Evaluation of Bacillus sp. MZS10 for decolorizing Azure B dye and its decolorization mechanism

To evaluate decolorization and detoxification of Azure B dye by a newly isolated Bacillus sp. MZS10 strain, the cultivation medium and decolorization mechanism of the isolate were investigated. The decolorization was discovered to be dependent on cell density of the isolate and reached 93.55%(0.04 g/L) after 14 hr of cultivation in a 5 L stirred-tank fermenter at 2.0 g/L yeast extract and 6.0 g/L soluble starch and a small amount of mineral salts. The decolorization metabolites were identified with ultra performance liquid chromatography-tandem mass spectroscopy(UPLC-MS). A mechanism for decolorization of Azure B was proposed as follows: the C=N in Azure B was initially reduced to –NH by nicotinamide adenine dinucleotide phosphate(NADPH)-dependent quinone dehydrogenase, and then the –NH further combined with –OH derived from glucose to form a stable and colorless compound through a dehydration reaction. The phytotoxicity was evaluated for both Azure B and its related derivatives produced by Bacillus sp. MZS10 decolorization, indicating that the decolorization metabolites were less toxic than original dye. The decolorization efficiency and mechanism shown by Bacillus sp. MZS10 provided insight on its potential application for the bioremediation of the dye Azure B.     

Effects of glucose on the decolorization of Reactive Black 5 by yeast isolates

The cometabolic roles of glucose were investigated in decolorization of an azo dye, Reactive Black 5, by yeast isolates, Debaryomyces polymorphus and Candida tropicalis. The results indicated that the dye degradation by the two yeasts was highly associated with the yeast growth process and glucose presence in the medium. Color removal of 200 mg dye/L was increased from 76.4% to 92.7% within 60 h to 100% within 18-24 h with the increase of glucose from 5 to 10 g/L, although the activity of manganese dependent peroxidase （MnP） decreased by 2-8 times in this case. Hydrogen peroxide of 233.3 μg/L was detected in 6 h in D. polymorphus culture. The cometabolic functions of glucose and hydrogen peroxide could be also confirmed by the further color removals of 95.8% or 78,9% in the second cycle of decolorization tests in which 7 g glucose/L or 250 μg H202/L was superadded respectively together with 200 mg dye/L.     

Emissions of fluorides from welding processes

The levels of fluoride airborne particulates emitted from welding processes were investigated. They were sampled with the patented IOM Sampler, developed by J. H Vincent and D. Mark at the Institute of Occupational Medicine(IOM), personal inhalable sampler for simultaneous collection of the inhalable and respirable size fractions. Ion chromatography with conductometric detection was used for quantitative analysis. The efficiency of fluoride extraction from the cellulose filter of the IOM sampler was examined using the standard sample of urban air particle matter SRM-1648 a. The best results for extraction were obtained when water and the anionic surfactant N-Cetyl-N-N-Ntrimethylammonium bromide(CTAB) were used in an ultrasonic bath. The limits of detection and quantification for the whole procedure were 8 μg/L and 24 μg/L, respectively The linear range of calibration was 0.01–10 mg/L, which corresponds to 0.0001–0.1 mg of fluorides per m3 in collection of a 20 L air sample. The concentration of fluorides in the respirable fraction of collected air samples was in the range of 0.20–1.82 mg/m3, while the inhalable fraction contained 0.23–1.96 mg/m3 of fluorides during an eight-hour working day in the welding room.     

Degradation of 2,2′,4,4′-tetrabromodiphenyl ether by Pycnoporus sanguineus in the presence of copper ions

The degradation of 2,2′,4,4′-tetrabromodiphenyl ether(BDE-47) by Pycnoporus sanguineus was investigated in order to explore the impact of the heavy metal Cu~(2+) on BDE-47 decomposition and the subsequent formation of metabolites, as well as to further elucidate the degradation mechanism of BDE-47. An increase in degradation rate from 18.63% to49.76% in the first four days and its stabilization at(51.26 ± 0.08)% in the following days of BDE-47 incubation were observed. The presence of Cu~(2+) at 1 and 2 mg/L was found to promote the degradation rate to 56.41% and 60.79%, respectively, whereas higher level of Cu~(2+) (≥ 5 mg/L) inhibited the removal of BDE-47. The similar concentration effects of Cu~(2+) was also found on contents of fungal protein and amounts of metabolites. Both intracellular and extracellular enzymes played certain roles in BDE-47 transportation with the best degradation rate at 27.90% and 27.67% on the fourth and third day, individually. During the degradation of BDE-47, four types of hydroxylated polybrominated diphenyl ethers(OH-PBDEs), i.e., 6′-OH-BDE-47, 5′-OH-BDE-47, 4′-OH-BDE-17, 2′-OH-BDE-28, and two bromophenols, i.e., 2,4-DBP and 4-BP were detected and considered as degradation products. These metabolites were further removed by P. sanguineus at rates of 22.42%,23.01%, 27.04%, 27.96%, 64.21%, and 40.62%, respectively.     

Polybrominated diphenyl ether (PBDE) in blood from children (age 9–12) in Taizhou, China

Polybrominated diphenyl ethers (PBDEs) as ubiquitous persistent organic pollutants have attracted much attention in recent years. Exposure to PBDEs could induce a high health risk for children. The aim of this study was to investigate the PBDEs exposure of children (9–12 years) from Taizhou, China. Fifty-eight blood samples were collected in one school in a mountainous area in Taizhou. The concentrations of Σ9PBDEs (sum of BDE-28, -47, -99, -100, -153, -154, -183, -197 and -209) ranged from 2.66 to 33.9 ng/g lipid wet (lw) with a median of 7.22 ng/g lw. These concentrations were lower than those of children in USA, but close to European and Asian general population levels. The results showed that children in Taizhou countryside were at a low level of PBDEs exposure. The predominant congener was BDE-209, followed by BDE-28, -47, -197 and -153. High abundance of BDE-209 was consistent with the pollution background of PBDEs in China characterized by high brominated congeners as main pollutants.     

Continuous treatment of azo acid dyes by photo-dependent denitrifying sludge

Simultaneous removals of dye and nitrate by photo-dependent denitrifying sludge(PDDS)have been demonstrated in a continuousflow bench-scale reactor.The best C/N for the degradation of azo dyes by PDDS was 1.5.The specific removal rate of azo dye AB92 decreased with a decrease in hydraulic retention time and increased with a decrease in solids retention time.The degradation rate of TOC decreased with a decrease in hydraulic retention time.AB92,which has nitro and hydroxyl substitutions in non-para positions,was uniquely degraded.During continuous flow treatment experiments using PDDS,complete degradation of azo dyes AB92 and AO20 at influent concentrations of 40mg/L and 30mg/L,respectively,was achieved with an HRT of 16.     

Dispersive liquid–liquid microextraction of silver nanoparticles in water using ionic liquid 1-octyl-3-methylimidazolium hexafluorophosphate

Using the ionic liquid(IL)1-octyl-3-methylimidazolium hexafluorophosphate as the extractant and methanol as the dispersion solvent,a dispersive liquid–liquid microextraction method was developed to extract silver nanoparticles(AgN Ps)from environmental water samples.Parameters that influenced the extraction efficiency such as IL concentration,pH and extraction time were optimized.Under the optimized conditions,the highest extraction efficiency for AgN Ps was above 90% with an enrichment factor of 90.The extracted AgN Ps in the IL phase were identified by transmission electron microscopy and ultraviolet–visible spectroscopy,and quantified by inductively coupled plasma mass spectrometry after microwave digestion,with a detection limit of 0.01 μg/L.The spiked recovery of AgN Ps was 84.4% with a relative standard deviation(RSD)of 3.8%(n = 6)at a spiked level of 5 μg/L,and 89.7% with a RSD of 2.2%(n = 6)at a spiked level of 300 μg/L,respectively.Commonly existed environmental ions had a very limited influence on the extraction efficiency.The developed method was successfully applied to the analysis of Ag NPs in river water,lake water,and the influent and effluent of a wastewater treatment plant,with recoveries in the range of 71.0%–90.9% at spiking levels of 0.11–4.7 μg/L.     

A monoclonal antibody against Lates calcarifer vitellogenin and a competitive ELISA to evaluate vitellogenin induction after exposure to xenoestrogen

A monoclonal antibody specific to sea bass(Lates calcarifer) vitellogenin(VTG) was developed,for use as a tool for monitoring endocrine disrupting chemicals(EDCs). VTG was induced in sea bass by intramuscular injection of 17β-estradiol(E_2: 2 mg/kg) every three days. Blood was collected three days after the last injection. Plasma VTG was then purified by chromatography in hydroxyapatite and a sephacryl-S300 column. Characterizations of purified VTG were done by phospholipoglycoprotein staining on a native-PAGE with confirmation by mass spectrometry(LC-MS/MS). Antibody was raised in mice by injection of purified VTG. After monoclonal antibody production, the hybridoma clone No. 41(MAb-sea bass VTG 41)was selected and developed for quantification of VTG by competitive enzyme-linked immunosorbent assay(ELISA). The ELISA method was sensitive with a detection limit of VTG 40 ng/mL. MAb-sea bass VTG 41 was specific to VTG from E_2-treated sea bass and others EDCs(Nonylphenol, Benzo[a]pyrene and CdCl_2). Moreover, cross-reactivity was also found in E_2-treated coral grouper(Epinephelus corallicola). The ELISA method obtained from this work can be further applied for the assessment of EDCs in Thailand and Southeast Asia's aquatic environment.     

Characteristics of gas-liquid pulsed discharge plasma reactor and dye decoloration efficiency

The pulsed high-voltage discharge is a new advanced oxidation technology for water treatment. Methyl Orange (MO) dye wastewater was chosen as the target object. Some investigations were conducted on MO decoloration including the discharge characteristics of the multi-needle reactor, parameter optimization, and the degradation mechanism. The following results were obtained. The color group of the azo dye MO was effectively decomposed by water surface plasma. The decoloration rate was promoted with the increase of treatment time, peak voltage, and pulse frequency. When the initial conductivity was 1700 μS/cm, the decoloration rate was the highest. The optimum distance between the needle electrodes and the water surface was 1 mm, the distance between the grounding electrode and the water surface was 28 mm, and the number of needle electrodes and spacing between needles were 24 and 7.5 mm, respectively. The decoloration rate of MO was affected by the gas in the reactor and varied in the order oxygen > air> argon > nitrogen, and the energy yield obtained in this investigation was 0.45 g/kWh.     

Enhanced anaerobic fermentation with azo dye as electron acceptor： Simultaneous acceleration of organics decomposition and azo decolorization

Accumulation of hydrogen during anaerobic processes usually results in low decomposition of volatile organic acids（VFAs）. On the other hand, hydrogen is a good electron donor for dye reduction, which would help the acetogenic conversion in keeping low hydrogen concentration. The main objective of the study was to accelerate VFA composition through using azo dye as electron acceptor. The results indicated that the azo dye serving as an electron acceptor could avoid H2 accumulation and accelerate anaerobic digestion of VFAs. After adding the azo dye, propionate decreased from 2400.0 to 689.5 mg/L and acetate production increased from 180.0 to 519.5 mg/L. It meant that the conversion of propionate into acetate was enhanced. Fluorescence in situ hybridization analysis showed that the abundance of propionate-utilizing acetogens with the presence of azo dye was greater than that in a reference without azo dye. The experiments via using glucose as the substrate further demonstrated that the VFA decomposition and the chemical oxygen demand（COD） removal increased by 319.7 mg/L and 23.3% respectively after adding the azo dye. Therefore, adding moderate azo dye might be a way to recover anaerobic system from deterioration due to the accumulation of H2 or VFAs.     

BDE-47对2种海洋微藻光合特性的影响

研究了不同浓度的2,2′,4,4′-四溴联苯醚(BDE-47)对2种海洋微藻的光合特性的影响. 结果表明, 在试验所设定的浓度范围内 (0.1~2.5μg/L),海水小球藻和赤潮异弯藻的PSII最大光能转化效率(Fv/Fm)、光化学淬灭系数(qP)未受到显著抑制,表明2种微藻PSII反应中心在试验过程中未受到损伤.当BDE-47浓度为2.5μg/L时,海水小球藻的PSII潜在活性(Fv/Fo)、PSII实际光能转化效率(φPSII)和光合电子传递效率(ETR)在96h受到显著抑制,而赤潮异湾藻的3个参数在暴露期间均未受到抑制,表明赤潮异湾藻对BDE-47的耐受性强于海水小球藻.各叶绿素荧光参数中, Fv/Fo、φPSII、ETR更适合作为指示海洋环境中BDE-47污染水平的生物标志物.     

5种多溴联苯醚同系物对海洋饵料藻(亚心型扁藻和盐生杜氏藻)的急性毒性

本研究选择5种多溴联苯醚同系物(BDE-28、BDE-47、BDE-99、BDE-153和BDE-209)对2种海洋饵料藻(亚心型扁藻和盐生杜氏藻)进行急性毒性试验。96 h半抑制浓度(96 h EC50)计算结果表明,BDE-28 和BDE-47的毒性较高,随溴原子取代数的增加,PBDE同系物对海洋饵料藻的毒性呈下降趋势,5种PBDE同系物对亚心形扁藻、盐生杜氏藻的96 h EC50由小及大依次为:BDE-28(128,75 g/L)、BDE-47(114,120 g/L)、BDE-99(383,572 g/L)、BDE-153(996,1249 g/L)和 BDE-209(2056,1868 g/L)。目前全球近岸海水中PBDEs浓度尚不会对海洋饵料藻产生急性毒性。海洋微藻对同种PBDE同系物的敏感性差异较大(96 h EC50相差1~2个数量级),为客观评价PBDEs的海洋生态风险性,应研究PBDEs对更多种类微藻的毒性效应。     

Effects of nitrogen and phosphorus concentrations on the bioaccumulation of polybrominated diphenyl ethers by Prorocentrum donghaiense

The growth, cellular total lipids, bioaccumulation amount, and bioaccumulation factors (BAFs) of 2,4,4 ′ -tribromodiphenyl ether (BDE28), 2,2 ′ ,4,4 ′ -tetrabromodiphenyl ether (BDE47), and 2,2 ′ ,4,4 ′ ,5-pentabromodiphenyl ether (BDE99) in a semi-continuous culture of Prorocentrum donghaiense were studied in relation to nitrate (0, 128, and 512 μmol/L) and phosphate (0, 8, and 32 μmol/L) concentrations. The BDE28, BDE47, and BDE99 content per cell under 0 μmol N/L were 3.77 × 10 6 , 3.95 × 10 6 , and 4.32 × 10 6 ng/cell, respectively, which were significantly higher than those under 128 and 512 μmol N/L. A nearly 5-fold increase in polybrominated diphenyl ether (PBDE) content per algal cell was found between 0 and 8 μmol P/L and between 8 and 32 μmol P/L. With increasing N and P concentrations, the PBDE content per volume of algal culture and the accumulation percentage of available PBDEs declined slightly. The BAFs for the PBDEs based on lipids showed that the logBAF lip under 0 μmol N/L was higher than those under 128 and 512 μmol N/L. The logBAF lip under 0 μmol P/L was higher than that under 8 μmol P/L but lower than that under 32 μmol P/L. Correlation analysis indicated a significant negative correlation between nutrient concentration and cellular total lipids, as well as the PBDE content per cell. The results indicate that different N and P concentrations change the total lipids content of P. donghaiense, thereby resulting in varying PBDE accumulation.     

BDE-47在紫贻贝中的分布、蓄积、消除和毒性效应

以紫贻贝(Mytilus galloprovincialis)为试验生物,研究了 2,2',4,4'-四溴联苯醚(BDE-47)在贻贝组织中的分布、生物蓄积和消除动态,并探究BDE-47对贻贝的毒性作用.结果发现:紫贻贝对BDE-47有较强的生物蓄积能力和一定的消除能力,且蓄积具有组织特异性和浓度依赖性,消化腺和鳃是B...     

剑尾鱼卵黄蛋白原的ELISA检测

以卵黄脂磷蛋白(lipovitellin,Lv)抗血清为抗体,以纯化的卵黄蛋白原(vitellogenin,Vtg)为抗原,建立了间接酶联免疫吸附反应(ELISA)方法检测剑尾鱼(Xiphophorus helleri)雄鱼整体匀浆液中ρ(Vtg).结果表明,该方法的检测灵敏度为7.8 ng/mL,批内变异系数为5.094%,批间变异系数为5.540%,工作范围为32.5～2 000 ng/mL,在该范围内,标准曲线具有良好的线性和重复性.由于该方法可直接在1块或在不同的酶标板上准确地进行比较,因而可利用ELISA方法测定经诱导雄鱼整体匀浆液中ρ(Vtg)水平.结果显示,50 μg/L 17-β雌二醇(E₂)诱导21 d的雄性剑尾鱼有明显的Vtg产生;10 μg/L E₂诱导剑尾鱼亦有Vtg产生,但较50 μg/L E₂诱导组低;1 μg/L E₂诱导组及对照组剑尾鱼没有Vtg产生,检测孔OD450/对照OD₄₅₀(P/N)值小于2.1.      

厌氧条件下 2,2',4,4'-四溴联苯醚的微生物降解

多溴联苯醚是环境中的新兴污染物,其中2,2’,4,4’-四溴联苯醚(BDE-47)在环境(尤其是水环境)中普遍存在且生物毒性很高.以BDE-47为对象,研究了6组含脱卤球菌的培养液对BDE-47的降解,目的是了解厌氧条件下BDE-47的微生物降解及其动力学.采用100 mL血清瓶作为厌氧反应器,对厌氧微生物进行培养.对每组菌作两种处理,一是仅加入BDE-47作为能源(设计终浓度为200μg.L-1),另一是同时加入BDE-47和三氯乙烯(TCE)作为能源(设计终浓度分别为200μg.L-1和13mg.L-1).经过3个月的实验,两组含脱卤球菌的培养液(6M6B和T2)均能明显降解BDE-47,生成BDE-17、BDE-4及少量的DE,TCE的存在一定程度上减弱了6M6B和T2菌对BDE-47的降解.采用PCR-DGGE法对不同培养菌液的群落结构进行比较,发现醋酸杆菌属与BDE-47的降解关联较大.在3种不同初始浓度(50、250和500μg.L-1)条件下,BDE-47的降解速率分别为0.003 3、0.001 4和0.001 0 d-1.本研究表明,厌氧条件下BDE-47在细菌的作用下可发生还原降解,生成BDE-17和BDE-4.醋酸杆菌属可能在BDE-47的降解中起较大作用.高浓度的BDE-47在一定程度上会抑制降解菌的活性.     

猪肝微粒体体外代谢2,2’,4,4’-四溴联苯醚

多溴联苯醚(PBDEs)是一类在环境中普遍存在的持久性有机污染物,研究PBDEs的体外代谢行为对理解其在体内的富集和转化具有重要意义。文章以猪肝微粒体作为研究对象,以体外代谢形式研究其对2,2’,4,4’-四溴联苯醚(BDE-47)的代谢能力,优化了代谢条件,并研究了0.01、0.05、0.1、0.5、1、5、10 mg/L的BDE-47对细胞色素P450酶系中7-乙氧基香豆素-O-脱乙基酶(ECOD)、7-乙氧基异吩唑酮-O-脱乙基酶(EROD)和苯胺4-羟基化酶(ANH)活性的影响。结果表明,孵育0.1 mg/L的BDE-47时,猪肝微粒体对BDE-47的代谢率可达到28.6%。实验所设各浓度BDE-47均能够保护微粒体的ECOD活性,但较高浓度的BDE-47(0.5、1、5 mg/L)对微粒体的EROD活性有显著抑制作用,而各浓度BDE-47对ANH活性没有显著影响。     

2,4-D完全抗原的合成及其免疫性能评价

以2,4-D为半抗原,BSA为载体蛋白,通过EDC的偶联反应条件优化试验,合成了多种结合比的完全抗原,制备了小分子环境污染物的多克隆抗体.结果表明,偶联反应优化步骤是2,4-D、BSA和EDC同时反应,4°C条件下反应18h.偶联反应可以在2,4-D的浓度为10.0～12.0 mg/mL的0.05mol/L的磷酸盐缓冲溶液中进行,反应缓冲体系pH控制在5.4～6.1之间.EDC的加入量低于12 mg时,采用2,4-D和多聚赖氨酸的偶联物为包被抗原.通过balb/c小鼠免疫试验评价结合比分别为6、12、18和25的完全抗原的免疫性能,结果表明结合比为12和18的完全抗原具有很好的免疫原性.其中以结合比为18的完全抗原免疫小鼠获得的抗血清对包被抗原载体的非特异性吸附低于阴性血清,2,4-D特异性血清含量高,适于作为完全抗原免疫小鼠进一步制备单克隆抗体.     

Role of sorbent surface functionalities and microporosity in 2,2’,4,4’-tetrabromodiphenyl ether sorption onto biochars

The study provides insight into the combined effect of sorbent surface functionalities and microporosity on2,2 ′,4,4 ′-tetrabromodiphenyl ether (BDE-47) sorption onto biochars. A series of biochars prepared underdifferent conditionswere used to test their sorption behaviorswith BDE-47. The extents of sorption behaviorswere parameterized in terms of the single-point adsorption equilibrium constant (Koc ) at three equilibrium concentration (C e ) levels (0.001Sw (solubility), 0.005Sw , and 0.05Sw )whichwasdetermined using the Freundlich model. To elucidate the concentration-dependentdominant mechanisms for BDE-47 sorption onto biochars, K ocwas correlatedwith four major parameters using multiple parameter linear analysis accompaniedwith significance testing. The results indicated that at low concentration (Ce = 0.001Sw ), the surface microporosity term,which represented a pore-filling mechanism, contributed significantly to this relationship,while as concentrationwas increased to higher levels, surface functionality related to surface adsorption began to take thedominant role,whichwas further confirmed by the results of Polanyi-based modeling. Given the above results, adual mode model based on Dubinin-Radushkevich andde Boer-Zwikker equationswas adopted to quantitatively assess the changes of significance of surface adsorption aswell as that of pore fillingwith sorption processdevelopment. In addition, UV spectra of four typical aromatic compoundswhich represented the key structural fragments of biochars before and after interactionswith BDE-47were analyzed todetermine the active functional groups and supply complementary evidence for thedominant interaction force for surface adsorption, based onwhich π-π electron-donor-acceptor interactionwas proposed to contribute greatly to surface adsorption.     

重金属对秀丽隐杆线虫繁殖的影响

为了探究重金属单独及联合暴露对秀丽隐杆线虫(Caenorhabditis elegans)繁殖的影响及其作用机制,采用96孔板液相暴露试验,以环境相关浓度(0.1~50.0 μg/L)的Cd、Hg、Pb暴露同步化处理秀丽隐杆线虫,24 h后统计其怀卵数及阴门畸形率,以及连续记录72 h内子一代秀丽隐杆线虫的个体数,最后利用2×2析因设计方差分析阐明重金属的两两联合作用模式. 结果表明:与对照组相比,Cd、Hg、Pb单独及联合暴露对秀丽隐杆线虫的繁殖具有显著抑制作用,ρ(Cd)为50.0 μg/L、ρ(Hg)为10.0 μg/L、ρ(Pb)为50.0 μg/L暴露组秀丽隐杆线虫子一代数量分别降低了43.5%、33.8%、51.0%;Cd-Hg暴露组〔当ρ(Cd)、ρ(Hg)分别为50.0、10.0 μg/L时〕、Hg-Pb暴露组〔当ρ(Hg)、ρ(Pb)分别为10.0、50.0 μg/L时〕、Cd-Pb暴露组〔当ρ(Cd)、ρ(Pb)分别为50.0、50.0 μg/L时〕秀丽隐杆线虫子一代个体数分别降低了44.7%、61.0%、53.3%. 进一步研究发现,Cd、Hg、Pb单独及联合暴露会引起秀丽隐杆线虫子宫内受精卵数量降低以及产卵器阴门结构畸形率升高. 2×2析因设计方差分析结果发现,Cd、Hg、Pb两两联合暴露对秀丽隐杆线虫的繁殖表现为没有交互作用或者交互方式为拮抗. 研究显示,秀丽隐杆线虫子宫内怀卵数的降低是其子一代数量显著减少的主要原因,而阴门结构的损伤可能使产卵行为受损,从而进一步加剧重金属对秀丽隐杆线虫繁殖的抑制作用.