微囊藻毒素-LR降解菌的筛选及降解特性研究

从上海市淀山湖表层水体中筛选分离出了1株降解微囊藻毒素-LR(MC-LR)的细菌并研究了其降解特性。根据细胞形态结构、生理生化特征及其16S rDNA基因序列分析,鉴定分离菌株DHU-28(GenBank序列登录号为HM047512)属嗜麦芽寡养单胞菌(Stenotrophomonas maltophilia)。微囊藻毒素降解实验结果表明,该菌株能在以MC-LR为唯一碳源、氮源的无机盐培养基中生长,6 d内可将初始质量浓度为15 mg/L的MC-LR降解为8.12 mg/L,降解效率达到45.9%。菌株DHU-28的最适生长温度是30℃,最适生长pH为7.0。酵母粉、蛋白胨、葡萄糖等营养物质可以明显促进菌株对MC-LR的降解效率,尤其是加入50 mg/L酵母粉后,6 d降解率达到63.2%。     

微波辅助光催化降解水中酚的研究

利用改装的家用微波炉和自制的无电极灯(EDL)试验了5种酚在水溶液中的微波辅助光催化降解效果.结果表明,反应30 min,微波辅助光催化作用(MW/EDL/TiO2)能去除80%以上的苯酚、间硝基苯酚、对氯苯酚和对甲酚,相应溶液的总有机碳(TOC)均减少70%以上,2-萘酚的去除率为59%,溶液TOC减少54%;微波(MW或MW/TiO2)作用对酚的去除率有一定贡献;对上述5种酚的微波辅助光催化反应动力学进行了初步研究,发现均符合准一级动力学方程.     

N-乙烯基吡咯烷酮降解菌的筛选、降解特性及其竹炭固定化

从活性污泥中分离筛选得到1株N-乙烯基吡咯烷酮(NVP)高效降解菌株ZF1,根据菌株ZF1的形态特征、生理生化特性和16S rRNA基因序列同源性分析,将其初步鉴定为产脲节杆菌(Arthrobacter ureafaciens)。菌株ZF1能以NVP为唯一碳、氮源进行生长,并在60 h内完全降解200 mg·L~(-1)的NVP。菌株ZF1降解NVP的最适环境条件为温度30℃,初始pH 7.0,Na Cl浓度7 g·L~(-1)。为了更好地实现其在实际废水中的应用,将菌株ZF1固定到竹炭上,扫描电镜观察表明ZF1能很好的附着在竹炭表面,且固定菌对NVP的降解效率明显高于游离菌。游离菌和固定菌对NVP的降解均符合一级动力学模型。重复利用5次后,固定菌对NVP的降解率仍能达到98%。     

白腐菌S.commune降解微囊藻毒素-LR的研究

为研究真菌对微囊藻毒素的降解作用,以白腐菌S.commune为降解菌,微囊藻毒素-LR(MC-LR)为降解目标进行生物降解,考察了白腐菌预培养方式及降解过程中的培养方式、充氧方式、温度、初始pH以及MC-LR初始浓度对降解效果的影响.结果表明,白腐菌可有效降解MC-LR,经液体预培养白腐菌对MC-LR的降解效果好于固体预培养,白腐菌静置培养过程中每天充入纯氧1min有助于MC-LR的降解,白腐菌降解MC-LR的最佳初始pH为4.5,适宜温度为30～35℃.白腐菌对MC-LR的降解能力随MC-LR初始浓度的增加而降低.在最佳条件下,当MC-LR初始质量浓度为1 mg/L时,其完全降解需要2d;当MC-LR初始质量浓度为15 mg/L时,其完全降解需要7d.高浓度MC-LR(30 mg/L以上)会对白腐菌生长产生抑制作用.MC-LR降解中间产物的具体结构尚不清楚,有待未来深入分析研究.     

纺锤芽孢杆菌(Bacillus fusiformis)降解萘的特性及动力学

在前期研究中发现,纺锤芽孢杆菌(Bacillus fusiformis,BFN)可以用于降解水溶液的萘,为了解其降解过程,发现BFN菌生长量随着溶液中的萘的含量增加而提高。其中,萘的含量分别是30、50、100和200 mg/L时,BFN的生物量OD600值分别为0.057、0.081、0.126和0.193;降解培养基溶液COD的去除率分别为59.4%、65.3%、69.2%和70.6%,说明BFN菌在生长的过程中利用萘作为碳源。同时,动力学拟合发现,对不同含量萘的降解过程都符合一级降解动力学方程,且BFN菌的生长过程满足逻辑斯蒂方程。扫描电镜图表明,BFN菌在萘的存在下生长得更好。紫外光谱显示波长为276 nm的萘的吸收峰在降解后下降很多。红外光谱数据则表明,降解液中有2组新的吸收峰出现:一组出现在2 878、2 930和2 968 cm-1处,说明在萘的降解过程中有新的羧酸类生成;另一组出现在3 438、3 667和3 731 cm-1处有新的酚类物质生成。     

好氧反硝化苯酚降解菌的分离鉴定及动力学

从驯化菌群中分离筛选出一株好氧反硝化苯酚降解细菌,经生理生化反应及16S rDNA测序,鉴定为Diaphorobacter属细菌。在好氧条件下,该菌株以苯酚为唯一碳源和能源,利用NO-3-N作为反硝化电子受体,其生长与反硝化特性研究表明:在接种量5%(体积分数),30℃,180 r/min振荡培养条件下,菌株降酚能力可达1 400 mg/L,同时,能有效去除初始浓度为165 mg/L的硝酸盐氮,60 h其去除率为91.5%,高含量苯酚对菌体生长有一定的抑制作用。应用Haldane方程对其生长过程进行动力学模拟,拟合曲线与实验测定值相关性良好,各参数分别为μmax(最大比增长率)0.324 h-1,Ks(半饱和常数)9.36 mg/L,Ki(抑制常数)146.72 mg/L,通过理论分析及实验验证得,该菌株苯酚降解动力学与其生长动力学表现出相似的趋势。     

染料结晶紫降解菌分离及其特性研究

从印染废水中筛选分离得到结晶紫(CV)降解菌株H,并对其脱色条件和机理进行了研究.结果表明,在外加生长基质条件下,菌株H不仅对CV降解脱色而且能降解中间产物.在pH 7.0、35℃、摇床转速180 r/min条件下,菌株H有最佳的脱色效果.对于20 mg/L CV溶液,生长细胞降解0.5 h,脱色率可达95%.休眠细胞对CV的降解规律也进行了对比探讨.菌株H对CV脱色过程遵循一级动力学方程,脱色速率常数k随CV浓度升高而降低,生长细胞的k值约为休眠细胞的6～7倍.     

以萘为共代谢的芘的好氧生物降解

为更好有效去除地下水中的常见污染物芘,以萘为降解基质,利用苍白杆菌降解芘,并对反应影响因素进行了研究,模拟了反应动力学。结果表明:浓度为100 mg·L~(-1)、pH=7、25℃、萘的初始反应24 h后萘的去除率达到99.84%,芘的去除率达到37.5%。另外,菌株在萘初始浓度不同的条件下对萘的降解符合一级动力学,对芘的降解符合二级动力学。结果表明苍白杆菌在去除地下水中的萘和芘方面具有很大前景。     

光反应器中UV/Fenton光降解湖水中微囊藻毒素的研究

在光催化反应器中,采用UV/Fenton光催化氧化技术,对湖水中微囊藻毒素的光催化降解过程中各影响因子分析表明:微囊藻毒素光催化氧化降解率受光照强度、氧化剂种类及浓度、溶液pH和不同光催化体系等多因素的影响.光促催化氧化体系对MC-LR藻毒素降解具有显著的作用,在紫外光与氧化剂的协同作用下,紫外光照强度越高,越易于促进MC-LR藻毒素快速降解,降解率可到达80%以上.     

菌株Arthrobacter sp.DNS10降解酶固定化条件的响应面优化

以前期分离得到阿特拉津降解菌株Arthrobacter sp.DNS10为供试菌株,选取海藻酸钠作为固定化材料,采用响应面法优化该菌株所包含的降解酶的固定化条件。借助Plackett-Burman设计筛选确定海藻酸钠浓度、固定化体系的p H值、加酶量和Ca Cl2溶液质量分数4个因素作为影响固定化酶比酶活的典型因素。借助Box-Behnken设计及响应面回归分析拟合,确定适宜上述降解酶的最佳包埋固定化条件及方法为:在每10 m L海藻酸钠浓度为1.93%,p H为8.5的固定化基质中加入983μL的降解酶液(蛋白浓度为88μg·L-1),然后利用注射器将上述混合溶液滴加到质量分数为2.7%的Ca Cl2溶液中即可制备出比酶活最高的固定化酶,实际测定固定化酶的最优比酶活为0.190 2 U·mg-1(预测值为0.187 4U·mg-1)。上述固定化酶平均粒径约为(0.44±0.01)cm,其在连续6次的使用过程中比酶活仍能保持在初始值的77.5%以上。上述固定化处理可有效的改善降解酶的环境贮存特性,固定化酶在常温下保存35 d后比酶活仍能保持在其初始状态的12.34%,而游离酶则无活性检出。     

以汽油为底物好氧共代谢三氯乙烯

三氯乙烯(trichloroethylene,TCE)是土壤和地下水中广泛存在的有机污染物,好氧生物降解因可将污染物彻底转化成无毒的终产物,一直受到广泛关注,但是TCE好氧降解需要共代谢底物。首次提出以汽油为底物,选取真养产碱杆菌作为活性降解菌株,对地下水中三氯乙烯的好氧共代谢降解进行了初步研究。分别优化了共代谢底物、底物与TCE浓度比、培养基、pH值、盐度、溶解氧等条件,确定了最佳降解条件。当水中TCE的浓度为1 mg/L时,通过对体系预曝氧气,调节汽油浓度为10 mg/L,pH值为5,降解24 h,TCE的降解率可达66.8%。为修复同时被汽油和TCE污染的场地提供了一个新的研究方向。     

Biodegradation of s-triazine herbicide atrazine by Enterobacter cloacae and Burkholderia cepacia sp. from long-term treated sugarcane-cultivated soils in Kenya

In this study soils from sugarcane-cultivated fields were screened for bacterial species capable of atrazine (6-chloro-N²-ethyl-N⁴-isopropyl-1,3,5-triazine-2,4-diamine) degradation due to long exposure of the soils to this herbicide. To enrich for atrazine degraders, Minimal Salt Medium containing atrazine as the sole N source and glucose as the C source was inoculated with soils impacted with this herbicide and incubated. Bacterial growth was monitored by measuring optical density. The degradation of atrazine was followed by measuring residual atrazine in liquid cultures over a given time period by high performance liquid chromatography. Bacterial strains isolated from the enrichment cultures were characterized by biochemical tests and identified by 16S rRNA gene sequencing. Two bacterial strains coded ISL 8 and ISL 15 isolated from two different fields were shown to have 94 and 96% 16S rRNA gene sequence similarity to Burkholderia cepacia respectively. Another bacterial sp., ISL 14 was closely related to Enterobacter cloacae with a 96% 16S rRNA gene sequence similarity. There was not much difference between the extents of atrazine degradation by the enrichment cultures with communities (79–82% applied amount) from which pure strains were isolated and the pure strains themselves in liquid cultures that showed a degradation of 53–83% of applied amount. The study showed existence of bacterial strains in different sugarcane-cultivated fields which can use atrazine as a nitrogen source. The bacterial strains isolated can be used to enhance the degradation of atrazine in contaminated soils where atrazine is still considered to be recalcitrant.     

Influence of pH on the sonolysis of ciprofloxacin: Biodegradability, ecotoxicity and antibiotic activity of its degradation products

The presence of antibiotics in the aquatic environment has raised concerns due to the potential risk for the emergence or persistence of antibiotic resistance. Antibiotics are often poorly degraded in conventional wastewater treatment plants. In this study, sonolysis at 520 kHz and 92 W L⁻¹ was used for the degradation of the fluoroquinolone antibiotic ciprofloxacin. In a first experiment at pH 7, 57% of the ciprofloxacin (15 mg L⁻¹) was degraded after 120 min of ultrasonic irradiation at 25 °C. pH proved to be an important parameter determining the degradation rate, since the pseudo first order degradation constant increased almost fourfold when comparing treatment at pH 7 (0.0058 min⁻¹) and pH 10 (0.0069 min⁻¹) with that at pH 3 (0.021 min⁻¹). This effect can be attributed to the degree of protonation of the ciprofloxacin molecule. The BOD/COD ratio of the solutions, which is a measure for their biodegradability, increased from 0.06 to 0.60, 0.17, and 0.18 after 120 min of irradiation depending on the pH (3, 7, and 10, respectively). The solution treated at pH 3 can even be considered readily biodegradable (BOD/COD > 0.4). The antibiotic activity against Escherichia coli (G−) and Bacillus coagulans (G+) of the treated solutions also reduced after sonolysis. The highest decrease was again found when irradiated at pH 3. In contrast, ecotoxicity of the solutions to the alga Pseudokirchneriella subcapitata increased 3- to 10-fold after 20 min of treatment, suggesting the formation of toxic degradation products. The toxicity slowly diminished during further treatment.     

Neurotoxicological evaluation of microcystin-LR exposure at environmental relevant concentrations on nematode Caenorhabditis elegans

Previous studies have not examined the adverse effects of microcystin-LR (MC-LR) at environmental relevant concentrations on the development and functions of nervous system. The neurotoxic effects of MC-LR exposure on neurotransmitter systems were investigated in Caenorhabditis elegans. After exposing L1 larvae to 0.1, 1, 10, and 100 μg?l^?1 of MC-LR for 8 and 24 h, the adverse effects on GABAergic, cholinergic, serotonergic, dopaminergic, and glutamatergic neurons were examined. The expression levels of genes required for development and functions of GABAergic neurons were further investigated. Body bend frequency and head thrash frequency decreased significantly after MC-LR exposure for 8 h at concentrations more than 1 μg?l^?1 and after MC-LR exposure for 24 h at concentrations more than 0.1 μg?l^?1. Loss of GABAergic neurons increased significantly in a dose-dependent manner after MC-LR exposure at concentrations more than 0.1 μg?l^?1. In contrast, no obvious neuronal losses or morphologic changes were observed in cholinergic, serotonergic, dopaminergic, and glutamatergic neurons in MC-LR-exposed nematodes. Quantitative real-time PCR assay further showed that expression levels of unc-30, unc-46, unc-47, and exp-1 genes required for development and function of GABAergic neurons decreased significantly in nematodes exposed to MC-LR at concentrations more than 0.1 or 1 μg?l^?1. MC-LR at environmental relevant concentrations caused neurobehavioral defects, which may be largely due to the neuronal loss and the alterations of expression level of genes required for GABAergic neurotransmitter system in C. elegans.     

Detection of free microcystins in the liver and muscle of freshwater fish by liquid chromatography-tandem mass spectrometry

MC analysis of biological tissue is considered to be very difficult due to the lack of validated methods. This is the primary limiting factor for monitoring potential risks in both the flesh of aquatic organisms and the aquatic ecosystem. In this study, an effective method to determine free MCs (MC-LR and MC-RR) in the muscle and liver tissues of freshwater cultured fish was developed using solid-phase extraction (SPE) and liquid chromatography-tandem mass spectrometry (LC/MS-MS). The extraction solvent, time of extraction, eluent and purification of the extract were optimized. Various SPE cartridges were also investigated. In this optimized analytical procedure, an 85% methanol/water solution (v/v) was selected as the extraction solvent, after which the extracts were purified by removing fats and proteins; a HLB cartridge was chosen for MCs enrichment; and 90% methanol containing 0.02% formic acid/water solution (v/v) was used as the eluent. Under the optimized pretreatment conditions and instrument parameters, good recoveries of MC-LR and MC-RR were obtained at three concentrations (0.5, 1.0 and 2.0 µg g^?1 dry weight (DW)), with values ranging from 92.5 to 98.3% and 92.1 to 98.6%, respectively. The method detection limit (MDL) for muscle samples was 0.5 µg kg^?1 and 0.4 µg kg^?1 (DW) for MC-LR and MC-RR, respectively. The MDL for the liver samples was 0.8 µg kg^?1 (DW) for both MC-LR and MC-RR. The developed procedure was successfully applied to analyze MCs in the muscle and liver of fish samples collected from a Chinese freshwater aquaculture pond during bloom seasons. The MC-LR concentrations ranged from below the MDL to 4.17 µg kg^?1 and the MC-RR concentrations ranged from below the MDL to 2.64 µg kg^?1.     

脱氮副球菌YF1微生物燃料电池生物阴极脱氮和产电

以脱氮副球菌YF1构建纯种生物阴极微生物燃料电池(microbial fuel cell,MFC)进行脱氮和产电机理的研究。研究结果发现,阴极碳氮比、pH值对产电和脱氮效率有明显影响。当MFC的阴极运行条件pH值为8.0,碳氮比为20时,运行时间15 h时,脱氮率高达100%,输出电压为150 mV。上述结果表明,微生物燃料电池运行过程中,细菌降解硝酸根的机理为将硝酸根还原为N2或者直接将其作为自身的营养物质而利用。循环伏安(CV)与扫描电镜(SEM)的结果表明,在微生物燃料电池运行中,副球菌YF1通过接触导电作为产电的电子供体。     

Endophytic bacteria improve phytoremediation of Ni and TCE co-contamination

The aim of this work was to investigate if engineered endophytes can improve phytoremediation of co-contaminations by organic pollutants and toxic metals. As a model system, yellow lupine was inoculated with the endophyte Burkholderia cepacia VM1468 possessing (a) the pTOM-Bu61 plasmid, coding for constitutive trichloroethylene (TCE) degradation, and (b) the ncc-nre Ni resistance/sequestration system. Plants were exposed to Ni and TCE and (a) Ni and TCE phytotoxicity, (b) TCE degradation and evapotranspiration, and (c) Ni concentrations in the roots and shoots were determined. Inoculation with B. cepacia VM1468 resulted in decreased Ni and TCE phytotoxicity, as measured by 30% increased root biomass and up to 50% decreased activities of enzymes involved in anti-oxidative defence in the roots. In addition, TCE evapotranspiration showed a decreasing trend and a 5 times higher Ni uptake was observed after inoculation.     

Effects of cyanobacteria producing microcystins on seed germination and seedling growth of several agricultural plants

The effects of cyanobacteria aqueous extracts containing Microcystin-LR (MC-LR) on the seed germination and growth of Pisum sativum, Lens esculenta, Zea mays and Triticum durum were investigated. Experiments were carried out on a range of doses of the extract (equivalent to 0, 1.6, 2.9, 5.8, 8.7 and 11.6 μ g MC-LR/mL). The results confirm that these plants were sensitive to cell-free extracts of a toxic Microcystis and that germination inhibition was dose dependent. One-way analysis of variance (ANOVA) showed that P. sativum is the most sensitive tested species with a 97% germination rate reduction and L. esculenta was the most resistant. At the 8th day, the exposure to the microcystins (MC) resulted in a significant decrease of plant epicotyls length, roots length and a net inhibition of lateral root formation. It is concluded that MC could affect also terrestrial plants seedling germination and growth. Therefore, the use of water for irrigation contaminated by MC could exert negative biochemical effects on seed and plant metabolism which might influence the agricultural crops.     

木糖氧化无色杆菌及混合菌群对多环芳烃的降解特性

采用木糖氧化无色杆菌及混合菌降解水中多环芳烃。考察了木糖氧化无色杆菌的降解广谱性及其对多环芳烃混合底物的降解,特别考察了混合菌对具有弱致癌性的■(Chrysene)的降解特性。结果表明,木糖氧化无色杆菌具有较宽的降解谱,对多环芳烃混合底物具有良好降解特性。当蒽、菲、芘和■4种PAHs共存时,木糖氧化无色杆菌对蒽、菲、芘和■的降解效率分别达83%、66%、85%和80%。与单一木糖氧化无色杆菌相比,混合菌对的降解效率较高。尖镰孢菌与木糖氧化无色杆菌、茄镰孢菌与木糖氧化无色杆菌和3株菌同时共存时,■的降解效率分别达87%、88%和86%。