3株细菌对土壤中芘和苯并芘的降解及其动力学

研究了3株多环芳烃(PAHs)高效降解菌对土壤中芘和苯并芘(BaP)的降解动态,用Michaelis-Menton和Monod动力学模型对结果进行拟合.结果表明,3株细菌对芘和BaP的降解率有显著性差异.芽孢杆菌(Bacillus sp.SB02)42 d对芘和BaP的降解率均最高.当土壤中芘和BaP的初始浓度为50 mg/kg时,芽孢杆菌(Bacillus sp.SB02)、动胶杆菌(Zoogloea sp.SB09)、黄杆菌(Flavobacterium sp.SB10)42 d对芘的降解率分别为42.69%、32.88%、25.07%,对BaP的降解率分别为33.04%、25.39%、22.02%.3株细菌对芘和BaP的降解速率也存在显著性差异.芽孢杆菌(Bacillus sp.,SB02)最快,1周可降解20.88%芘和12.6%的BaP,动胶杆菌(Zoogloea sp.SB09)次之,黄杆菌(Flavobacterium sp.SB10)降解速率最慢.

Tween80作用下苯并(a)芘的微生物降解研究

以具有致癌、致畸和致突变作用的苯并(a)芘(BaP)为目标污染物,利用多环芳烃高效将解菌-芽孢杆菌,研究了非离子型表面活性剂吐温80(TW-80)对BaP的增溶及生物降解过程的影响。结果表明:(1)通过TW-80促溶,BaP在水中的溶解度提高近20倍;(2)在BaP降解的过程中,TW-80亦能作为碳源被芽孢杆菌利用,不产生二次污染;(3)当BaP浓度为10 mg/L,TW-80浓度为500 mg/L,共代谢底物-琥珀酸钠为50 mg/L时,BaP及TW-80的降解效果最好;并且初步揭示了TW-80改变BaP的生物可利用性而促进其降解的微生物机理。

苯并(a)芘及其代谢产物的连续降解研究

在以驯化过的芽孢杆菌(BA-07)降解BaP的过程中,鉴定出2个BaP的未开环代谢产物顺式-4,5-二氢-4,5-二醇-BaP(cis-BP4,5-dihydrodiol)和顺式-7,8-二氢-7,8-二醇-BaP(cis-BP7,8-dihydrodiol).由于该产物对微生物有一定毒性,所以难于进一步降解.为提高BaP降解的同时,降低cis-BP4,5-dihydrodiol和cis-BP7,8-dihydrodiol的累积,对2种降解方法(即单纯用BA07降解和运用高锰酸钾与BA-07耦合的方法降解)进行了比较,并且优化了连续降解的参数.结果表明,①对BaP及其代谢产物的连续降解,化学氧化与微生物耦合(高锰酸钾与BA-07)的降解效果明显好于单纯利用微生物(细菌BA-07)的降解;②在同一时间取样,cis-BP4,5-dihydrodiol的残留率均高于cis-BP7,8-dihydrodiol;③当BaP的浓度为40μg/mL,培养基的最佳pH为7.0,以琥珀酸钠为共代谢底物,可以显著提高BaP降解率,降低cis-BP 4,5-dihydrodiol和cis-BP7,8-dihydrodiol的累积.同时提出了化学氧化与微生物协同的方法可以有效促进环境中持久有机污染物的连续降解.

低浓度苯并[a]芘诱导赤子爱胜蚓HSP70和HSP90转录上调研究

为寻找土壤低浓度多环芳烃污染分子生物标记物,采用了抑制消减双杂交的方法构建了赤子爱胜蚓在苯并[a]芘(BaP)人工土壤污染胁迫下的差异表达cDNA文库,经测序和基因比对分析后,在上调文库中分别发现2个与热休克蛋白HSP70和1个与HSP90显著匹配的cDNA克隆.经定量PCR验证了0.1mg.kg-1和1.0mg.kg-1BaP对赤子爱胜蚓HSP70和HSP90的诱导作用,表明这两个新克隆到的赤子爱胜蚓热休克蛋白基因可作为土壤污染监测的备选分子生物标记物.

共基质对10株细菌降解苯并芘的作用研究

从石油污染的污泥中分离驯化出10株细菌(SB01～SB10),利用生物摇床实验对其降解苯并芘(BaP)的效能进行试验,研究了有(或无)共基质(葡萄糖Glu,或菲PHE)对细菌降解BaP的影响,并采用ANOVA和Tukey多重比较进行分析。结果表明(,1)当以BaP为惟一碳源和能源且BaP初始浓度为50mg·L-1时(MS1),SB01的降解率最高,5d可降解31.0%;以Glu为共代谢基质时(MS2),SB09的降解率最高,可达36.9%;以PHE为共代谢基质时(MS3),SB01对BaP的降解率为46.0%。(2)Glu对SB01、SB02、SB03、SB07、SB10降解BaP有抑制作用,对SB01抑制作用最明显,使SB01的降解率降低了13.1%,Glu对SB05,SB08降解率无明显促进或抑制作用。(3)PHE对细菌降解BaP均表现出促进作用,对SB01的促进作用最明显,使其降解率提高15.0%。(4)Glu对SB09的促进作用大于PHE的促进作用,而对SB06,PHE的促进作用大于Glu。

混合菌固定化及其对土壤中芘和苯并芘的降解

从石油污染土壤中筛选出1株细菌(Bacillus sp.)和1株真菌(Mucor sp.),以12种不同材料为载体对混合菌进行固定化,研究了固定化混合菌的降解特性.结果表明,采用吸附法能有效实现混合菌在改性后蛭石上的固定化.制得的固定化混合菌,传质性能好,对芘(Pyr)和苯并芘(BaP)的降解率42 d分别可达94.96%和74.96%,明显高于游离菌对Pyr和BaP的降解率60.49%和50.09%.采用扫描电子显微镜(SEM)观察了固定化混合菌微观结构,同时探讨了固定化混合菌的传质过程

均匀设计在固定化毛霉载体配方优化中的应用

将均匀设计方法应用于固定化毛霉(Mucorsp.)载体配方优化中。在单因素实验基础上,采用U(1557)均匀设计表对影响毛霉生长的主要营养因子进行优化,利用SPSS建立了固定化毛霉对土壤中芘、苯并(a)芘(BaP)和总多环芳烃(PAHs)降解率的回归方程。结果表明,全相关系数分别达到0.999,0.997和0.997。而后通过无约束规划求解获得了固定化载体最佳配方:玉米芯92.1%,豆饼4.8%,CaSO41.6%,白糖1.4%,MgSO40.2%,有效地提高了固定化毛霉对土壤中PAHs的降解率,为将固定化微生物技术应用于非流体介质中PAHs污染的原位修复提供了可行途径,而且该载体配方用料来源广泛,成本低廉,工艺简单,安全无毒。

六株真菌对土壤中芘和苯并芘的降解及其动力学

研究了6株真菌对土壤中芘和苯并芘(BaP)的降解动态,用Michaelis-Menton和Monod动力学模型对结果进行拟合.结果表明,6株真菌对芘和BaP的降解速率有显著性差异,降解率相差不大.产黄青霉(Penicillium chrysogenum,SF04),在42d内对BaP的降解能力最强,可达71.31%,对芘的降解能力相对最弱.镰刀菌(Fusariumsp.,SF11),黑曲霉(Aspergillusniger,SF05),木霉(Trichodermasp.,SF02)和毛霉(Mucorsp.,SF06)42d对芘的降解率分别为86.22%,86.18%,85.41%,85.04%,对BaP的降解率分别为71.11%,69.44%,69.05%,69.72%.木霉(Trichodermasp.,SF02)和毛霉(Mucorsp.,SF06)对芘和BaP的降解速率均很快.

污染土壤中苯并(a)芘的微生物降解途径研究进展

苯并(a)芘(BaP)是一种具有强致癌、致畸和致突变的多环芳烃(PAHs)。为了修复BaP污染的土壤,探索其降解途径是很重要的。为此,综述了国内外有关污染土壤中苯并(a)芘的微生物降解情况,对不同真菌、细菌降解苯并(a)芘的能力、代谢途径、共代谢底物以及环境影响因素进行了介绍和比较,提出了苯并(a)芘中间代谢产物的累积及其环境毒性方面的研究是修复苯并(a)芘污染土壤的重要方向。

北京官厅水库流域农田地表径流生物可利用磷流失规律

在模拟降雨条件下(30-69mm/39min),对官厅水库流域玉米地和休闲地地表径流泥沙和生物可利用磷(BAP)流失进行了初步研究.累积泥沙产量受雨强、坡度和作物覆盖影响,变幅为305.1-24933.4g/10m2;径流平均颗粒态生物可利用磷(BPP)、溶解态磷(SP)浓度都超出水体允许临界值0.02mg/L,表明流域农田地表径流对库区水体存在潜在污染危害;径流累积BAP流失达0.08-4.804g/10m2,估算的农田径流BAP流失达0.49kg/(hm2.a)以上.79.7%以上的BAP是颗粒态的.

Towards the identification of the common features of bacterial biofilm development

UPNa. Instituto de Agrobiotecnología. Laboratorio de Biofilms Microbianos

Later Life Consequences of Developmental Mitochondrial DNA Damage in C. elegans

Mitochondria are responsible for producing the vast majority of cellular ATP, and are therefore critical to organismal health [1]. They contain thir own genomes (mtDNA) which encode 13 proteins that are all subunits of the mitochondrial respiratory chain (MRC) and are essential for oxidative phosphorylation [2]. mtDNA is present in multiple copies per cell, usually between 103 and 104 , though this number is reduced during certain developmental stages [3, 4]. The health of the mitochondrial genome is also important to the health of the organism, as mutations in mtDNA lead to human diseases that collectively affect approximately 1 in 4000 people [5, 6]. mtDNA is more susceptible than nuclear DNA (nucDNA) to damage by many environmental pollutants, for reasons including the absence of Nucleotide Excision Repair (NER) in the mitochondria [7]. NER is a highly functionally conserved DNA repair pathway that removes bulky, helix distorting lesions such as those caused by ultraviolet C (UVC) radiation and also many environmental toxicants, including benzo[a]pyrene (BaP) [8]. While these lesions cannot be repaired, they are slowly removed through a process that involves mitochondrial dynamics and autophagy [9, 10]. However, when present during development in C. elegans, this damage reduces mtDNA copy number and ATP levels [11]. We hypothesize that this damage, when present during development, will result in mitochondrial dysfunction and increase the potential for adverse outcomes later in life.

To test this hypothesis, 1st larval stage (L1) C. elegans are exposed to 3 doses of 7.5J/m2 ultraviolet C radiation 24 hours apart, leading to the accumulation of mtDNA damage [9, 11]. After exposure, many mitochondrial endpoints are assessed at multiple time points later in life. mtDNA and nucDNA damage levels and genome copy numbers are measured via QPCR and real-time PCR , respectively, every 2 day for 10 days. Steady state ATP levels are measured via luciferase expressing reporter strains and traditional ATP extraction methods. Oxygen consumption is measured using a Seahorse XFe24 extra cellular flux analyzer. Gene expression changes are measured via real time PCR and targeted metabolomics via LC-MS are used to investigate changes in organic acid, amino acid and acyl-carnitine levels. Lastly, nematode developmental delay is assessed as growth, and measured via imaging and COPAS biosort.

I have found that despite being removed, UVC induced mtDNA damage during development leads to persistent deficits in energy production later in life. mtDNA copy number is permanently reduced, as are ATP levels, though oxygen consumption is increased, indicating inefficient or uncoupled respiration. Metabolomic data and mutant sensitivity indicate a role for NADPH and oxidative stress in these results, and exposed nematodes are more sensitive to the mitochondrial poison rotenone later in life. These results fit with the developmental origin of health and disease hypothesis, and show the potential for environmental exposures to have lasting effects on mitochondrial function.

Lastly, we are currently working to investigate the potential for irreparable mtDNA lesions to drive mutagenesis in mtDNA. Mutations in mtDNA lead to a wide range of diseases, yet we currently do not understand the environmental component of what causes them. In vitro evidence suggests that UVC induced thymine dimers can be mutagenic [12]. We are using duplex sequencing of C. elegans mtDNA to determine mutation rates in nematodes exposed to our serial UVC protocol. Furthermore, by including mutant strains deficient in mitochondrial fission and mitophagy, we hope to determine if deficiencies in these processes will further increase mtDNA mutation rates, as they are implicated in human diseases.

Regulation and Economic Globalization: Prospects and Limits of Private Governance

http://www.bepress.com/bap/vol12/iss3/art11/

Influencia de distintas citocininas en el cultivo in vitro de apices caulinares de Fragaria x ananassa y F. virginiana

p.65-70

Retinal Pigment Epithelial Cell DNA is Damaged by Exposure to Benzo[a]pyrene, a Constituent of Cigarette Smoke.

This study examined the effect of exogenous benzo[ a ]pyrene (BaP), an important constituent of cigarette smoke, on cultured bovine retinal pigment epithelial (RPE) cells. Evidence is presented for its metabolic conversion into benzo[ a ]pyrene diol epoxide (BPDE) and the consequent formation of potentially cytotoxic nucleobase adducts in DNA. Cultured RPE cells were treated with BaP at concentrations in the range of 0–100 µm. The presence of BaP was found to cause inhibition of cell growth and replication. BaP induced the expression of a phase I drug metabolizing enzyme which was identified as cytochrome P450 1A1 (CYP 1A1) by RT–PCR and by Western blotting. Coincident with the increased expression of CYP 1A1, covalent adducts between the mutagenic metabolite BPDE and DNA could be detected within RPE cells by immunocytochemical staining. Additional support for their formation was afforded by nuclease P1 enhanced 32P-postlabelling assays on cellular DNA. Single-cell gel electrophoresis (comet) assays showed that exposure of RPE cells to BaP rendered them markedly more susceptible to DNA damage induced by broad band UVB or blue light laser irradiation. In the case of UVB, this is consistent with the photosensitization of DNA cleavage by nucleobase adducts of BPDE. Collectively, these findings imply that BaP has a significant impact on RPE cell pathophysiology and suggest mechanisms whereby exposure to cigarette smoke might cause RPE dysfunction and cell death, thus possibly contributing to degenerative disorders of the retina.