甲磺隆降解菌FLDA的分离鉴定及其降解特性研究

从生产甲磺隆的农药厂内采取污泥，经驯化富集后筛选到一株能高效降解甲磺隆的细菌FLDA，根据表型特征、生理生化特性及16S rDNA序列同源性分析，将FLDA初步鉴定为假单胞菌（Pseudomonas sp）。该菌能在含甲磺隆（30mgL^-1）的基础盐液体培养基中降解甲磺隆，5d降解率达72．6％，该菌降解甲磺隆的最适pH为7．0，最适温度为30℃，该菌降解甲磺隆的速率和起始接种量呈正相关。酶的定域实验表明，该菌中甲磺隆水解酶为胞内酶。FLDA投加土壤，可提高土壤中甲磺隆的降解速率。     

降解菌S113对甲磺隆污染土壤生物修复作用的研究

在室内条件下,研究了降解菌S113(Methylopila sp.)对甲磺隆污染土壤的修复作用。S113能够以甲磺隆为唯一碳源生长,72h对50mgL-1甲磺隆的降解率达98.38%。投加降解菌S113可显著提高土壤中甲磺隆的降解速率。当甲磺隆浓度为10mgkg-1干土,S113接种量为108个g-1土时,第30天土壤中甲磺隆降解率为76.9%,对照土壤中甲磺隆降解率仅为11.9%。S113降解甲磺隆的速率和接种量呈正相关,当接种量减少为105个g-1干土时,降解菌对甲磺隆的降解作用微弱。在土壤中甲磺隆浓度较低的条件下,S113的降解效果显著,而当土壤中甲磺隆浓度达到50mgkg-1时,甲磺隆降解率仅为39.6%。S113降解土壤中甲磺隆的最适温度为30℃,第30天的降解率可达75.9%。当温度为25℃、20℃时,第30天甲磺隆降解率仅为53.5%和23.9%。S113菌剂灌根,能不同程度地缓解土壤中浓度为40、80μgkg-1的甲磺隆对玉米生长的抑制作用,但当甲磺隆浓度增加到120μgkg-1时,接种S113对药害解除作用不显著。结果表明,人工接种降解菌S113,能有效去除土壤中甲磺隆残留。     

异丙隆在土壤中的淋溶迁移及其影响因素研究

采用室内土壤淋洗柱法,以黄褐土、砂姜黑土和水稻土为供试土壤,研究了异丙隆在土壤中的淋溶迁移行为,探讨了淋溶水量、淋溶水pH值、施药量和添加外源木炭等因素对异丙隆在土壤中淋溶迁移的影响。结果表明,不同土壤中异丙隆淋出率为黄褐土〉砂姜黑土〉水稻土;淋溶水量与异丙隆的淋出率呈正相关,且对淋溶后异丙隆在土层中的分布有明显影响;用不同pH值的淋溶水时,异丙隆的淋出率为pH5〉pH9〉pH7;施加不同药量时,异丙隆的淋出率为10mg〉5mg〉20mg;异丙隆的淋出率随外源木炭添加量的增大而减小,而异丙隆在土壤柱中的滞留量则随着木炭添加量增大而增大,提示添加外源木炭可明显减少异丙隆在土壤中的淋出率,降低异丙隆在土壤中的淋溶深度。     

降解菌HQ-C-01对克百威污染土壤的生物修复

在室内模拟条件下,研究了降解菌HQ-C-01(Pichia anomala)对克百威污染土壤的修复作用及其影响因素,同时研究了克百威及该菌株对土壤微生物的影响。结果表明,克百威降解率与降解菌HQ-C-01接种量呈正相关,降解菌接种量为2.09×108 CFU/g干土时,对土壤中50 mg/kg克百威10天降解率达82.89%;当降解菌接种量低于106 CFU/g干土时,降解菌对克百威的降解效果较弱。土壤含水量显著影响降解菌对克百威的降解率,含水量为600 g/kg时降解效果最好,降解率达85.32%,而当含水量低于200 g/kg时降解效果较差。在温度范围25℃~35℃降解菌对克百威都具有较好的降解效果。不同土壤pH值对降解菌的降解作用有显著影响,在pH值为7时,降解菌对土壤中50 mg/kg克百威10天降解率达85.62%,在较低和较高pH值下,降解效果较差。克百威使用对土壤菌落结构有一定的影响,对土壤真菌具有强烈刺激作用,从而使土壤微生物群落结构发生改变,而降解菌的使用可缓解克百威对土壤微生物的影响,修复受污染土壤。     

Arthrobacter sp. AG1菌株降解土壤中阿特拉津研究

在阿特拉津浓度为50mg/kg干土的黄棕壤、潮土和红壤接种1.5×106CFU/g干土的降解菌Arthrobacter sp. AG1,10天后土壤中的阿特拉津分别降解至1.5、6.6和10mg/kg干土。阿特拉津的降解速率受到土壤性质的影响,但AG1仍能在不满足其生长繁殖要求的pH值的土壤中有效降解酸性土壤中阿特拉津;土壤中水分含量对降解效果影响较大,>20%时降解效果较好;土壤低含水量和低pH值会导致AG1降解阿特拉津的活力下降。不同的接种量对降解效果有一定影响,但105～107CFU/g干土接种量的AG1都能有效发挥降解作用。AG1降解完土壤中的阿特拉津后,在土壤含水量分别为5%和15%的情况下能长期保持降解活性,对60天后第2次施入黄棕壤和潮土中的50mg/kg阿特拉津4天时降解效率在65%以上。     

复合菌群的构建及其对石油污染土壤修复的研究

从石油污染土壤中富集分离、筛选出3株高效降解石油的微生物菌株,通过生理生化特性研究及16SrRNA基因序列分析,确定3株菌均属于红球菌属（Rhodococcus sp）,研究和比较了它们与实验室保存的4株菌（分别属于Gordonia sp,Comamonas sp,Pesudomonas sp）降解石油的能力。这7株菌株对石油的不同组分具有不同的降解能力,对7株菌进行不同的组合用以研究复合菌群对石油的降解。结果表明,由两株Rhodococcus sp,一株Gordonia sp和一株Pesudomonas sp组成的复合菌群D,降解石油的能力超过任何单一菌株和其他组合菌群。混合菌群D在5d的培养中能降解70.3%的石油总量和71.4%的芳香化合物。混合菌群D能降解99.8%的C13-19烷烃,92.6%的C20-26烷烃,82.2%的C27-32烷烃以及90.2%的植烷。在实验室模拟条件下,对土壤中石油的降解率达到50%以上。降解土壤中石油的最适温度为10～30℃、pH值为6.5～9.5,接种量需要在106CFU·g-1以上。     

一株毒死蜱降解菌株Sphingomonas sp.Dsp-2的分离鉴定及降解特性

从长期受毒死蜱污染的污水处理池中分离到一株毒死蜱高效降解菌株,命名为Dsp-2。经生理生化和16S rDNA序列同源性分析,鉴定其为鞘胺醇单胞菌属(Sphingomonas sp.)细菌。该菌株能在24 h内完全降解100 mg L-1的毒死蜱,降解特性的研究表明:随着农药浓度的加大,绝对降解量也增大,但高浓度的毒死蜱会导致不能完全降解;起始接种量和降解毒死蜱的速率呈正相关;外加氮源营养能够明显促进降解;1 mmol L-1的Fe3 和Ni2 等对其降解性能有抑制作用。研究了Dsp-2在土壤中降解毒死蜱的效果。结果表明,Dsp-2在三种供试土壤中都能有效的降解毒死蜱,其中在潮土中降解的速率最快,且当毒死蜱的浓度范围在1~100 mg kg-1内Dsp-2都能有效的降解毒死蜱,7 d降解率达到85%~98%。     

纳米Fe^0与微生物协同降解土壤中PCB77

以红壤、黄褐土和砂姜黑土为供试土壤,以3,3′,4,4′-四氯联苯（PCB77）为目标化合物,进行了纳米Fe0、微生物以及联合体系降解土壤中PCB77的动力学研究。结果表明,土壤中PCB77自然降解率低,投加纳米Fe0和微生物均能显著加快土壤中PCB77降解速率,降解过程可用一级反应动力学方程拟合。当PCB77初始浓度为4mg·kg-1,纳米Fe0投加量为20mg·g-1,PCB77在红壤、黄褐土和砂姜黑土中反应速率常数k分别为0.0205d-1、0.0165d-1和0.0145d-1;通过富集培养的方法从污染土壤中分离出一株多氯联苯降解菌,初步鉴定该菌株为Pseudomonas sp.,当降解菌投加量为2×108cfu·g-1时,PCB77在3种土壤中反应速率常数分别为0.0136d-1、0.0094d-1和0.0124d-1;当同时投加20mg·g-1纳米Fe0和2×108cfu·g-1降解菌时,其反应速率常数分别为0.0264d-1、0.0218d-1和0.0232d-1。纳米Fe0与微生物协同降解的效果要明显优于纳米Fe0和微生物的单一体系。     

木霉T12菌株对多菌灵的降解特性及生物修复

从耐药性木霉菌株的诱变选育过程中,得到一株能在含多菌灵2000mgL-1培养基上生长的变异菌株T12。该菌株在以多菌灵为唯一碳源的无机盐培养基中,于25℃、200rmin-1振荡培养5d,对多菌灵的降解率达到73.1%。在pH6.0、温度25℃、5%接种量和加入0.5%酵母粉为最适降解条件下,该菌株对多菌灵的降解率达到92.1%。对原土壤、自然风干土壤和高温烘干土壤中的多菌灵进行室内降解实验,在25℃～28℃,5%接种量,15%含水量的条件下处理10d,对多菌灵的降解率分别达到79.7%、76.5%和70.5%。研究结果为该菌株在土壤生物修复中的应用提供了科学依据。     

4株苯磺隆降解菌的分离鉴定及其生长特性

从长期受农药苯磺隆污染的土壤中通过采用富集培养分离技术得到4株以苯磺隆为唯一碳源生长的细菌,分别将其命名为B1、B2、B3和B4。通过观察这4种菌株的形态学特征,研究其生理生化特性以及分析其16S rDNA序列,初步鉴定菌株B1为铜绿假单胞菌(Pseudomonas aeruginosa),B2为戴尔福特菌(Delftia sp.),B3为微杆菌(Microbacterium sp.),B4为产碱杆菌(Alcaligenes sp.)。并通过研究温度、初始pH值、接种量、苯磺隆初始浓度、培养基体积、氮源、碳源、Mg2+浓度等因素对4种菌株生长情况的影响,确定了菌株的最佳生长条件。结果显示,B1菌株的最适温度为35℃,其他3株菌株均为30℃。菌株B3最适pH为8.0,其余3株菌株均为pH7.0。B1和B3菌株最适接种量为15%,B2和B4最适接种量为10%。菌株B3最适苯磺隆初始浓度为100mg·L-1,其余菌株最适苯磺隆初始浓度均为200mg·L-1。4株菌株最适培养基体积均为75mL,最适氮源均为硝酸铵,最适碳源均为葡萄糖。B2菌株最适Mg2+浓度为100mg·L-1,其余3株菌株均为200mg·L-1。B1和B4菌株最适NaCl浓度为20g·L-1,B2菌株NaCl浓度为5~30g·L-1,B3菌株最适NaCl浓度为50g·L-1。该结果为利用微生物对农药苯磺隆污染的土壤进行原位生物修复提供理论依据。     

Spatial heterogeneity in the metabolism and dynamics of isoproturon degrading microbial communities in soil

The characteristics of isoproturon [3-(4-isopropylphenyl)-1,1-dimethylurea] metabolism were investigated in soil taken from two transects within a single field. Along transect 1, complete degradation of the parent compound occurred within 18 days, and over 40% of ring C had been metabolised after 65 days. In these soils, both side chain and ring metabolism had a short lag phase, followed by a period of rapid degradation. Along transect 2, the rate of side chain metabolism was highly variable, and 20% of ring C had been metabolised after 65 days. The dynamics of isoproturon ring C metabolism were typical of cometabolic degradation, even at sites in which enhanced side chain metabolism occurred. Isoproturon degrading organisms were found in similar numbers in soil from the two transects prior to isoproturon application. In soils from transect 1, there was considerable proliferation of degrader organisms during the lag phase, in which 40% of the isoproturon was degraded. In most soils from transect 2, there had been no proliferation of isoproturon metabolising organisms at the point of 40% metabolism. Before enhanced degradation could develop, there was clearly a requirement for the isoproturon metabolising community to reach a threshold size. Immobilisation of isoproturon ring C into the microbial biomass and formation of bound residues was lower in soil from transect 2 relative to soil from transect 1. We conclude that the in-field spatial heterogeneity of isoproturon side chain and ring metabolism, the formation of bound residues and the immobilisation of pesticide residues in the biomass, results from variation in the development and significance of growth linked and cometabolic degradation.     

土壤和土壤悬液中1,2,4-三氯苯的微生物强化降解

Inoculating soil with an adapted microbial community is a more effective bioaugmentation approach than inoculation with pure strains in bioremediation.However,information on the potential of different inocula from sites with varying contamination levels and pollution histories in soil remediation is lacking.The objective of the study was to investigate the potential of adapted microorganisms in soil inocula,with different contamination levels and pollution histories,to degrade 1,2,4-trichlorobenzene (1,2,4-TCB).Three different soils from chlorobenzene-contaminated sites were inoculated into agricultural soils and soil suspension cultures spiked with 1,2,4-TCB.The results showed that 36.52% of the initially applied 1,2,4-TCB was present in the non-inoculated soil,whereas about 19.00% of 1,2,4-TCB was present in the agricultural soils inoculated with contaminated soils after 28 days of incubation.The soils inoculated with adapted microbial biomass (in the soil inocula) showed higher respiration and lower 1,2,4-TCB volatilization than the non-inoculated soils,suggesting the existence of 1,2,4-TCB adapted degraders in the contaminated soils used for inoculation.It was further confirmed in the contaminated soil suspension cultures that the concentration of inorganic chloride ions increased continuously over the entire experimental period.Higher contamination of the inocula led not only to higher degradation potential but also to higher residue formation.However,even inocula of low-level contamination were effective in enhancing the degradation of 1,2,4-TCB.Therefore,applying adapted microorganisms in the form of soil inocula,especially with lower contamination levels,could be an effective and environment-friendly strategy for soil remediation.     

Harsh summer conditions caused structural and specific functional changes of microbial communities in an arable soil

The mineralization of the herbicide 3-(4-isopropylphenyl)-1,1-dimethylurea (isoproturon) was reduced after the dry and hot summer 2003 in a soil profile placed in a field lysimeter. A different isoproturon mineralization pattern remained in soil material taken at two different soil depths (0–5 cm and 15–20 cm), although soil material was re-equilibrated at adequate climatic conditions. Special soil microcosms were designed to determine if the changes in this special soil function 'isoproturon mineralization' were related to the climatic scenario of summer 2003. These microcosms were filled with lysimeter soil from the 15–20 cm depth and the temperature and dryness of summer 2003 were simulated. Afterwards, soil samples were taken from the microcosms and re-equilibrated under controlled conditions for 4 weeks. Subsequently, isoproturon mineralization was investigated. The soil microbial community reduced drastically its original capability of isoproturon mineralization in the course of the model experiments.
Analysis of 16S-rRNA by denaturing gel gradient electrophoresis (DGGE) revealed substantial differences in the band patterns of the bacterial communities from both depths of the field lysimeter soil and from the soil incubated in microcosms. The different soil microbial biomass determined by microcalorimetry reinforced these results. In conclusion, the factors higher temperature and smaller soil moisture content generated important and enduring changes in the microbial community structure and therefore in specific soil functions of the community, as shown here by the function of isoproturon degradation. Results are discussed in connection with environmental conditions and conservation tillage.     

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。     

阿维菌素在土壤中的降解和高效降解菌的筛选

运用恒温培养法研究了阿维菌素在不同土壤中的降解动力学。结果表明 ,土壤有机质、土壤温度和农药浓度对阿维菌素的降解有较大影响 ,这可能和土壤微生物有关。从试验土壤中分离到一株高效降解阿维菌素的菌株 ,经 16SrDNA鉴定为嗜麦芽寡养单胞菌 (Stenotrophomonasmaltrophilia)。土壤接种该优势菌后有助于加快阿维菌素的降解     

Bioremedial potential of fenamiphos and chlorpyrifos degrading isolates: Influence of different environmental conditions

Previously isolated bacterial strains for chlorpyrifos and fenamiphos degradation were used to examine their potential as bioremedial agents in soils and water containing pesticide residues. Both, chlorpyrifos-degrading Enterobacter sp and fenamiphos-degrading consortium rapidly degraded pesticides when inoculated into natural and sterile water and soils. Degradation rate was slower in lower pH soils in comparison with natural and alkaline soils. Soil organic matter had no impact on pesticide degrading ability of isolates. Soil moisture <40% of maximum water-holding capacity slowed down degradation rate. The bacterial isolates were able to rapidly degrade fenamiphos and chlorpyrifos between 15 and 35 °C but their degradation ability was sharply reduced at 5 and 50 °C. Both groups of bacterial systems were also able to remove a range of pesticide degradation. An inoculum density of 10⁴ cells g⁻¹ of soil was required for initiating rapid growth and degradation. Ageing of pesticide in soils prior to inoculation produced contrasting results. Ageing of fenamiphos had no impact on subsequent degradation by the inoculated consortium. However, degradation of chlorpyrifos by Enterobacter sp after aging resulted in persistence of ∼10% of pesticide in soil matrix. Higher K_oc value of chlorpyrifos may have resulted in a lack of bioavailability of a smaller percentage of chlorpyrifos to degrading bacteria. Overall, this paper confirms bioremedial potential of a fenamiphos degrading consortium and a chlorpyrifos degrading bacterium under different soil and water characteristics.     

Biodegradation of buprofezin by Rhodococcus sp. strain YL-1 isolated from rice field soil

A buprofezin-degrading bacterium, YL-1, was isolated from rice field soil. YL-1 was identified as Rhodococcus sp. on the basis of the comparative analysis of 16S rDNA sequences. The strain could use buprofezin as the sole source of carbon and nitrogen for growth and was able to degrade 92.4% of 50 mg L(-1) buprofezin within 48 h in liquid culture. During the degradation of buprofezin, four possible metabolites, 2-tert-butylimino-3-isopropyl-1,3,5-thiadiazinan-4-one, N-tert-butyl-thioformimidic acid formylaminomethyl ester, 2-isothiocyanato-2-methyl-propane, and 2-isothiocyanato-propane, were identified using gas chromatography-mass spectrometry (GC-MS) analysis. The catechol 2,3-dioxygenase activity was strongly induced during the degradation of buprofezin. A novel microbial biodegradation pathway for buprofezin was proposed on the basis of these metabolites. The inoculation of soils treated with buprofezin with strain YL-1 resulted in a higher degradation rate than that observed in noninoculated soils, indicating that strain YL-1 has the potential to be used in the bioremediation of buprofezin-contaminated environments.     

Microbial degradation of fomesafen by a newly isolated strain Pseudomonas zeshuii BY-1 and the biochemical degradation pathway

Fomesafen is a diphenyl ether herbicide used to control the growth of broadleaf weeds in bean fields. Although the degradation of fomesafen in soils was thought to occur primarily by microbial activity, little was known about the kinetic and metabolic behaviors of this herbicide. This paper reported the capability of the newly isolated strain Pseudomonas zeshuii BY-1 to use fomesafen as the sole source of carbon in pure culture for its growth. Up to 88.7% of 50 mg of L(-1) fomesafen was degraded by this bacterium in mineral medium within 3 days. Strain BY-1 could also degrade other diphenyl ethers, including lactofen, acifluorfen, and fluoroglycofen. During the fomesafen degradation, five metabolites were detected and identified by liquid chromatography-mass spectrometry and tandem mass spectrometry. The primary degradation pathway of fomesafen might be the reduction of the nitro group to an amino group, followed by the acetylation of the amino derivative, dechlorination, and cleavage of the S-N bond. The addition of the BY-1 stain into soils treated with fomesafen resulted in a higher degradation rate than that observed in uninoculated soils, and the bacteria community in contaminated soil recovered after inoculation of the BY-1 stain. On the basis of these results, strain P. zeshuii BY-1 has the potential to be used in the bioremediation of fomesafen-contaminated soils.