真菌对污染水稻土中苯并[a]芘共代谢降解研究

在恒温和恒定转速培养条件下,模拟生物泥浆反应器法,选择从石油污染土壤中分离出来的青霉菌、黑曲霉、白腐真菌等3种真菌,在添加不同浓度菲和邻苯二甲酸作为共存底物情况下,研究其对水稻土中苯并[a]芘（B[a]P）的共代谢降解。结果表明,未灭菌土壤对B[a]P有降解能力。当土壤中添加菲时,提高了B[a]P在土壤中的降解率,100 mg kg-1浓度菲处理的降解率显著高于200 mg kg-1浓度菲处理,邻苯二甲酸对B[a]P降解影响不大。灭菌土壤中的B[a]P几乎没有降解。添加菲及邻苯二甲酸均促进了青霉菌对B[a]P的降解,其中菲浓度为100 mg kg-1处理效果最显著。与灭菌土壤相比,接种黑曲霉提高了B[a]P的降解率,但添加菲与邻苯二甲酸却均抑制了黑曲霉菌对B[a]P的降解。白腐真菌能有效地降解B[a]P,但高浓度菲抑制了白腐真菌对B[a]P的降解,同时邻苯二甲酸对促进白腐真菌降解B[a]P的效果不明显。     

镉和苯并[a]芘胁迫对小麦幼苗生理指标的影响

采用水培方法研究了镉（Cd）和苯并[a1芘（BaP）污染对小麦幼苗生理指标的影响。营养液中镉和苯并[a]芘的浓度分别为10mg·L^-0和54mg·L^-0。结果表明：与对照组相比,各胁迫组小麦生物量下降,株高降低,复合胁迫组地上及地下干重分别下降13．1％和49．8％,株高增长最低;各组超氧化物歧化酶（SOD）活性呈现升高-降低-升高趋势,且低于对照,过氧化物酶（POD）和过氧化氢酶（cAT）活性先升高再降低,复合胁迫组POD活性最高,CAT活性最低。镉、苯并[a]芘及其复合胁迫下,小麦幼苗的生长受到了一定抑制,且以复合胁迫最为显著。镉、苯并[a]芘联合作用的结果表现为协同促进作用。     

土壤pH、水分及温度对长期污染土壤中苯并[a]芘动态变化的影响初探

以受多环芳烃长期污染农田土壤中苯并[a]芘(B[a]P)为研究对象,采用室内模拟试验研究了pH、水分和温度对土壤中B[a]P的动态变化的影响。结果表明,供试污染土壤中B[a]P能被土著微生物快速去除,其消减过程受到pH、水分和温度的影响。初始pH为4.6、5.9和7.3的泥浆中B[a]P在7天内的去除率分别为43.8%、37.8%和14.0%。在土壤水分为22%、43%、65%和87%田间持水量,温度为28℃条件下培养80天后土壤中B[a]P的去除率分别为42.5%、96.6%、96.3%和34.3%。在土壤pH为4.5,土壤水分为60%±5%田间持水量,温度为25℃和40℃条件下土壤中B[a]P的半衰期分别为33天和16天。可见,pH、水分和温度等环境条件对土壤中B[a]P的消减过程具有明显的影响,可通过人为调控强化B[a]P污染土壤的生物修复。     

苯并[a]芘污染土壤的丛枝菌根真菌强化植物修复作用研究

研究了种植紫花苜蓿 (MedicagosativaL )在接种和不接种菌根真菌 (GlomuscaledoniumL )情况下对土壤中苯并 [a]芘 (B[a]P)的降解动态。历经 90天的温室盆栽试验表明 ,较高浓度 (10 0mgkg-1)B[a]P能降低菌根真菌对植物根的侵染率。种植紫花苜蓿和接种菌根真菌能促进土壤中可提取态B[a]P的降解 ,在接种情况下 ,有植物时对三种浓度 (1mgkg-1,10mgkg-1,10 0mgkg-1)B[a]P的降解率分别达 86 2 %、86 6 %、5 7 0 % ;而没有植物时B[a]P的降解率为 5 3 5 %、5 3 0 %、33 0 %。不接菌根真菌时的降解率比接菌根真菌的低得多 ,不接种菌根真菌时 ,有植物的B[a]P降解率分别达 75 9%、77 7%、5 3 4 % ;而不种植物的降解率分别为 5 4 9%、5 2 6 %、34 1% ,低、中浓度 (1mgkg-1,10mgkg-1)两处理的降解率明显地高于高浓度处理(p <0 0 5 )。B[a]P添加对土壤中多酚氧化酶活性有较大的影响 ,特别是高浓度B[a]P处理土壤的酶活性明显地低于其它三个处理 ,接种菌根真菌能够提高土壤中的酶活性 ,从而促进了土壤中B[a]P的降解。     

噬氨副球菌HPD-2固体发酵条件优化及其对PAHs污染土壤的修复效果

选用有机肥A、有机肥B和紫花苜蓿粉3种有机物料作为菌剂载体,探讨了固体发酵过程中物料量、接种量、固水比和发酵时间对噬氨副球菌HPD-2生长的影响,以及固体发酵菌剂对PAHs污染土壤的修复效果。结果表明,3种发酵载体中,以有机肥A的效果最好。噬氨副球菌HPD-2以有机肥A为载体的固体发酵最佳条件为物料量20 g、接种量5%、固水比1:1、发酵时间 144 h。菌剂施入土壤28天后,土壤中PAHs总含量由初始的9.96 mg/kg 下降到7.64 mg/kg,其去除率达22.8%。不同环数PAHs的去除率高低顺序为3环＞5环＞6环和4环,分别下降了35.1%、27.0%、20.7%、20.4%。该菌剂对PAHs污染土壤有一定的修复效果。     

铅和苯并[a]芘混合污染酸性土壤上黑麦草生长及对污染物的吸取作用

设置铅(Pb)处理浓度为0、5001、0002、000 mg kg-1(烘干土计),苯并[a]芘(B[a]P)处理浓度为0、12.5、255、0 mg kg-1(烘干土计),完全组合设计,对每一处理通过种植和不种植黑麦草盆栽试验,研究了B[a]P和Pb混合污染酸性红砂土上黑麦草(Lolium perenneL.)的生长及对污染物积累的吸取作用。试验表明,铅是抑制黑麦草株高和产量的主要因素,在低Pb处理浓度下,B[a]P对黑麦草生长有一定的促进作用。在试验61 d内,黑麦草吸收的铅占土壤铅添加量的4.7%,黑麦草吸收的B[a]P占土壤B[a]P添加量的0.023%;种植黑麦草的土壤B[a]P残留率平均为42.6%;而未种植黑麦草的土壤B[a]P平均残留率为50.9%。该研究结果表明,当Pb-B[a]P混合污染土壤时,在一定的浓度范围内黑麦草能吸收土壤中的Pb和B[a]P,黑麦草对Pb-B[a]P混合污染土壤有一定的吸取修复作用。     

铜-苯并[a]芘复合污染体系中铜的微生物吸附特性

研究了铜-苯并[a]芘（BaP）复合污染体系中,嗜麦芽窄食单胞菌对铜的微生物吸附特性。实验结果表明,该菌可以选择性地吸附Cu^2＋,并把NO3^-还原为NO2^-,pH、投菌量、处理时间和铜浓度等因素及BaP均会对铜的生物吸附产生显著的影响,BaP和这4个因素的相互作用对铜的生物吸附也达到了显著的水平。投菌量会对铜吸附过程中F-、Cl^-、NO2^-、NO3^-、PO34^-和SO24-等离子的释放产生显著的影响,BaP则会显著的影响F^-、NO2^-、NO3^-和PO34^-的浓度水平。当BaP的浓度为0、0.1、1.0、10.0 mg.L^-1时,2.5 g.L^-1菌体对pH为6.0、浓度为2 mg.L^-1的铜溶液的吸附率分别高达97.1%、93.8%、94.0%和93.3%。BaP和铜在2 h内均没有造成菌体表面形态的明显变化;处理10 mg.L^-1 Cu^2＋及其与BaP的复合污染2 d后,菌体表面会产生突起结构。     

表面活性剂对东北黑土中苯并[a]芘的增溶洗脱作用

研究了4种常见表面活性剂十二烷基苯磺酸钠(SDBS)、十二烷基硫酸钠(SDS)、Tween80和TritonX-100对东北农田黑土土壤中苯并[a]芘的洗脱修复效果。增溶试验结果表明,非离子表面活性剂Tween80和TritonX-100对苯并[a]芘的增溶能力显著高于阴离子表面活性剂SDBS和SDS。4种表面活性剂在单一和混合处理条件下均能不同程度地洗脱土壤中的苯并[a]芘,且洗脱率随着表面活性剂浓度的增加而增大。各表面活性剂洗脱能力大小顺序为Tween80-SDS(3:1)Tween80-SDS(1:1)Tween80-SDS(10:1)Tween80TritonX-100SDSSDBS,当Tween80和SDS质量比为3:1时对两受试土壤中苯并[a]芘的去除率分别达到最高的42.3%和44.8%,可见阴-非离子混合表面活性剂对土壤苯并[a]芘的洗脱效果好于单一表面活性剂。土壤老化70 d后,Tween 80和Tween 80-SDS(3:1)处理对苯并[a]芘的洗脱率相比老化仅14 d的土壤分别下降了20.5%和12.9%,表明土壤老化时间的增加可降低表面活性剂对苯并[a]芘的洗脱去除效果。     

黑麦草对土壤中苯并[a]芘动态变化的影响

本文通过盆栽试验初步研究了黑麦草 (LoliummultiflorumLam)对污染土壤中多环芳烃苯并 [a]芘动态变化的影响。盆栽试验设计 3种苯并 [a]芘处理浓度 ,分别为 1、10、10 0mgkg-1。将苗龄为 1周的黑麦草移植于受苯并 [a]芘污染的土壤中 ,同时设置有相同的苯并 [a]芘处理浓度但不种植物的对照试验。试验在2 0m3 的控温、控光的生长室内进行 ,土壤湿度维持在田间持水量的 6 0 %。通过 12 0d的温室盆栽试验 ,观察到土壤中苯并 [a]芘的可提取浓度随着时间逐渐减少 ,种植黑麦草加快了土壤中可提取态苯并 [a]芘浓度的下降。在 1、10、10 0mgkg-1苯并 [a]芘处理浓度下 ,黑麦草生长的土壤中苯并 [a]芘的减少率分别达 82 3%、74 0 %和 5 5 9%。结果还显示 ,随盆栽时间的延长 ,黑麦草根圈土壤中多酚氧化酶含量提高 ,这可能根圈土壤中可提取态苯并 [a]芘含量降低有关。黑麦草的地上部可以积累苯并 [a]芘 ,变幅在 0 0 6～ 3 6 0mgkg-1。初步认为 ,土壤具有缓解苯并 [a]芘污染的自然本能 ,促进黑麦草生长 ,增强土壤多酚氧化酶活性 ,可提高黑麦草对苯并 [a]芘污染土壤的修复能力。     

土壤中胡敏酸的光化学降解及其对苯并[α]芘结合性质的影响

在不同光照时间、pH和溶解氧浓度下,研究了土壤中胡敏酸经模拟太阳光照射后对苯并[α]芘结合性质的影响.结果表明胡敏酸经模拟太阳光照射后,其与苯并[α]芘的结合系数Koc下降.光化过程中胡敏酸的分子量或SUVA270 与Koc之间有较好的指数回归关系.在pH 4.1～8.0 范围内,在光照后pH条件下测定经光照处理样品的Koc值平均下降34.3% ± 4.1%,在光照前pH条件下测定经光照处理样品的Koc值平均下降33.8% ± 8.3%.溶解氧浓度的升高能加速胡敏酸的光氧化降解过程,与相应的未经光照处理样品相比,在空气饱和与氧气饱和条件下光照后的Koc分别下降37.2% 和47.1%.     

Influence of kaolinite and montmorillonite on benzo[a]pyrene biodegradation by Paracoccus aminovorans HPD-2 and the underlying interface interaction mechanisms

Clay minerals play an important role in biogeochemical cycling.Here,kaolinite and montmorillonite,the two most abundant and widespread clay minerals with typical layered structures,were selected to investigate and compare their effects on the biodegradation of benzo[a]pyrene(BaP)by Paracoccus aminovorans HPD-2 and to investigate the underlying interface mechanisms.Overall,the BaP degradation efficiency was significantly higher 7 d after montmorillonite addition,reaching 68.9%(P<0.05),when compared with that of the control without addition of clay minerals(CK,61.4%);however,the addition of kaolinite significantly reduced the BaP degradation efficiency to 45.8%.This suggests that kaolinite inhibits BaP degradation by inhibiting the growth of strain HPD-2,or its strong hydrophobicity and readily agglomerates in the degradation system,resulting in a decrease in the bio-accessibility of BaP to strain HPD-2.Montmorillonite may buffer some unfavorable factors,and cells may be fixed on the surface of montmorillonite colloidal particles across energy barriers.Furthermore,the adsorption of BaP on montmorillonite may be weakened after swelling,reducing the effect on the bio-accessibility of BaP,thus promoting the biodegradation of BaP by strain HPD-2.The experimental results indicate that differential bacterial growth,BaP bio-accessibility,interface interaction,and the buffering effect may explain the differential effects of the different minerals on polycyclic aromatic hydrocarbon biodegradation.These observations improve our understanding of the mechanisms by which clay minerals,organic pollutants,and degrading bacteria interact during the biodegradation process and provide a theoretical basis for increasing the biodegradation of soil pollutants by native microorganisms under field conditions.     

一株副球菌对污染土壤中多环芳烃的降解研究

从受多环芳烃(PAHs)长期污染的土壤中分离到一株降解PAHs的噬氨副球菌(Paracoccus aminovorans)HPD-2.使用HPD-2的菌液对PAHs污染的土壤进行了2周的生物降解试验,结果表明加入HPD-2能够明显提高土壤中PAHs的降解率.加菌土壤中PAHs的总去除率为22.9%,PAHs各组分的降解率在19.5% ～ 36.2% 之间.其中三环PAHs的降解率最高(36.1%),五环次之(26.0%),四环的最低(20.9%).对土壤微生物的计数结果发现,HPD-2的加入显著提高了土壤中细菌的数量,而对放线菌和真菌的影响不明显.PCR-DGGE分析结果表明,降解过程中HPD-2可能成为土壤中的优势菌.以上结果表明该菌株在PAHs污染土壤的生物降解中具有较好的应用前景.     

Influences of kaolinite and montmorillonite on benzo[a]pyrene biodegradation by Paracoccus aminovorans HPD-2 and the underlying interface interaction mechanisms

Clay minerals play an important role in biogeochemical cycling. Here, kaolinite and montmorillonite, the two most abundant and widespread clay minerals with typical layered structures, were selected to investigate and compare their effects on the biodegradation of benzo[a]pyrene (BaP) by Paracoccus aminovorans HPD-2 and to investigate the underlying interface mechanisms. Overall, the BaP degradation efficiency was significantly higher 7 d after montmorillonite addition, reaching 68.9% (P < 0.05), when compared with that of the control without addition of clay minerals (CK, 61.4%); however, the addition of kaolinite significantly reduced the BaP degradation efficiency to 45.8%. This suggests that kaolinite inhibits BaP degradation by inhibiting the growth of strain HPD-2, or its strong hydrophobicity and readily agglomerates in the degradation system, resulting in a decrease in the bio-accessibility of BaP to strain HPD-2. Montmorillonite may buffer some unfavorable factors, and cells may be fixed on the surface of montmorillonite colloidal particles across energy barriers. Furthermore, the adsorption of BaP on montmorillonite may be weakened after swelling, reducing the effect on the bio-accessibility of BaP, thus promoting the biodegradation of BaP by strain HPD-2. The experimental results indicate that differential bacterial growth, BaP bio-accessibility, interface interaction, and the buffering effect may explain the differential effects of the different minerals on polycyclic aromatic hydrocarbon biodegradation. These observations improve our understanding of the mechanisms by which clay minerals, organic pollutants, and degrading bacteria interact during the biodegradation process and provide a theoretical basis for increasing the biodegradation of soil pollutants by native microorganisms under field conditions.     

长江三角洲地区城市污泥中苯并[a]芘和菲的初步研究

To evaluate the contaminated conditions of benzo[a]pyrene (B[a]P) and phenanthrene (PA) in sludge and to ascertain whether B[a]P limit for land application of sludge exists, the contents of B[a]P and PA in 46 sludge samples from 15 cities in the Yangtze River Delta area of China were determined using high performance liquid chromatography (HPLC) coupled with a fluorescence detector after ultrasonic extraction and silica gel cleanup. B[a]P contents ranged from non-detectable to 1.693 mg kg^-1 dry weight (DW), averaged 0.402 mg kg^-1 DW, and were < 1.0 mg kg^-1 DW in most of the sludge samples. PA was found in all the sludge samples analyzed; its contents ranged between 0.028 and 1.355 mg kg^-1 DW, with an average value of 0.298 mg kg^-1 DW. Most of the sludge samples contained < 0.5 mg kg^-1 DW PA. All 46 municipal sludge samples analyzed in this study showed B[a]P contents < 3.0 mg kg^-1 DW, which is the limit value for sludge applied to agricultural lands in China. The contents and distributions of B[a]P and PA in municipal sludge were related to sludge types, sources and treatment technologies, along with the physical and chemical properties of these pollutants.     

Tolerance,growth and degradation of phenanthrene and benzo[a]pyrene by Rhizobium tropici CIAT 899 in liquid culture medium

Polycyclic aromatic hydrocarbons (PAH) are ubiquitous pollutants that are toxic and recalcitrant to degradation by bacteria. This research evaluated the toxicity of different concentrations [10, 20, 40, 60, 80 and 100 μg mL⁻¹] of phenanthrene (PHE) or benzo[a]pyrene (BaP) on the growth of Rhizobium tropici CIAT899 under in vitro conditions as well as the potential degradation of PHE and BaP by this bacterium. At 24 h, a 40% decrease in Rhizobium growth was observed when exposed to 40 μg mL⁻¹ of either PHE or BaP. Furthermore, bacterial growth was completely inhibited by PHE or BaP applied in 80 and 100 μg mL⁻¹. After 96 h, the growth of R. tropici at 40 μg PHE mL⁻¹ or 60 μg BaP mL⁻¹ was similar to those treatments without PAH. To evaluate R. tropici degrading capabilities, supernatants of cultures with 40 μg PHE mL⁻¹ or 60 μg BaP mL⁻¹ were analyzed by gas chromatography coupled to mass spectrophotometer (GC–MS). R. tropici was able to degrade either PHE or BaP diminishing its concentration in 20% and 25% during the first 24 h, degradation obtained at 120 h was 50% and 45% for PHE or BaP, respectively. This research shows for the first time that R. tropici CIAT 899 grows in liquid culture medium contaminated with PAH, and moreover is able to growth and to degrade either PHE or BaP.     

Selective enrichment of a pyrene degrader population and enhanced pyrene degradation in Bermuda grass rhizosphere

Rhizosphere soil has a more diverse and active microbial community compared to nonvegetated soil. Consequently, the rhizosphere pyrene degrader population (PDP) and pyrene degradation may be enhanced compared to nonvegetated bulk soil (NVB). The objectives of this growth chamber study were to compare (1) Bermuda grass (Cynodon dactylon cv. Guymon) growth in pyrene-contaminated and noncontaminated soils and (2) pyrene degradation and PDP among NVB, Bermuda grass bulk (BB), and Bermuda grass rhizosphere soil (BR). Soils were amended with pyrene at 0 and 500 mg kg^–1, seeded with Bermuda grass, and thinned to two plants per pot 14 days after planting (DAP). Pyrene degradation was evaluated over 63 days. The PDP was enumerated via a most probable number (MPN) procedure at 63 DAP. Bermuda grass root growth was more sensitive to pyrene contamination than shoot growth. Pyrene degradation followed first-order kinetics. Pyrene degradation was significantly greater in BR compared to BB and NVB with rate constants of 0.082, 0.050, and 0.052 day^–1, respectively. The PDPs were 8.01, 7.30, and 6.83 log₁₀ MPN g^–1 dry soil for BR, BB, and NVB, respectively. The largest PDP was in soil with the most rapid pyrene degradation. These results indicate that Bermuda grass can grow in pyrene-contaminated soil and enhance pyrene degradation through a rhizosphere effect.