Porphyrobacter sp.D22F的降解特性及其在石油降解菌群中的生态位

为揭示多环芳烃(Polycyclic aromatic hydrocarbons,PAHs)降解微生物资源的多样性和石油降解菌群的优势菌,研究了从南海沉积物中分离得到的一株PAHs降解菌D22F的降解特性及其在石油降解菌群D22-1中的生态位.对菌株D22F进行16S rRNA基因同源性分析及透射电镜观察以初步确定其种属,通过培养法、气相色谱质谱联用(GC-MS)测定其多环芳烃降解范围和降解率,通过简并引物PCR扩增其PAHs双加氧酶大亚基基因片段并进行系统发育分析,采用变性梯度凝胶电泳(DGGE)监测石油降解菌群中的优势菌.结果表明,与菌株D22F的16S rRNA基因相似度最高的模式株为产卟啉杆菌属Porphyrobacter tepidarius DSM 10594T(AF465839;98.55%).该菌株能降解萘、甲基萘、苊、硫芴、菲、蒽等;对初始浓度为0.2 g/L菲10 d后的降解率可达90%以上.从其基因组DNA中克隆到的PAHs起始双加氧酶大亚基基因phnAc与Novosphingobium aromaticivorans DSM 12444中的质粒pNL1(CP000676)上的bphA1f基因相似度最高,达到99.41%.DGGE谱图显示,菌株D22F是石油降解菌群中的3种优势菌之一,在传代菌群中可稳定存在.Porphyrobacter sp.D22F为产卟啉杆菌属(Porphyrobacter)中首株以低分子量PAHs为唯一碳源和能源的菌株,是石油降解菌群的优势菌.     

Passive sampling of PAHs in indoor air in Nepal

PAHs were sampled in ten homes in the Makwanpur region, Nepal. SPMDs and moss bags (Sphagnum girgensohnii) were used as passive samplers. Soot particles on the SPMD surfaces were also analyzed for PAHs. The overall PAH concentrations in SPMDs were significantly higher than those in moss bags. Total PAH mean concentrations of ten houses were 535μg/g lipid for SPMDs and 7.2 μg/g moss (dw) for moss bags. Ratios of phenanthrene/anthracene in indoor SPMDs and particulate matter varied from 2.9 to 3.5 and ratios of fluoranthene/pyrene varied from 1.1 to 1.4. The values for moss bags were respectively 1.7–3.6 and 0.8–2.4. These ratios indicate that the PAHs are from combustion origin. The PAH concentrations in ambient air were estimated as B(a)P TEQs and they were 17–64 times higher than acceptable limit for Finnish community air. Based on PAH levels in the gas phase (SPMD) we may expect PAHs to have an impact on respiratory disease prevalence in Nepalese villages. Both of the sampling methods were feasible in the difficult conditions under which the study was performed.     

Biodegradation and phytotoxicity assessment of phenanthrene by biosurfactant-producing Bacillus pumilus 1529 bacteria

ABSTRACT

Phenanthrene is a toxic and mutagenic pollutant that can cause severe environmental and human health issues. The bioremediation of these polyaromatic hydrocarbons (PAHs) is possible with a biosurfactant by enhancing hydrophobicity. In this study, the production of a biosurfactant by Bacillus pumilus 1529 and its effects on the phenanthrene biodegradation pathway were examined. Biosurfactant production was determined using hemolytic activity, emulsification index, and surface tension. For phenanthrene metabolite detection, samples at 0, 7, 14, and 21 incubation days were analysed by gas chromatography-mass (GC-mass) spectrometry. The results showed that Bacillus pumilus 1529 can reduce surface tension to 22.83?±?1.1?mN?m^?1. Furthermore, the GC-mass spectrometry analysis showed that 1-hydroxy-2-naphthoic acid, benzaldehyde, o-phthalic acid, and phenylacetic acid were notable phenanthrene metabolites produced during phenanthrene biodegradation. Biodegraded phenanthrene and its metabolites have a less toxic effect on the germination of safflower seeds than non-biodegraded phenanthrene. The IC50 of phenanthrene on seed germination after biodegradation was increased to approximately 113?mg?L^?1. In general, biodegradation aided by biosurfactant producing bacteria contributed to turning the toxic phenanthrene into less harmful metabolites with lower phytotoxicity effects, indicating that its application in the bioremediation of PAHs is promising.     

Phytotoxicity assessment of phenanthrene and pyrene in soil using two barley genotypes

This study investigated the toxicity extent of phenanthrene and pyrene to two cultivars (CM-72 and Gairdner) of barley (Hordeum vulgare). Germination of barley seeds was evaluated in 69-d aged soil, separately spiked with phenanthrene at extractable concentrations of 0.95, 6.3, 59, and 300 mg kg^?1 (dry soil) and pyrene at 1.0, 9.0, 73, and 400 mg kg^?1 (dry soil). Although germination was not inhibited, significant (P < 0.05) reduction in root and shoot length occurred at concentrations of phenanthrene ≥6.3 mg kg^?1 and pyrene ≥9.0 mg kg^?1 after both 72 and 240 h. Fresh and dry biomass of both cultivars reduced with increasing concentrations of both polycyclic aromatic hydrocarbons (PAHs). Barley cultivar CM-72 was more sensitive than Gairdner, and it can be considered suitable for toxicity assessment of PAH-contaminated soils.     

多环芳烃降解菌ZL5分离鉴定及其降解质粒

通过选择性富集培养，从辽河油田石油污染土壤中分离到一株多环芳烃(PAHs)降解菌ZL5．它能以菲和芘为唯一碳源生长，但是不能利用萘．16S rDNA核苷酸序列分析结果表明，ZL5属于变形细菌α亚类中的鞘氨醇单胞菌属．该菌株含有一个大小约为60kb的质粒．丝裂霉素C消除实验表明，随着质粒的丢失，菌株利用菲和芘的能力也丧失．用电转化和氯化铷转化法分别将菌株ZL5的质粒导人大肠杆菌JM109和DH5α中，随着质粒的获得，这些转化子获得了降解菲和芘的能力．本研究结果表明，鞘氨醇单胞菌ZL5降解PAHs的功能和质粒有关。     

Multi-metal resistance and plant growth promotion potential of a wastewater bacterium Pseudomonas aeruginosa and its synergistic benefits

Water and soil pollution by toxic heavy metals (HMs) is increasing globally because of increase in population, industrialization and urbanization. It is a burning problem for the public, scientists, academicians and politicians how to tackle the toxic contaminants which jeopardize the environment. One possible solution for pollution abatement is a bioremediation-effective and innovative technology that uses biological systems for treatment of contaminants. Many bacteria synthesize indole-3-acetic acid (IAA) which is a product of l-tryptophan metabolism and belongs to the auxin class of plant growth-promoting hormone. The present study aimed at assessing the resistance pattern of wastewater bacteria against multiple HMs and plant growth promotion activity associated with IAA. A Gram-negative bacterial strain Pseudomonas aeruginosa KUJM was isolated from Kalyani Sewage Treatment Plant. This strain showed the potential to tolerate multiple contaminations such as As(III) (50 mM), As(V) (800 mM), Cd (8 mM), Co (18 mM), Cu (7 mM), Cr (2.5 mM), Ni (3 mM) and Zn (14 mM). The capability of IAA production at different tryptophan concentration (1, 2, 5 and 10 mg mL⁻¹) was determined, and seed germination-enhancing potential was also estimated on lentil (Lens culinaris). Such type of HM-resistant, IAA-producing and seed germination-enhancing P. aeruginosa KUJM offer great promise as inoculants to promote plant growth in the presence of toxic HMs, as well as plant inoculant systems useful for phytoremediation of polluted soils. Hence, P. aeruginosa KUJM finds significant applications in HM-contaminated poor agricultural field as well as in bioremediation of HM-contaminated wastewater system.

     

Mikrobieller PAK-Abbau

The biodegradation of the five ring PAH benzo[a]pyrene (BaP) is assumed to be limited by the low water solubility of this compound. A mixed culture of microorganisms — isolated from a PAH-contaminated soil — was able to degrade¹⁴C labelled BaP in mineral medium by cometabolism with phenanthrene, fluoranthene, anthracene and pyrene as sources of carbon and energy. The mineralisation of these compounds to low levels resulted in an inhibition of the degradation of BaP. After the new addition of the four PAH compounds to the culture medium the mineralisation of BaP started again. A non-ionic surfactant of the alkylpolyglycoside type (Plantacare 2000 UP) increased the concentration of BaP in the culture medium because of solubilization. At high Plantacare concentrations, the degradation of BaP was completely inhibited above the critical micelle concentration (cmc). The degradation of the three and four ring PAHs was also inhibited. If the surfactant was metabolised to concentrations below the cmc, an increase of mineralisation of BaP could occur up to 24% in 384 days.     

Occurrence of PCBs and PAHs in an urban soil of Madrid (Spain)

Polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs) were analysed in surface soil samples collected in 2001–2002 at an open urban area of Madrid. In order to obtain representative samples, three collection points at the site were chosen. The most abundant PAHs and PCBs were phenanthrene, fluorene, pyrene, chrysene and benzo[a]pyrene and hexa- and heptachlorinated PCBs, respectively. The sum of selected PAHs (13 compounds) and PCBs (15 congeners) averaged 1 and 0.1?µg?g^?1, respectively. PCB levels in winter were 2–10 times higher than summer ones, while seasonal variation for most of PAHs were not observed. Good correlations among all PCBs were found. Five PAHs were also well correlated.     

Assessment of PAHs in water and fish tissues from Great Bitter and El Temsah lakes,Suez Canal,as chemical markers of pollution sources

Sea water and fish tissue samples were collected from nine sampling stations from the Great Bitter and El Temsah lakes in the Suez Canal and analysed for polycyclic aromatic hydrocarbon (PAH). The compositions of PAH determined in the dissolved fraction of sea water were measured in order to use them as chemical markers for identifying different sources of PAH pollution in this region. PAHs determined in fish tissues were measured for comparison with human health standards as consumption. The total mean PAHs concentrations in the sea water samples ranged from 0.28 to 39.57 μg l^?1 with an overall mean of 10.78 and 12.38 μg l^?1 for El Temsah and Bitter Lakes water, respectively. Total PAHs fractions recorded in muscle tissues of all different Osteicthyes fishes collected from Great Bitter lakes ranged from 5.8 to 218.5 μg g^?1 with an overall mean of 57.98 μg g^?1 during all seasons. However, they ranged from 68 to 623 μg g^?1 with an overall mean of 87.69 μg g^?1 recorded in El Temsah lake during four seasons (2003–2004). Benzo(a)pyrene was the most dominant PAHs found in the sea water samples from both lakes with an average concentration of 3.8 μ g l^?1. Dibenzo(a,h)anthracene (DBA) was the most dominant PAHs recorded in fish samples. A maximum of 533 μg g^?1 of DBA was recorded in Dahbana sp. collected from Bitter lakes during January 2004. However, a maximum of 68.7 μ g g^?1 was recorded in Liza carinata species collected from El Temsah lake during July, 2004. The simultaneous occurrence of isomer ratios PHE/ANT<10 for all stations indicated that the major PAH input to water was from combustion of fossil fuel (pyrolytic source). The average ratios were 1.21 and 12.9 during winter (January 2004) and 4.3 and 8.63 during spring (April 2004) for all water samples of Great Bitter lakes and El Temsah lake, respectively. In addition, the present data demonstrate that PAHs from fossil fuel sources (MW<178) were the least significant source of PAHs in this region.     

Performance evaluation of isoproturon-degrading indigenous bacterial isolates in soil microcosm

Isoproturon (IPU)-degrading soil bacteria were isolated from herbicide-applied wheat fields. These isolates were identified using cultural, morphological, biochemical and 16S rRNA sequencing methods. 16S rRNA sequences of both the bacterial isolates were compared with NCBI GenBank data base and identified as Bacillus pumilus and Pseudoxanthomonas sp. A soil microcosm study was carried out for 40 days in six different treatments. Experimental results revealed maximum 95.98% IPU degradation in treatment 6 where bacterial consortia were augmented in natural soil, followed by 91.53% in treatment 5 enriched with organic manure as an additional carbon source. However, only 14.03% IPU was degraded in treatment 1 (control) after 40 days. In treatments (2–4), 75.59%, 70.92% and 77.32% IPU degradation was recorded, respectively. IPU degradation in all the treatments varied significantly over the control. 4-Isopropylaniline was detected as IPU degradation by-product in the medium. The study confirmed that B. pumilus and Pseudoxanthomonas sp. performed effectively in soil microcosms and could be employed profitably for field-scale bioremediation experiments.     

Concentration of some polycyclic aromatic hydrocarbons in rain,soil and ground water around Ismailia,Egypt

Residues of acenaphthylene, fluorene, anthracene, pyrene, chrysene, benzo(b)fluoranthene and benzo(k)fluoranthene were monitored in rain, soil and groundwater around Ismailia, in northeast Egypt. Residues detected in rain water in 1995 and 1996 were mainly of relatively low molecular weight. Both acenaphthylene and fluorene were detected in rain for the two consecutive years. Top soil has shown a wider spectrum and higher concentrations of (PAHs) than those detected in deep soil, rain and ground water. Only three compounds, acenaphthylene, fluorene and anthracene were detected in samples collected at 50 cm depth. While no traces of PAHs were detected at 1 m depth detectable concentrations of fluorene and anthracene were monitored in groundwater.     

Degradation of benzyldimethyl hexadecylammonium chloride by Bacillus niabensis and Thalassospira sp. isolated from marine sediments

Following enrichment in its presence, two strains of bacteria, isolated from marine sediments, were shown to degrade the quaternary ammonium surfactant benzyldimethyl hexadecylammonium chloride (BDHAC) in a minimal salts medium. The bacteria identified by 16S ribosomal deoxyribonucleic acid sequencing were shown to belong to several genera and determined to be Bacillus niabensis and Thalassospira sp. Initial investigations demonstrated that the bacteria were capable of degrading BDHAC when it was present at concentrations in the range 2–4 mg mL^?1. In media containing BDHAC, up to 90% was degraded within 7 days, but limited growth of the strain was observed at 2 and 4 mg mL^?1 BDHAC. Preliminary analysis of samples after degradation experiment by electrospray ionization mass spectrometry/mass spectrometry produced a peak with a parent–daughter ion transition of 136 → 91, corresponding to N,N-dimethylbenzylamine. The presence of this potential metabolite suggests the cleavage of the C-alkyl-N bond as a step in BDHAC catabolism.     

一株菲降解菌的特性及相关降解基因的克隆

从石油污染土壤中分离到一株菲降解菌2F5-2.根据该菌株生理生化特征和16S rDNA序列相似性分析,将其初步鉴定为鞘氨醇杆菌属(Sphingobium sp.).该菌株在10 h内对100 mg/L的菲的降解率为100%.降解菲的最适温度为30℃,最适pH为7.对降解途径的初步研究显示,该菌株通过水杨酸途径降解菲.克隆了编码芳香烃双加氧酶α亚基的基因phdA,它与菌株Sphingomonas sp.P2、Sphingobium yanoikuyae B1、Sphingomonas sp.ZP1中phdA的同源性分别为97.9%、98%和100%,表明该基因具有保守性.图6参16     

Mutagenicity of polyaromatic hydrocarbons by chemical models for cytochrome P450 in Ames assay

DNA damage is an important step in carcinogenesis. The Ames assay is a short-term screening of carcinogens that induce DNA damage. Most carcinogens require enzymatic activation through oxidation by cytochrome P450 (CYP450) in the presence of S9 mix. A combination of iron (Fe)(III) porphyrin and an oxidant is also able to oxidize compounds as an alternative metabolic pathway to CYP450. Previously it was reported that a chemical model containing a water-soluble 5,10,15,20-tetrakis(1-methylpyridinium4-yl)porphyrinatoiron(III) chloride (4-MPy) and tert-butyl hydroperoxide (t-BuOOH) activated aromatic amines and amides. In this study, a chemical model composed of an Fe porphyrin, water-insoluble 5,10,15,20-tetrakis(pentafluorophenyl)porphyrinatoiron(III) chloride (F₅P) or water-soluble 4-MPy was optimized with an oxidant – t-BuOOH, magnesium monoperoxyphthalate (MPPT), or iodosylbenzene (PhIO). Subsequently the mutagenicity of benzo[a]pyrene (B[a]P) and chrysene in Salmonella typhimurium TA strains was compared. B[a]P was activated by a combination of F₅P or 4-MPy plus MPPT or PhIO in S. typhimurium TA1538. The B[a]P-induced mutagenicity with F₅P plus oxidant was higher than 4-MPy plus oxidant. Mutagenicity of chrysene, a tetracyclic aromatic hydrocarbon, was not detected in the presence of F₅P/PhIO in S. typhimurium TA98, but was activated in the presence of F₅P/MPPT. The F₅P/MPPT activated other polyaromatic hydrocarbons (PAH) in the S. typhimurium TA98 assay including dibenz[a,c]anthracene, dibenz[a,h]anthracene, 3-methylcholanthrene, and benzo[a]anthracene. The results indicated that the F₅P/MPPT was the most efficient model for detecting PAH-induced mutagenicity in the Ames assay.     

Determination of persistent organic pollutants in sediment and fish of the western coast of Alexandria,Egypt

Persistent organic pollutants (POPs) were recorded in sediment and fish samples collected from the western coast of Alexandria. Total hydrocarbons (aliphatic+PAHs ) in sediment ranged from 683.8 to 34670.1 ng g ^?1 with an average of 9286.9 ng g ^?1. The sum of C16–C34 of aliphatic fractions was<4000?ng g;^?1, indicating the presence of a fresh petroleum source. For all sediments, the anthracene/phenanthrene ratio was>0.1, suggesting the dominance of a pyrolytic source. Total aliphatics in different fish species ranged from 253 to 11?132 ng g;^?1, while total PAHs ranged from 3862 to 35?746 ng g;^?1 wet weight. Benzo[a]pyrene was the most dominant PAH fraction ranged from 1902.7 to 32 905.5 with an average of 9464.5?ng g;^?1 wet weight in all fish species. Concentrations of polychlorinated biphenyls (PCBs) ranged from 0.79 to 64.9?ng g;^?1 with an average 12.14?ng g;^?1 wet weight. The concentrations of organochlorines in fish species (Euthynnus alleferatus, Scomberomorus commerson, Sphyraena Sphyraena, Diplodus vulgaris, and Alepes djedaba) decreased following the order: PCBs>DDTs>HCHs>total cyclodienes. Concentrations of DDTs in fish tissues ranged from 4.89 to 36.37 ng g^?1 with an average of 16.4?ng g;^?1 wet weight. The concentrations of total HCHs ranged from 0.3 to 65.7?ng g;^?1 with an average of 16.35?ng g;^?1. The present study indicates: (1) fresh petroleum input where Pr/Ph>1; (2) PAHs in sediment<4000 ng g ^?1; (3) BaP concentration exceeded the permissible levels in Alepes djedaba species; (4) DDTs in sediment were below the effective range low level; (5) PCBs>effective range low and相似文献   

Assessing the bioavailability of phenanthrene to soil microorganisms using the Tenax extraction method

The Tenax TA extraction technique followed by gas chromatography mass spectrometry (GC-MS) determinations was used to assess the actually bioavailable fraction of phenanthrene (ABF-Phe) in three different soils freshly contaminated with this compound at levels of 10, 100, and 1,000 mg kg⁻¹. The results were related to the ecotoxic effect of phenanthrene on soil nitrifying bacteria. Nitrification potential measurements were applied as an ecotoxicity end point. A strong positive linear relationship (r ² = 0.95) was obtained between the content of the actually bioavailable phenanthrene fraction and the inhibition of nitrifying bacteria activity.     

菲和镉单一及复合污染条件下在毛蚶体内的富集动力学研究

采用室内半静态双箱动力学模型实验,研究了菲和镉单一及复合污染条件下在毛蚶(Anadara subcrenata)体内的生物富集,通过对富集与释放过程中毛蚶体内菲和Cd的富集量进行非线性曲线拟合,获得了菲和Cd单一及复合污染条件下在毛蚶体内的吸收速率常数k1、释放速率常数k2、生物富集因子BCF、生物半衰期B1/2和平衡状态下最大富集量CA max等动力学参数。实验结果表明,菲和Cd在实验前期富集速率较高,8 d以后富集速率减缓,释放阶段与富集阶段相似。毛蚶对菲的BCF值为37.80,远大于Cd的BCF值13.12,且生物半衰期时间更长,菲更容易在生物体内富集。菲和Cd联合暴露条件下,在毛蚶体内的CA max和BCF值均大于单一作用,说明二者同时暴露时,毛蚶对菲和镉的吸收富集均有所增强。实验模型拟合度较好,输出值和实测值之间无显著性差异,拟合方程和拟合参数可信。     

PAHs in organic film on glass window surfaces from central Shanghai,China: distribution,sources and risk assessment

Polycyclic aromatic hydrocarbons (PAHs) concentrations were analysed in the organic film on the glass surfaces of different functional areas in central Shanghai. Concentration levels of total PAHs in the organic film ranged from 1,348.5 to 4,007.9 ng m^?2. The concentration of PAHs was lowest in parks and green spaces (1,348.5 ng m^?2) and highest in traffic zones (4,007.9 ng m^?2). A concentration gradient of total PAHs was observed as follows: traffic zones > commercial areas > cultural and educational areas > parks and green spaces. The distribution of PAHs was characterised by 3–4 ring PAHs in the study areas. The most abundant PAHs were phenanthrene (20.5 %), fluorene (16.7 %), pyrene (12.4 %) and chrysene (Chry) (11.2 %). The mass of the bulk film was composed of organic and inorganic compounds and ranged from 246 to 1,288 mg m^?2. The bulk film thickness varied from 144 to 757 nm in the different functional areas. The ratios of An/178 and Fl/202 and principal component analysis suggested that PAHs came mainly from the mixed sources of fossil fuel, coal and incomplete combustion of biomass. Benzo[a]anthracene (BaA)/Chry is not suitable for use as a tracer for the transmission process of PAHs because of the rapid depletion of BaA in the organic film by photooxidation during daylight hours. The concentration of benzo[a]pyrene equivalent (BaPeq) varied from 21 to 701 ng g^?1, and the major carcinogenic contributors of the 16 PAHs were BaP, DahA, B[b/k]F and InP, accounting for 83 % of BaPeq.     

菲胁迫下湿生植物美人蕉(Canna indica)对斜生栅藻(Scenedesmus obliquus)生长的影响

多环芳烃有机污染物的环境问题日益严重,亟待需求从群落及生态系统的角度进行毒理学复合危害效应的研究。本文采用以美人蕉(Canna indica)种植根区水培养斜生栅藻(Scenedesmus obliquus)的方式,观察菲胁迫下其生长的变化。结果表明,美人蕉种植根区水对斜生栅藻生长具有化感效应,且表现为"低促高抑"的现象。而在菲胁迫下,种植根区水比例为15%和30%时随着菲浓度的增加,生长促进作用增强,且在菲浓度为1 mg·L-1出现最高值;在100%种植根区水时随着菲浓度的增加生长抑制作用增强。说明,菲胁迫使美人蕉种植根区水对藻类的化感效应增强。进行逻辑斯谛生长拟合发现,30%、1 mg·L-1处理组种群内禀增长率最高,斜生栅藻种群出现暴发增长的生态风险最大。     

Biomarkers indicate mixture toxicities of fluorene and phenanthrene with endosulfan toward earthworm (<Emphasis Type="Italic">Eisenia fetida</Emphasis>)

α-Endosulfan and some polycyclic aromatic compounds (PAHs) are persistent in the environment and can reach crop products via contaminated agricultural soils. They may even be present as mixtures in the soil and induce mixture toxicity in soil organisms such as earthworms. In this study, the combined toxicities of PAHs with α-endosulfan were determined in Eisenia fetida adults using an artificial soil system. α-Endosulfan and five PAHs were tested for their acute toxicity toward E. fetida in artificial soils. Only α-endosulfan, fluorene, and phenanthrene showed acute toxicities, with LC₅₀ values of 9.7, 133.2, and 86.2 mg kg^?1, respectively. A mixture toxicity assay was conducted using α-endosulfan at LC₁₀ and fluorene or phenanthrene at LC₅₀ in the artificial soils. Upon exposure to the mixture of fluorene and α-endosulfan, earthworms were killed in increasing numbers owing to their synergistic effects, while no other mixture showed any additional toxicity toward the earthworms. Along with the acute toxicity results, the biochemical and molecular changes in the fluorene- and phenanthrene-treated earthworms with or without α-endosulfan treatment demonstrated that enhancement of glutathione S-transferase activity was dependent on the addition of PAH chemicals, and the HSP70 gene expression increased with the addition of α-endosulfan. Taken together, these findings contribute toward understanding the adverse effects of pollutants when present separately or in combination with other types of chemicals.