链末端氧化后逐步切割完成的.     

水溶液中活性成分对壬基酚聚氧乙烯醚光降解的影响

采用长波紫外灯(UV365)和模拟日光灯研究光化学活性成分对水溶液中壬基酚聚氧乙烯醚(NPEOs)光降解效率的影响,并讨论了光降解导致NPEOs成分分布的变化.研究表明,在UV365辐射下,亚硝酸根,Fe(Ⅲ),H2O2和腐殖酸(HA)等的存在对NPEOs96h的降解效率有较明显的促进作用;在模拟日光下,Mn(Ⅱ),Fe(Ⅲ),H2O2等对NPEOs的光降解也有一定的促进,而此条件下HA的存在则表现为对光降解的抑制作用.此外,还考察了NPEOs在不同比例的Fe(Ⅲ)-HA和Fe(Ⅲ)-H2O2复合体系中的光降解效率.除了导致光降解效率的提高外,活性成分的存在也会导致降解后NPEOs成分分布的变化.以在UV365辐射下为例,H2O2(1000 μmol·l-1)或Fe(Ⅲ)(10和100 μmol·l-1)的加入使反应后短链NPEOs所占比例从初始的22.52%明显提高至29.75%,29.89%和51.89%.这种成分分布的变化可能是由于长链NPEOs在降解过程中发生EO链衰减,从而使短链NPEOs成为其降解中间产物;或是由于长链NPEOs比其短链同系物具有更高的敏化降解速率所导致的.     

Growth factors, kinetics and biodegradation mechanism associated with Pseudomonas nitroreducens TX1 grown on octylphenol polyethoxylates

The growth properties and biodegradation mechanism of a Gram-negative bacterium, Pseudomonas nitroreducens TX1 that was able to grow on branched octylphenol polyethoxylates (OPEO(n), average n=9.5) as the sole carbon source over a wide concentration range (1-100,000 mgl(-1)) were studied. Analysis of growth factors indicated the highest specific growth rate (micro) of 0.53 h(-1) was obtained at an initial concentration of 5,000 mgl(-1) OPEO(n). An optimal C/N ratio of 12 was obtained for (NH(4))(2)SO(4) as the nitrogen source in a cultivated medium at pH 7. The kinetic analysis demonstrated that bacterial growth and OPEO(n) degradation followed the Monod equation and were based on a substrate concentration inhibition model and pseudo-first-order reaction, respectively. The substrate inhibition coefficient was over 18,000 mgl(-1) and this indicates that the strain has an ability to sustain growth at high concentrations of OPEO(n) and use it as the sole carbon source under such a stress condition. Furthermore, LC-MS analysis showed that the biodegradation mechanism of dodecyl octaethoxylate (AEO8) by P. nitroreducens TX1 was the sequential cleavage of the ethoxylate chain.     

光电催化降解壬基酚影响因素研究

采用sol-gel法制备TiO2薄膜光电极,以该电极为工作电极,铂丝作对电极,饱和甘汞电极为参比电极,对壬基酚的光电催化降解进行了研究。结果表明:该电极具有n型半导体的特征行为。在外加偏压为+0.8V、pH=5、、30mg/L的H2O2中对初始浓度为20mg/L的壬基酚溶液光照180min降解率达62.22%。讨论了氧的存在、外加偏压、pH等因素对光电催化反应的影响。     

两株壬基酚和双酚A高效降解菌株的筛选和降解特性

以桂林市上窑污水处理厂污泥脱水车间剩余污泥、上窑堆肥厂的堆肥堆料和桂林雁山镇森林土壤为菌源进行驯化,分离纯化并筛选得到2株能分别以壬基酚(NP)和双酚A(BPA)为唯一碳源和能源生长的降解菌株N-1和B-1。通过对菌株的16S r DNA序列同源性分析,初步鉴定N-1和B-1菌分别为Cupriavidus(贪铜菌属)、Acinetobacter(不动杆菌属)。通过两菌株分别降解NP和BPA的单因素实验,确定了降解动力学以及时间、温度、p H值对降解过程的影响。研究结果表明,细菌N-1,B-1的最佳初始目标污染物质量浓度为5~10 mg/L,降解40 h,N-1去除率可达49.63%,B-1去除率可达62.34%。细菌N-1对NP的去除半衰期t1/2为41.44~48.02 h;B-1对BPA的去除半衰期t1/2为35.23~37.33 h。细菌N-1,B-1的最佳降解温度均为30℃,最佳p H值均在6.5~7.5之间,即两种细菌在中温、中性条件下对NP和BPA降解效果最佳。     

Metabolism of the Nonylphenol Isomer [Ring-U-14c]-4-(3′,5′-Dimethyl-3′-Heptyl)-Phenol by Cell Suspension Cultures of Agrostemma githago and Soybean

Abstract

The biotransformation of the nonylphenol isomer [ring-U-¹⁴C]-4-(3′,5′-dimethyl-3′-heptyl)-phenol (4-353-NP, consisting of two diastereomers) was studied in soybean and Agrostemma githago cell suspension cultures. With the A. githago cells, a batch two-liquid-phase system (medium/n-hexadecane 200:1, v/v) was used, in order to produce higher concentrations and amounts of 4-353-NP metabolites for their identification; 4-353-NP was applied via the n-hexadecane phase. Initial concentrations of [¹⁴C]-4-353-NP were 1 mg L^?1 (soybean), and 5 and 10 mg L^?1 (A. githago). After 2 (soybean) and 7 days (A. githago) of incubation, the applied 4-353-NP was transformed almost completely by both plant species to four types of products: glycosides of parent 4-353-NP, glycosides of primary 4-353-NP metabolites, nonextractable residues and unknown, possibly polymeric materials detected in the media. The latter two products emerged especially in soybean cultures. Portions of primary metabolites amounted to 19–22% (soybean) and 21–42% of applied ¹⁴C (A. githago). After liberation from their glycosides, the primary 4-353-NP metabolites formed by A. githago were isolated by HPLC and examined by GC-EIMS as trimethylsilyl derivatives. In the chromatograms, eight peaks were detected which due to their mass spectra, could be traced back to 4-353-NP. Seven of the compounds were side-chain monohydroxylated 4-353-NP metabolites, while the remaining was a (side-chain) carboxylic acid derivative. Unequivocal identification of the sites of hydroxylation/oxidation of all transformation products was not possible. The main primary metabolites produced by A. githago were supposed to be four diastereomers of 6′-hydroxy-4-353-NP (about 80% of all products identified). It was concluded that plants contribute to the environmental degradation of the xenoestrogen nonylphenol; the toxicological properties of side-chain hydroxylated nonylphenols remain to be examined.     

Risk assessment of xenoestrogens in a typical domestic sewage-holding lake in China

Release of domestic sewage leads to accumulation of xenoestrogens in holding waters, especially in closed or semi-enclosed waters such as lakes. In the study, the occurrence, distribution, estrogenic activity and risk of eight xenoestreogens were evaluated in Lake Donghu, China. Nonylphenol (NP), octylphenol (OP), and bisphenol A (BPA) were identified as the main xenoestrogens ranging from tens of ng L⁻¹ (in the surface water) or ng g⁻¹ dw (in the suspended particles and sediment) to tens of μg L⁻¹ or μg g⁻¹ dw. The sum of 17β-estradiol equivalents (∑EEQs) ranged from 0.32 to 45.02 ng L⁻¹ in the surface water, 0.53 to 71.86 ng g⁻¹ dw in the suspended particles, and 0.09 to 24.73 ng g⁻¹ dw in the sediment. Diethylstilbestrol (DES) was determined as the main contributor to ∑EEQs followed by NP. The risk assessment showed a higher risk in the surface water than in the suspended particles and sediment in such domestic sewage-holding lake.     

Estrogen equivalent concentration of 13 branched para-nonylphenols in three technical mixtures by isomer-specific determination using their synthetic standards in SIM mode with GC-MS and two new diasteromeric isomers

Thirteen isomers of branched para-nonylphenols (para-NP) in three technical mixtures were isomer-specifically determined using their synthesized standards by SIM of structurally specific ions, m/z 135, 149 or 163 with GC–MS. Of the 13 isomers, four isomers, 4-(2,4-dimethylheptan-4-yl)phenol, 4-(4-methyloctan-4-yl)phenol, 4-(3-ethyl-2-methylhexan-2-yl)phenol (3E22NP) and 4-(2,3-dimethylheptan-2-yl)phenol synthesized for their determinations were first used as standard substances. The 13 isomers in the technical mixtures individually occurred at mass percent portion of more than 2%. The total mass percent portions in the mixtures from Tokyo Chemical Industry (TCI), Aldrich, and Fluka covered with 89 ± 2%, 75 ± 4% and 77 ± 2%, respectively. The abundance of 4-(3,6-dimethylheptan-3-yl)phenol in the three mixtures was the largest with 11.1 ± 2% to 9.9 ± 0.3%, while that of 4-(2-methyloctan-2-yl)phenol was the smallest with 2.9 ± 0.3% to 3.0 ± 0.2%. Additionally, structures of four new isomers of more than 1% portion present in a technical mixture were elucidated as two pairs of diastereomeric isomers: two types of 4-(3,4-dimethylheptan-4-yl)phenol (344NP) and those of 4-(3,4-dimethylheptan-3-yl)phenol (343NP). By estrogenic assay of 13 isomers with yeast estrogen screen system, the activity of 3E22NP was the highest, while that of 4-(3-methyloctan-3-yl)phenol was the least. Their relative activities to that of 3E22NP were individually calculated. Estrogenic equivalent concentrations of the three technical mixtures were predictively evaluated. The ratio of the EEC to the conventional concentration, total mass percent portions of the 13 isomers in technical mixtures were 0.208 for TCI, 0.206 for Aldrich and 0.205 for Fluka. The predicted estrogenic activity of measured concentration of para-NP in technical mixtures was approximately 5-fold greater than the measured estrogen agonist activity.     

3种增塑剂单独和联合暴露对MCF-7细胞增殖的影响

为了探究结构差异较大、应用较为广泛的几类增塑剂雌激素活性的联合效应,选择邻苯二甲酸二丁酯(DBP)、双酚A(BPA)和壬基酚(NP)作为邻苯二甲酸酯、双酚A类和烷基酚类增塑剂的代表物进行试验.用DBP、BPA和NP单独及两两混合处理MCF-7细胞.采用MTT法检测培养24h、48 h、72 h和96h时的细胞增殖情况.采用流式细胞术检测药物培养48h后的细胞生长周期分布,并计算细胞增殖指数(PI).运用效应叠加模型(ES)判定联合效应类型.结果表明,在单独暴露试验中,DBP、BPA和NP组PI均大于1,且均能提高S期(DNA合成期)细胞比例.因此,DBP、BPA和NP均能显著促进MCF-7细胞增殖.混合暴露试验中,1)DBP和BPA在MTT试验中24h、48h、72 h和96h时的效应叠加指数(ESI)分别为1.013 9、1.023 8、0.9999、1.010 8,在流式细胞仪试验中ESI为1.014 1.因此,DBP和BPA的雌激素活性联合效应为加和作用.2)DBP和NP在MTT试验中24 h、48 h、72 h和96 h时的ESI分别为1.004 0、1.008 6、1.011 5、1.010 3,流式细胞仪试验中ESI为0.997 0.因此,DBP和NP的雌激素活性联合效应为加和作用.3)BAP和NP在MTT试验中24h、48 h、72 h和96h时的ESI分别为0.980 6、0.981 8、0.977 7、0.973 3,流式细胞仪试验的ESI为0.912 8.由此可知,BPA和NP的雌激素活性联合效应为拮抗作用.因此,可以采用MCF-7细胞增殖试验研究环境污染物雌激素活性联合效应.     

壬基酚的环境生物地球化学研究进展及对新污染物管理的建议

近年来,我国大气环境、水环境和土壤环境质量得到一定的改善,“碧水蓝天”已经成为常态.但随着持久性有机污染物、内分泌干扰物、抗生素和微塑料等新污染物在环境中不断被检出,新污染物正逐步受到广泛的关注,壬基酚作为一种典型的内分泌干扰物也备受研究人员的关注.系统地概括了我国水体中壬基酚的环境行为和暴露水平,并基于风险商法和联合概率曲线法对壬基酚可能造成的生态风险进行了评估.结果表明,壬基酚对水生生物的毒性效应主要包括急性毒性、生长发育毒性、雌激素效应和繁殖毒性;壬基酚在我国主要流域水体中普遍存在,其浓度平均值在60～1 000 ng·L^-1,浓度最高值可达4 628 ng·L^-1;基于风险商法和联合概率曲线法的风险评估结果表明,壬基酚对我国主要流域里的水生生物均存在一定的风险.最后,总结了目前比较常用的壬基酚处理处置和风险管控技术,比较了国际上内分泌干扰物的监管方法,针对我国在新污染物环境管理中存在的问题,提出了针对性政策建议,研究结果可以为我国新污染物管理和管控提供参考.