高效原油降解菌筛选驯化实验研究

由含油废水原有微生物种群中分离出纯种菌株，通过纯种菌株的单一菌种降解性能实验，混合菌株垢正交性能实验，筛选出7种高效原油降解菌（包括芽孢杆菌属，黄杆菌属，黄单胞菌属，和假单胞菌属），其中芽孢杆菌属降解原油能力最强，对于18．4mg/L的含油废水，降解去除率高达94％－95％，实验还表明，单种菌株降解原油的能力比混合菌株要好。     

原油降解菌的筛选鉴定及降解特性的研究

实验从河北省唐山市柳赞镇冀东油田油井污水中筛选出8株以原油为唯一碳源的降解菌株(编号为T1-8),通过形态学特征观察以及16SrDNA分子生物学鉴定,确定这8个菌株分别为铜绿假单胞菌(Pseudomonas aeruginosa)、金黄色葡萄球菌(Staphylococcusaureus)、Bacillus safensis、短小芽孢杆菌(Bacillus pumilus)、嗜冷杆菌(Psychrobacter pulmonis)、鲁氏不动杆菌(Acinetobacter lwoffii)、纺锤芽孢杆菌(Bacillus fusiformis)和洋葱伯克霍尔德氏菌(Burkholderia cepacia)。采用无机盐培养基,以华北任丘油田原油为唯一碳源,分别对上述8种菌株进行原油降解能力测试,通过为期7d的降解,其相对应的降解率分别为67.14%、43.13%、30.37%、24.74%、43.00%、37.94%、55.28%和14.51%,而且其中的金黄色葡萄球菌和嗜冷杆菌对原油具有降解能力为首次报导。后续对铜绿假单胞菌进行了影响降解率的单因素实验,实验结果显示在pH为7、初始原油浓度为5g/L和菌液接种量1mL(约1.8×1010cfu/mL)时铜绿假单胞菌对原油的降解率达到其最大值67.14%。     

PAHs降解菌的分离、鉴定及降解能力测定

以芴、菲、蒽、芘为碳源和能源筛选、分离PAHs降解菌。14株能降解利用PAHs的菌株被分离。通过HPLC分析,在含芴、菲、蒽、芘的混合培养基质中10号菌的降解能力最强。研究它的降解性能和生长情况,表明该菌在混合反应体系中培养30d后对芴、菲、蒽、芘的降解率分别为95.27、90.46、28和80%;在只含一种PAH的单反应体系中该菌对芴、菲、蒽的降解能力提高,降解率分别可达98.91、94.32和52.17%,而对芘的降解能力则降低,降解率仅为62.47%。与混合PAHs培养体系相比,在单一PAH培养体系中,细菌的对数生长期缩短1/3。经生理生化鉴定和16SrDNA序列对比分析,确定10号菌株属于假单胞菌,命名为PseudomonasspFAP10。     

紫外诱变对假单胞菌生物特性的影响

室温下,用紫外光对假单胞菌株进行2 min诱变。分别用驯化后紫外诱变和未诱变假单胞菌处理系列浓度(20、100、200、300、400 mg/L)苯酚水样,对比分析其在降解速率、降解时间、微生物呼吸率、酶活性等降解特性方面的差异性。结果表明:假单胞菌经紫外诱变后,对苯酚的耐受性得以提高,去除高浓度苯酚的速率得以提高,处理较高浓度苯酚的能力明显提升。诱变假单胞菌完全降解较高浓度苯酚的时间比未诱变假单胞菌短,诱变假单胞菌对于高浓度苯酚的适应性比未诱变假单胞菌更好。苯酚降解过程中,脱氢酶始终保持着较高的活性,假单胞菌经诱变后其酶活性得以提高。     

硝基苯好氧降解的共基质及生物协同作用

通过研究多菌种多基质条件下硝基苯的降解以便寻找提高实际废水处理效率的途径,从不同菌源中筛选与驯化获得硝基苯的高效降解菌枯草芽孢杆菌、门多萨假单孢菌、肺炎克雷伯氏菌及人苍白杆菌.研究这些菌种好氧降解硝基苯过程中的生物协同作用及其基质效应,构建实际废水处理的生物降解体系．结果表明,枯草芽孢杆菌和人苍白杆菌之间存在协同降解作用．加入人苍白杆菌后可使质量浓度为232．0mg/L硝基苯被完全降解的时间从145h缩短为85h;由枯草芽孢杆菌、门多萨假单孢菌和肺炎克雷伯氏菌组成的混合降解实验发现,枯草芽孢杆菌和门多萨假单孢菌之间存在一定程度的相互抑制作用;当加入肺炎克雷伯氏菌后抑制作用消失,降解速度加快.复合菌的作用明显优于各菌单独作用或任何两种菌之间的配合效果.经氯霉素制药厂含硝基苯实际废水的检验,混合菌在降解硝基苯的同时也使COD成分得到有效去除．     

不同菌源的微生物对邻苯二甲酸二辛酯生物降解性的比较

从处理石化厂废水的活性污泥中分离出 1株荧光假单胞菌FS 1及处理焦化厂废水的活性污泥中分离出 1株铜绿假单胞菌FS 2和 1株短杆菌FS 3。研究了这 3株菌对邻苯二甲酸二辛酯的最适降解条件 ,比较了其降解特性。荧光假单胞菌FS 1、铜绿假单胞菌FS 2和短杆菌FS 3最适pH分别为 6 5～ 8 0 ,7 0～ 8 0和 7 0～ 8 0 ;温度为 2 0～ 3 5℃ ,1 5～ 3 5℃和 1 5～ 3 5℃。他们对邻苯二甲酸二辛酯的降解反应符合一级动力学特征 ,其半寿期 :荧光假单胞菌FS 1 <铜绿假单胞菌FS 2 <短杆菌FS 3 ;荧光假单胞菌FS 1是一株高效的邻苯二甲酸二辛酯降解菌。 更多还原      

细菌协同降解阴离子型聚丙烯酰胺的机理

为了探究细菌混合协同降解阴离子型聚丙烯酰胺（HPAM）机理,作者以球红假单胞菌和枯草芽孢杆菌等体积混合构成混合菌,研究了单个菌株和混合菌的生长情况及降解特性。同时采用分子对接模拟了红球菌N-771酰胺酶（Rh Amidase）和枯草芽孢杆菌漆酶(Lac)与HPAM结构模型的结合。试验结果表明,枯草芽孢杆菌含有内生孢子,其适应环境的能力比球红假单胞菌强,而且2种菌株都含有鞭毛,运动剧烈。在温度35℃、pH 7、接种量2 mL和7 d的最佳条件下混合菌的降解率为39.24%,而球红假单胞菌和枯草芽孢杆菌的降解率分别为24.9%和22.13%。分子对接结果表明,Rh Amidase-HPAM-2亲和力最大、最稳定的主要原因是强氢键作用,而且它们的结合最佳。与Lac相比,Rh Amidase更容易攻击短链HPAM的酰胺侧链而引发水解。而Lac可以容纳一定长度的HPAM的碳链,倾向于氧化HPAM的碳链。根据试验结果与分子对接模拟的相互佐证,提出了细菌混合降解HPAM协同的机理,实质上是Rh Amidase和Lac共同催化降解HPAM。     

焦化废水中氨氮降解菌降解性能的研究

本实验从焦化废水中分离出一株高效氨氮降解菌A1,经过生理生化鉴定,初步确定为假单胞杆菌菌属。本实验对该菌种的降解性能进行研究,确定该菌种的最佳降解条件:p H8.0,温度30℃,菌种量5%(V菌悬液:V培养基),在此条件下,对200mg/L的氨氮去除率达到89.5%。     

生物净水工艺中降解有机污染物优势菌的筛选初探

为了获得高效有机污染物降解的优势菌株,以稳定运行的生物净水工艺为研究对象,采用细菌分离、生化鉴定、脱氢酶活性分析、净水效能测定和降解相关基因扩增等方法对生物滤池中微生物群落进行了分析。结果表明,从生物滤池中分离的优势菌主要为巨大芽胞杆菌、短小芽胞杆菌、蜡样芽胞杆菌、弗氏枸橼酸杆菌、豚鼠气单胞菌、恶臭假单胞菌和肺炎克雷氏菌等。在8株高脱氢酶活性的菌株中,1株巨大芽孢杆菌、1株蜡样芽孢杆菌和2株恶臭假单胞菌对原水中的高锰酸盐指数UV254和TOC的去除效率较高;并从1株恶臭假单胞菌株的基因组DNA中成功扩增出大小为548 bp的偏三苯酚1,2-双加氧酶基因片段(对硝基苯酚降解相关基因),说明该菌株具有降解对硝基苯酚污染物的潜能。因此,本研究可筛选高效有机污染物降解的优势菌株,为生物净水工艺提供优良的菌种资源。     

油烟污染物降解优势菌的驯化培育及其性能

通过驯化污水厂曝气池中的活性污泥,获得以细菌、真菌为主的降解油烟污染物的优势菌种.驯化21d后,细菌的数量由16000cfu/mL减少到1500cfu/mL,种类由8种减少为2种.真菌包含丝状真菌和酵母状真菌,且数量显著增加.对驯化后的菌种进行形态学、生理学和分子生物学鉴定,结果为芽孢杆菌(Bacillussubtilis)、铜绿假单胞菌(Pseudomonasaeraginosa)、青霉菌(Penicillum)和红冬孢酵母菌(Rhodosporidiumtoruloides).在30℃,初始油烟污染物浓度为70.6mg/L条件下,铜绿假单胞菌、芽孢杆菌、青霉菌、红冬孢酵母菌的最大油烟比降解率分别为0.627,0.916,2.102,2.453mg/g菌,优势菌群降解能力最高可达到7mg/g菌.     

青顶拟多孔菌对单一和复合多环芳烃的降解特性

利用中国东北林区普遍存在白腐菌——青顶拟多孔菌,降解单一和复合多环芳烃,分别测定了菲、蒽、芘于11,22,33d的累积降解率.结果显示,对于单一多环芳烃,该菌种降解能力由强到弱依次为菲>蒽>芘,33d累积降解率依次为96.56%、94.76%和57.53%;对复合多环芳烃降解中,菲和芘的累积降解率分别为99.46%和61.09%.在复合多环芳烃的降解研究中发现,少量蒽的加入,刺激了菌种对菲和芘的降解,使菲和芘的降解率分别提高了2.9%和3.56%.由此提示,在研究降解高环、难降解多环芳烃时,可利用低环多环芳烃对菌种的刺激作用,在体系内形成高、低环多环芳烃的共代谢,以达到更加高效降解多环芳烃的目的.     

固相微萃取和固相萃取评价多环芳烃降解过程中的生物有效性变化

为了揭示多环芳烃(PAHs)降解过程中的生物有效性变化规律,应用PAHs降解菌剂对PAHs污染焦化厂土壤进行微生物修复,采用固相微萃取及固相萃取方法提取评价PAHs的生物有效性,分析降解率和生物有效性变化的差异及相关关系.结果表明,该焦化厂土壤中PAHs总量以低环PAH为主,微生物菌剂对土壤总PAHs降解率为68.3%;微生物作用后,孔隙水中PAHs的降低以3、4环为主,孔隙水中PAHs变化率普遍低于土壤中的降解率;Tenax-TA提取降解前后土壤中的PAHs,降解后3、4环PAHs的快速解吸组分降低,5、6环的快速解吸组分变化不大;土壤中PAHs降解量分别与PAHs孔隙水浓度和Tenax-TA快速提取量存在相关关系.以上结果表明可用PAHs孔隙水浓度以及Tenax-TA快速提取量预测微生物对土壤PAHs的降解,这为焦化厂土壤PAHs污染的修复提供了理论依据.     

降解石油丝状真菌的筛选及其降解特性

从大庆石油污染土壤中分离得到14株石油降解丝状真菌,用以筛选可高效降解石油的菌群. 经过ITS(转录间隔区序列)分析,其中9株属于镰孢霉属(Fusarium sp.),2株属于黄白生丛赤壳菌属(Bionectria sp.),另外3株分别属于葡萄穗酶菌属(Stachybotrys sp.)、曲霉属(Aspergillus sp.)和雅致放射毛霉属(Actinomucor sp.);在固体培养基中各菌株的生长速率差异显著(P<0.01). 不同菌株能够特异降解不同的石油组分. 将筛选的优势菌株组成2个菌群,菌群1由菌株3、D2、D3和D52组成,菌群2由菌株3、6、D2和D3组成,分别考察单菌和2个菌群对原油的降解效果,结果表明:菌株原油降解试验30d后,单菌菌株D52对原油去除率最大,为64.25%;菌群1和菌群2对原油的去除率较高,分别达到74.55%和72.64%,可以考虑用于污油生物修复治理的工程菌群开发研究.      

Adaptability of microbial inoculators and their contribution to degradation of mineral oil and PAHs

Five dominant bacteria strains（A cetobacter sp., Alcdigenes sp., Micrococcus sp., Arthrobacter sp. and Bacillus sp.） and five fungi strains （Cephalosporium sp. Ⅰ, Cephalosporlum sp. Ⅱ, Aspergillus sp. Ⅰ, Aspergillus sp. Ⅱ and Fusarium sp.） isolated from petroleum-contaminated soil were used to assess the potential capability of mineral oil and PAH enhanced degradation separately and jointly using the batch liquid medium cultivation with diesel oil spiked at 1000 mg/L. The experiment was performed on a reciprocal shaker in the darkness at 25℃ to 30℃ for 100 d. The dynamic variation in the activity of microbial inoculators in each treatment and the degradation of the target pollutants during the period of experiment were monitored. Results showed a more rapid biodegradation of mineral oil and PAHs at the beginning of the experiment （about 20 d） by dominant bacteria, fungi and their mixture than that of the indigenous microorganisms, however, thereafter an opposite trcnd was exhibited that the removal ratio by indigenous microorganisms was superior to any other dominant treatments and the tendency lasted till the end of the experiment, indicating the limited competitive capability of dominant microorganisms to degrade the contaminants, and the natural selection of indigenous microorganisms for use in the removal of the contaminants. At the end of the experiment, the removal ratio of mineral oil ranged from 56.8 % to 79.2 % and PAHs ranged from 96.8 % to 99.1% in each treatment by microbial inoculators,     

Degradation of pyrene by immobilized microorganisms in saline-alkaline soil

Biodegradation of polycyclic aromatic hydrocarbons (PAHs) is very difficult in saline-alkaline soil due to the inhibition of microbial growth under saline-alkaline stress. The microorganisms that can most effectively degrade PAHs were screened by introducing microorganisms immobilized on farm byproducts and assessing the validity of the immobilizing technique for PAHs degradation in pyrene-contaminated saline-alkaline soil. Among the microorganisms examined, it was found that Mycobacterium sp. B2 is the best, and can degrade 82.2% and 83.2% of pyrene for free and immobilized cells after 30 days of incubation. The immobilization technique could increase the degradation of pyrene significantly, especially for fungi. The degradation of pyrene by the immobilized microorganisms Mucor sp. F2, fungal consortium MF and co-cultures of MB+MF was increased by 161.7% (P < 0.05), 60.1% (P < 0.05) and 59.6% (P < 0.05) after 30 days, respectively, when compared with free F2, MF and MB+MF. Scanning electron micrographs of the immobilized microstructure proved the positive effects of the immobilized microbial technique on pyrene remediation in saline-alkaline soil, as the interspace of the carrier material structure was relatively large, providing enough space for cell growth. Co-cultures of different bacterial and fungal species showed different abilities to degrade PAHs. The present study suggests that Mycobacterium sp. B2 can be employed for in situ bioremediation of PAHs in saline-alkaline soil, and immobilization of fungi on farm byproducts and nutrients as carriers will enhance fungus PAH-degradation ability in saline-alkaline soil.     

Efficient degradation of lube off by a mixed bacterial consortium

A laboratory study was performed to assess the biodegradation of lube oil in bio-reactor with 304# stainless steel as a biofilm carrier.Among 164 oil degrading bacterial cultures isolated from oil contaminated soil samples, Commaonas acidovorans Pxl, Bacillus sp.Px2, Pseudomonas sp. Px3 were selected to prepare a mixed consortium for the study based on the efficiency of lube oil utilization.The percentage of oil degraded by the mixed bacterial consortium decreased slightly from 99% to 97.2% as the concentration of lube oil was increased from 2000 to 10,000 mg/L. The degradation of TDOC (total dissolved organic carbon) showed a similar tendency compared with lube oil removal, which indicated that the intermediates in degradation process hardly accumulated. Selected mixed bacterial consortium showed their edge compared to activated sludge. Scanning electron microscopy (SEM) photos showed that biofilms on stainless steel were robust and with a dimensional framework constructed by EPS (extracellular polymeric substances),which could promote the biodegradation of hydrocarbons. The increase of biofilm followed first-order kinetics with rate of 0.216 μg glucose/(cm~2·day) in logarithm phase. With analysis of Fourier transform infrared spectroscopy (FT-IR) and gas chromatography-mass spectrometry (GC-MS) combined with removal of lube oil and TDOC, mixed bacterial consortium could degrade benzene and its derivatives, aromatic ring organic matters with a percentage over 97%.     

砂土中柴油的微生物降解研究

从加油站污染土壤中筛选出对柴油具有较强降解能力的菌株,比较研究菌株及菌群降解柴油的差异,探讨植物苜蓿和芥菜对菌株降解柴油污染物的影响.结果表明:①菌株Q18和菌群对砂土中柴油都具有较强的降解能力,菌群对柴油的降解率明显高于菌株Q18.菌株Q18 和菌群在5 d内对柴油的总降解率分别为40.29%和54.15%.与菌株Q18降解柴油相比,菌群主要是强化了中、长链烷烃化合物的降解.②在砂土中,苜蓿和芥菜都能显著地强化菌株Q18对柴油的降解,但苜蓿强化菌株Q18降解柴油的能力强于芥菜. 5 d内菌株Q18-芥菜和菌株Q18-苜蓿复合体系对柴油总降解率分别达到60.05%和56.68%.③菌群和苜蓿及芥菜复合体系降解柴油的能力也有显著提高,但芥菜强化菌群降解柴油的能力强于苜蓿.5 d内菌群-芥菜和菌群-苜蓿复合体系对柴油总降解率分别为75.53%和70.50%.      

Efficient degradation of lube oil by a mixed bacterial consortium

A laboratory study was performed to assess the biodegradation of lube oil in bio-reactor with 304# stainless steel as a biofilm carrier. Among 164 oil degrading bacterial cultures isolated from oil contaminated soil samples, Commaonas acidovorans Px1, Bacillus sp. Px2, Pseudomonas sp. Px3 were selected to prepare a mixed consortium for the study based on the efficiency of lube oil utilization. The percentage of oil degraded by the mixed bacterial consortium decreased slightly from 99% to 97.2% as the concentration of lube oil was increased from 2000 to 10,000 mg/L. The degradation of TDOC (total dissolved organic carbon) showed a similar tendency compared with lube oil removal, which indicated that the intermediates in degradation process hardly accumulated. Selected mixed bacterial consortium showed their edge compared to activated sludge. Scanning electron microscopy (SEM) photos showed that biofilms on stainless steel were robust and with a dimensional framework constructed by EPS (extracellular polymeric substances), which could promote the biodegradation of hydrocarbons. The increase of biofilm followed first-order kinetics with rate of 0.216 μg glucose/(cm2·day) in logarithm phase. With analysis of Fourier transform infrared spectroscopy (FT-IR) and gas chromatography-mass spectrometry (GC-MS) combined with removal of lube oil and TDOC, mixed bacterial consortium could degrade benzene and its derivatives, aromatic ring organic matters with a percentage over 97%.     

松花江干流PAHs的底泥-水交换行为及时空异质性

为了解松花江干流底泥和水体中PAHs(多环芳烃)的环境分布行为,应用逸度方法研究了松花江中PAHs的底泥-水交换行为及时空异质性特征. 结果表明:K_OW(辛醇-水分配系数)影响PAHs的底泥-水交换行为,并与底泥-水交换的f_f(逸度分数)呈显著负相关(R=-0.801,P=0.000),而f_f与PAHs的溶解度则呈正相关(R=0.499,P<0.05);高环PAHs的底泥-水交换行为受w(OC)变化的影响较为强烈,w(OC)每提高0.10%,2~6环PAHs的f_f降低0.7%~11.0%;春季PAHs的底泥-水交换的f_f大于夏季. 低环的Nap(萘)表现出明显的由底泥向水体的迁移行为,Phe(菲)和FlA(荧蒽)几乎接近于平衡状态,而高环的BaP(苯并芘)和BgP(苯并苝)则相反. 夏季PAHs的大气传输及本地排放源的沉降,可能为松花江干流PAHs的主要来源;汇入支流的输入可视为主干河流水体中污染物的另一来源. 水体中2~4环PAHs处于中等变异,5~6环PAHs则表现为强变异;底泥中3~4环PAHs处于中等变异,而其他环数PAHs则呈强变异. 从季节性变化来看,夏季底泥中PAHs的CV(变异系数)相对较大,而春季水体中PAHs的CV则略大于夏季. 研究显示,PAHs物理化学性质的差异,水体中悬浮颗粒物和底泥中w(OC),以及外源性PAHs的输入,均会使不同环数PAHs在水体和底泥中的CV产生较大差异.      

焦化废水A/O2和A/O/H/O处理工艺中多环芳烃的削减行为分析

多环芳烃(PAHs)是焦化废水的特征污染物,不同的焦化废水处理工艺会影响其归趋行为与削减量.基于此,本文追踪了焦化废水厌氧/好氧/好氧(A/O~2)和厌氧/好氧/水解/好氧(A/O/H/O)处理工艺中PAHs的分布行为,使用气质联用色谱(GC-MS)检测并分析了两种工艺中各单元反应器内废水及污泥样品的PAHs浓度,通过通量衡算,评估工艺系统处理效果的差异性.结果表明,两种工艺的前置A厌氧池对PAHs的削减不明显,PAHs在A池富集的浓度远高于O池及H池,低环PAHs经水相从A池进到O池后得到有效降解,高环PAHs则大部分富集于污泥相中,风险比较大,应该区别处理;A/O/H/O工艺对低环的PAHs生物削减率略高于A/O~2工艺,对4环以上的PAHs,A/O/H/O则表现出更明显的降解有效性,理解为A/O/H/O工艺的H池促进了大分子有机物的水解,对PAHs具有针对性.综合研究结果认为,A/O/H/O工艺表现出比A/O~2更明显的降解PAHs的优势.