浑河底泥产甲烷古菌多样性空间分布特征

为了深入探究影响河流甲烷排放的关键因素,采用PCR-RFLP(限制性片段长度多态性聚合酶链反应)技术及测序分析对浑河底泥产甲烷古菌的多样性及空间分布进行了研究.结果表明,浑河底泥产甲烷古菌划分为4大类群:甲烷杆菌目(Methanobacteriales)、甲烷微菌目(Methanomirobiales)、甲烷胞菌目(Methanocellales)和甲烷八叠球菌目(Methanosarcinales).从河流上游到下游,产甲烷古菌的群落结构呈现出空间变异的特征.河流上游占优势的产甲烷古菌菌属为Methanosarcina和Methanospirillum,分别约占46.2%和20.8%;中游主要菌属为Methanosarcina、Methanobacterium和Methanomethylovorans,分别约占40.9%、20.7%和15.2%;下游主要菌属为Methanosarcina和Methanobacterium,分别约占46.7%和22.1%.浑河底泥中Methanosarcina为优势菌属.Shannon指数(H')和Simpson多样性指数(D)计算结果表明,河流上游(H'=1.56,D=0.33)和中游(H'=1.79,D=0.26)产甲烷古菌的多样性与下游(H'=1.65,D=0.32)均较为接近.     

Microbial community changes in aquifer sediment microcosm for anaerobic anthracene biodegradation under methanogenic condition

The widespread distribution of polycyclic aromatic hydrocarbons (PAHs) in groundwater has become an important environmental issue. Knowledge of microbial community changes could aid in identification of particular microorganisms that are capable of degrading PAHs in contaminated aquifers. Therefore, 16S rRNA gene clone library analysis was used to identify the archaeal and bacterial communities in an aquifer sediment microcosm used for anaerobic anthracene degradation under methanogenic conditions. A remarkable shift of the archaeal community structure occurred after anaerobic anthracene degradation, but the types of the abundant bacterial phyla did not change. However, a decrease of both archaeal and bacterial diversity was observed. Bacterial genera Bacillus, Rhodococcus and Herbaspirillum might have links with anaerobic anthracene degradation, suggesting a role of microbial consortia. This work might add some new information for understanding the mechanism of PAH degradation under methanogenic conditions.     

互花米草沼泽湿地产甲烷古菌的多样性及垂向分布

为了深入探究影响河口沼泽湿地甲烷排放空间变异的关键因素,采用PCR-RFLP技术及测序分析对闽江河口互花米草沼泽湿地产甲烷古菌的多样性及垂向分布进行了研究.系统发育分析表明,闽江河口互花米草沼泽湿地产甲烷古菌的多样性划分为3大类群:甲烷杆菌目(Methanobacteriales)、甲烷微菌目(Methanomirobiales)和甲烷八叠球菌目(Methanosarcinales).在不同的土壤深度,产甲烷古菌的群落结构呈现出空间变异的特征.0~10 cm土层占优势的产甲烷古菌菌属为Methanoregula和Methanobacterium,分别约占46%和43%;10~20 cm土层主要菌属为Methanoregula、Methanobacterium和Methanolobus,分别约占66%、13%和12%,Methanoregula为优势菌属;20~30 cm土层主要菌属为Methanoregula、Methanobacterium和Methanogenium,分别约占65%、11%和12%,Methanoregula为优势菌属.Shannon指数(H')和Simpson多样性指数(D)计算结果表明,10~20 cm土层(H'=2.69,D=0.073)和20~30 cm土层(H'=2.47,D=0.093)产甲烷古菌的多样性明显高于0~10 cm土层(H'=1.60,D=0.292).     

微生物修复油污土壤过程中氮素的变化及菌群生态效应

对陕北地区石油污染土壤进行了微生物修复研究,利用气相色谱-质谱联用技术(gas chromatography-mass spectrometry,GC-MS)测定土壤中烷烃和多环芳烃的含量变化,采用高通量测序技术研究微生物修复对油污土壤微生物群落多样性的影响,对修复过程中土壤不同形态氮含量变化进行了分析测定.结果表明,经过18周的修复,土壤中烷烃含量由初始时的25 987.8mg·kg~(-1)降低为12 788.6 mg·kg~(-1),多环芳烃含量由初始时的5 322.9 mg·kg~(-1)降低为2 917.2mg·kg~(-1).高通量测序结果表明,经过微生物修复处理的土壤微生物群落结构发生了较大变化,一些可降解烃类的菌群如厚壁菌门(Firmicutes)、拟杆菌门(Bacterodetes)、迪茨氏菌属(Dietzia)、不动杆菌属(Acinetobacter)所占丰度增加.经过修复处理的土壤中总氮、氨氮含量呈降低趋势,硝态氮含量在修复前期呈降低趋势,修复后期基本保持不变.研究结果表明,微生物修复处理可去除土壤中的大部分烷烃和多环芳烃,烷烃和多环芳烃的降解效果与土壤中硝氮含量和烃降解菌所占丰度相关.     

天津表土PAHs区域环境风险评价研究

在参照国外相关标准基础上,应用指示克立格方法对天津地区表层土壤PAHs进行区域环境风险评价,并对评价结果进行比分析.结果表明,按照美国风险评价标准,只有组分Bap的含量超标,风险区域仅为8.12%和2.34%;按照荷兰风险评价标准,除了Ant和Baa的风险区域所占天津地区总面积的比重比较小之外(分别为5.26%和68.42%),其余所占比重都非常大,全部超过90%以上;按照加拿大风险评价标准,除了Nap的风险区域所占比重比较大(为97.89%)、Phe和Pyr所占比重中等(分别为56.84%和47.65%)之外,其余参与风险评价组分的风险区域所占比重都比较小,全部都在5%以下;按照丹麦(西兰岛)风险评价标准,只有组分Nap超标,且风险区域仅为9.26%.通过不同国家评价标准结果的比较,能充分了解天津地区目前土壤PAHs风险特征,对于制定符合天津地区特征以及符合中国特征的土壤PAHs风险评价标准具有重要的意义.     

Impacts of produced water origin on bacterial community structures of activated sludge

The purpose of this study was to reveal how activated sludge communities respond to influent quality and indigenous communities by treating two produced waters from different origins in a batch reactor in succession. The community shift and compositions were investigated using Polymerase Chain Reaction–denaturing gradient gel electrophoresis(PCR–DGGE) and further16 S ribosomal DNA(r DNA) clone library analysis. The abundance of targeted genes for polycyclic aromatic hydrocarbon(PAH) degradation, nah Ac/phn Ac and C12O/C23 O, was tracked to define the metabolic ability of the in situ microbial community by Most Probable Number(MPN) PCR. The biosystem performed almost the same for treatment of both produced waters in terms of removals of chemical oxygen demand(COD) and PAHs. Sludge communities were closely associated with the respective influent bacterial communities(similarity 60%), while one sludge clone library was dominated by the Betaproteobacteria(38%) and Bacteriodetes(30%)and the other was dominated by Gammaproteobacteria(52%). This suggested that different influent and water quality have an effect on sludge community compositions. In addition, the existence of catabolic genes in sludge was consistent with the potential for degradation of PAHs in the treatment of both produced waters.     

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.     

Effect of organic wastes on the plant-microbe remediation for removal of aged PAHs in soils

The effectiveness of in-situ bioremediation of polycyclic aromatic hydrocarbons (PAHs) may be inhibited by low nutrients and organic carbon. To evaluate the effect of organic wastes on the PAHs removal efficiency of a plant-microbe remediation system, contaminated agricultural soils were amended with different dosages of sewage sludge (SS) and cattle manure (CM) in the presence of alfalfa (Medicago sativa L.) and PAHs-degraders (Bacillus sp. and Flavobacterium sp.). The results indicated that the alfalfa mean biomasses varied from 0.56 to 2.23 g/pot in root dry weight and from 1.80 to 4.88 g/pot in shoot dry weight. Low dose amendments, with rates of SS at 0.1% and CM at 1%, had prominent effects on plant growth and soil PAHs degradation. After 60-day incubation, compared with about 5.6% in the control, 25.8% PAHs removal was observed for treatments in the presence of alfalfa and PAHs-degraders; furthermore, when amended with different dosages of SS and CM, the removed PAHs from soils increased by 35.5%-44.9% and 25.5%-42.3%, respectively. In particular, the degradation of high-molecular-weight PAHs was up to 42.4%. Dehydrogenase activities (DH) ranged between 0.41 and 1.83 μupg triphenylformazan/(g dry soil. hr) and the numbers of PAHs-degrading microbes (PDM) ranged from 1.14× 10⁶ to 16.6× 10⁶ most-probable-number/g dry soil. Further investigation of the underlying microbial mechanism revealed that both DH and PDM were stimulated by the addition of organic wastes and significantly correlated with the removal ratio of PAHs. In conclusion, the effect of organic waste application on soil PAHs removal to a great extent is dependent on the interactional effect of nutrients and dissolved organic matter in organic waste and soil microorganisms.     

表层岩溶泉水中多环芳烃污染特征及来源解析

对重庆市南川区4个典型表层岩溶泉进行连续采样观测,利用GC-MS定量分析泉水中16种优控多环芳烃(PAHs)含量,研究了表层岩溶泉水中多环芳烃含量、组成、来源的季节变化以及污染特征.结果表明,4个表层岩溶泉中16种PAHs均有检出.在一年的观测期间,泉水中的ΣPAHs含量变化较大,范围在341~4968 ng·L-1之间,平均值为1772 ng·L-1.7种致癌性PAHs的含量均表现为雨季大于旱季.泉水中PAHs组成以2~3环PAHs为主,其比例均超过了50%.泉水中的PAHs主要来自于煤、生物质以及石油的燃烧,但在2011年6~10月期间,来源于煤、生物质燃烧的PAHs对泉水中PAHs的贡献要更大一些.Ant/(Ant+Phe)和Fla/(Fla+Pyr)比值随季节变化的特点,表明Ant、Phe、Fla和Pyr在表层岩溶带土壤中易于迁移、传输.4个同分异构体比值中,Fla/(Fla+Pyr)比值对源信息的指示更灵敏.与其它地区相比,研究区表层岩溶泉水中的PAHs含量处于较高水平,泉水已受到多环芳烃的污染.     

Degradation of polycyclic aromatic hydrocarbons by Pseudomonas sp. JM2 isolated from active sewage sludge of chemical plant

It is important to screen strains that can decompose polycyclic aromatic hydrocarbons (PAHs) completely and rapidly with good adaptability for bioremediation in a local area. A bacterial strain JM2, which uses phenanthrene as its sole carbon source, was isolated from the active sewage sludge from a chemical plant in Jilin, China and identified as Pseudomonas based on 16S rDNA gene sequence analysis. Although the optimal growth conditions were determined to be pH 6.0 and 37℃, JM2 showed a broad pH and temperature profile. At pH 4.5 and 9.3, JM2 could degrade more than 40% of fluorene and phenanthrene (50 mg/L each) within 4 days. In addition, when the temperature was as low as 4℃, JM2 could degrade up to 24% fluorene and 12% phenanthrene. This showed the potential for JM2 to be applied in bioremediation over winter or in cold regions. Moreover, a nutrient augmentation study showed that adding formate into media could promote PAH degradation, while the supplement of salicylate had an inhibitive effect. Furthermore, in a metabolic pathway study, salicylate, phthalic acid, and 9-fluorenone were detected during the degradation of fluorene or phenanthrene. In conclusion, Pseudomonas sp. JM2 is a high performance strain in the degradation of fluorene and phenanthrene under extreme pH and temperature conditions. It might be useful in the bioremediation of PAHs.     

Response of anaerobes to methyl fluoride, 2-bromoethanesulfonate and hydrogen during acetate degradation

To use the selective inhibition method for quantitative analysis of acetate metabolism in methanogenic systems,the responses of microbial communities and metabolic activities,which were involved in anaerobic degradation of acetate,to the addition of methyl fluoride(CH3F),2-bromoethanesulfonate(BES)and hydrogen were investigated in a thermophilic batch experiment.Both the methanogenic inhibitors,i.e.,CH3F and BES,showed their effectiveness on inhibiting CH4 production,whereas acetate metabolism other than acetoclastic methanogenesis was stimulated by BES,as reflected by the fluctuated acetate concentration.Syntrophic acetate oxidation was thermodynamically blocked by hydrogen(H2),while H2-utilizing reactions as hydrogenotrophic methanogenesis and homoacetogenesis were correspondingly promoted.Results of PCR-DGGE fingerprinting showed that,CH3F did not influence the microbial populations significantly.However,the BES and hydrogen notably altered the bacterial community structures and increased the diversity.BES gradually changed the methanogenic community structure by affecting the existence of different populations to different levels,whilst H2 greatly changed the abundance of different methanogenic populations,and induced growth of new species.     

Effect of nitrite on hydrolysis-acidification, biogas production and microbial community in semi-continuous two-phase anaerobic digestion of sewage sludge

Previous study found that the pre-treatment of sewage sludge with nitrite improves the biogas production during the mono/two-phase anaerobic digestion (AD) using batch biochemical methane potential tests. In this study, the effects of nitrite on hydrolysis-acidification, biogas production, volatile solids destruction and microbial composition in semi-continuous two-phase AD of sewage sludge were investigated. The addition of nitrite promotes sludge organic matter solubilization (+484%) and VFAs production (+98.9%), and causes an increase in the VS degradation rate during the AD process (+8.7%). The comparison of biogas production from the acidogenic and methanogenic reactors with or without the addition of nitrite implies that the nitrite has no significant effect on the overall biogas production of two-phase sludge AD process. High-throughput sequencing analysis shows that the microbial communities of bacteria and archaea in two-phase AD reactors significantly changes after the addition of nitrite. Vulcanibacillus (bacteria) and Candidatus Methanofastidiosum (archaea) become the dominant genera in the acidogenic and methanogenic reactors with the nitrite respectively. These findings provide new insights about using nitrite to promote the organic matter degradation of sewage sludge in a semi-continuous two-phase AD system.     

厦门杏林湾水系表层沉积物中PAHs分析与风险评估

利用ASE-GC-MS(加速溶剂萃取与气相色谱质谱仪联用)方法分析了USEPA(美国环保署)16种优控PAHs在厦门城郊杏林湾水系的19个表层沉积物样点中的含量,并对其组成、来源和风险进行了探讨.结果表明,杏林湾水系表层沉积物中PAHs含量介于413.00~2 748.81 ng·g-1,均值为949.56 ng·g-1;在检测出的13种PAHs中,强致癌性的Bk F和Bghi P检出率高达73.68%,均值分别为69.15 ng·g-1和49.86 ng·g-1;PAHs以2~4环为主,其中2+3环比例均值为61.03%,4环比例均值为23.53%;5和6环均值为15.82%.在所采集的沉积物样品中,中度污染占调查样品数的68.42%,高度污染的占31.58%.利用Ant/(Ant+Phe)和Fla/(Fla+Pyr)比值法和主成分分析方法对杏林湾流域中PAHs其来源进行分析,以及平均沉积物质量基准商(m SQG-Q)法进行生态风险评估,结果表明其PAHs的主要来源为石油源以及石油燃烧源;SQG-Q小于0.50;综合含量、组成特征以及沉积物质量基准商等评价表明,研究区域内靠近杏林工业区一侧的2、3、5和9号采样区以及13号港头采样区表层沉积物中PAHs具有较高的生态安全风险;值得进一步关注.     

降解纤维素产甲烷的四菌复合系

自然环境中通常是微生物群落协同完成纤维素的降解,构建可降解纤维素的混菌体系是认识微生物相互作用的关键.利用富集培养法,结合变性梯度凝胶电泳(DGGE)指纹检测技术以及厌氧滚管技术,建立了一种筛选简单降解纤维素产甲烷复合菌系的方法.利用此方法从青藏高原若尔盖高寒湿地分离到一个由4株菌构成的降解纤维素产甲烷的稳定菌系.结果表明,该复合菌系由纤维素水解菌Clostridium glycolicum、非纤维素水解菌Trichococcus flocculiformis和Parabacteroides merdae、产甲烷古菌Methanobacterium subterraneum等具有不同功能的4种菌株组成,且在这4株菌的共同作用下可将纤维素直接转化为CH4.该简单复合系的获得为今后纤维素转化甲烷复合菌系的代谢控制和遗传改造提供了一个平台.     

象山港海水和沉积物中多环芳烃分布特征和来源研究

本文研究了象山港多环芳烃（PAHs）在沉积物-海水中的分布特征,于2017年1月对港口9个采样点的沉积物和海水中16种优先控制PAHs进行分析,采用同分异构比值法和逸度方法进行来源分析和扩散行为研究。沉积物中PAHs范围为17.51×10–9～84.41×10–9,主要为高环PAHs,处于轻度污染等级。沉积物中多环芳烃主要来自高温燃烧源。表层水体、中层水体和底层水体中PAHs范围分别为41.78～105.72 ng/L、41.51～106.34 ng/L和9.18～145.17 ng/L,主要由低环PAHs组成。表层水体中PAHs主要来源于石油泄漏和石油燃烧。利用逸度系数判断PAHs扩散行为,萘（Nap）、苊（Ace）和芴（Flo）由沉积物向海水释放;苊烯（Acy）、菲（Phe）、蒽（Ant）和芘（Pyr）在沉积物和海水中处于动态平衡;荧蒽（Flu）、苯并[a]蒽（BaA）、?（Chr）和苯并[b]荧蒽（BbF）主要从海水向沉积物扩散并富集。     

西安市地表灰尘中PAHs健康风险特征

为揭示人群暴露地表灰尘中PAHs的健康风险,在西安市采集58个地表灰尘样品,分析其中16种优控PAHs质量分数,并根据美国能源部风险评估信息系统中的暴露方法对其健康风险进行评价.结果表明:西安市地表灰尘中w(∑₁₆PAHs)(16种PAHs总量)范围为5.04～47.74 mg/kg,平均值为13.85 mg/kg.人群暴露地表灰尘中PAHs的主要途径是经口摄入,并且不同途径下儿童的暴露剂量均高于成人.地表灰尘中PAHs对人群没有明显的非致癌健康危害,但对儿童的非致癌危害高于成人,其中Nap、Phe、Fla、Pyr和BghiP对人群的非致癌风险明显高于Acy、Ace、Flu和Ant.7种致癌PAHs的致癌风险大小顺序为BaP>DBA>BbF>InP>BaA>BkF>Chy,致癌风险总和为2.51×10-5,其中BaA、BbF、BaP、InP和DBA致癌风险在1.07×10-6～9.56×10-6之间.研究显示,西安市地表灰尘中16种PAHs对人群的健康危害相对较低.      

焦化厂不同污染源作用下土壤PAHs污染特征

根据生产工序的不同将焦化场地划分为堆煤区、炼焦区、化产区,共采集40组土壤样品,分析各类污染源作用下场地PAHs污染程度、分布、影响途径及组成特征等.结果表明,场地处于严重污染水平且BaP是健康风险首要关注污染物.按ΣPAHs含量中位数排序,化产区（1733.87mg/kg）>炼焦区（32.86mg/kg）>堆煤区（21.21mg/kg）.对应污染途径依次为化工副产品的泄漏及填埋、烟粉尘大气沉降、煤粉（渣）降雨淋滤.异构体比值法判定的污染源不能明显区分各工序的土壤污染特点且存在偏差,利用ω（低环PAHs）/ω（高环PAHs）比值法进行排序,化产区深层（7.39）>化产区表层（1.33）>堆煤区（1.06）>炼焦区（0.39）,PAHs组成特征受污染源自身特性及外环境作用共同所致.4~5环PAHs是该焦化场地的特征污染物,化产区、堆煤区土壤中Nap、Phe占ΣPAHs比重较高,而炼焦区以BbF、Fla、Chry为主要组分.     

Microbial community structures in an integrated two-phase anaerobic bioreactor fed by fruit vegetable wastes and wheat straw

The microbial community structures in an integrated two-phase anaerobic reactor（ITPAR）were investigated by 16 S r DNA clone library technology. The 75 L reactor was designed with a 25 L rotating acidogenic unit at the top and a 50 L conventional upflow methanogenic unit at the bottom, with a recirculation connected to the two units. The reactor had been operated for 21 stages to co-digest fruit/vegetable wastes and wheat straw, which showed a very good biogas production and decomposition of cellulosic materials. The results showed that many kinds of cellulose and glycan decomposition bacteria related with Bacteroidales,Clostridiales and Syntrophobacterales were dominated in the reactor, with more bacteria community diversities in the acidogenic unit. The methanogens were mostly related with Methanosaeta, Methanosarcina, Methanoculleus, Methanospirillum and Methanobacterium; the predominating genus Methanosaeta, accounting for 40.5%, 54.2%, 73.6% and 78.7% in four samples from top to bottom, indicated a major methanogenesis pathway by acetoclastic methanogenesis in the methanogenic unit. The beta diversity indexes illustrated a more similar distribution of bacterial communities than that of methanogens between acidogenic unit and methanogenic unit. The differentiation of methanogenic community composition in two phases, as well as pH values and volatile fatty acid（VFA） concentrations confirmed the phase separation of the ITPAR. Overall, the results of this study demonstrated that the special designing of ITPAR maintained a sufficient number of methanogens, more diverse communities and stronger syntrophic associations among microorganisms, which made two phase anaerobic digestion of cellulosic materials more efficient.     

焦化厂不同污染源作用下土壤PAHs污染特征

根据生产工序的不同将焦化场地划分为堆煤区、炼焦区、化产区,共采集40组土壤样品,分析各类污染源作用下场地PAHs污染程度、分布、影响途径及组成特征等.结果表明,场地处于严重污染水平且BaP是健康风险首要关注污染物.按ΣPAHs含量中位数排序,化产区（1733.87mg/kg）>炼焦区（32.86mg/kg）>堆煤区（21.21mg/kg）.对应污染途径依次为化工副产品的泄漏及填埋、烟粉尘大气沉降、煤粉（渣）降雨淋滤.异构体比值法判定的污染源不能明显区分各工序的土壤污染特点且存在偏差,利用ω（低环PAHs）/ω（高环PAHs）比值法进行排序,化产区深层（7.39）>化产区表层（1.33）>堆煤区（1.06）>炼焦区（0.39）,PAHs组成特征受污染源自身特性及外环境作用共同所致.4~5环PAHs是该焦化场地的特征污染物,化产区、堆煤区土壤中Nap、Phe占ΣPAHs比重较高,而炼焦区以BbF、Fla、Chry为主要组分.     

低环多环芳烃异位热脱附行为分析

随着我国工业和城镇的快速发展,化石能源的大量使用使得环境中多环芳烃(PAHs)残留量不断增加,给环境和人体健康带来严重危害.以2～4环PAHs为研究对象,探讨了热脱附温度、时间以及PAHs自身结构对热脱附效果的影响,简要分析了热脱附处理对土壤性质的影响,评估了管式炉热脱附的能耗.结果表明:低环PAHs的脱附率随着脱附温...