Biodegradation of anthracene and phenanthrene by bacteria isolated from oil-contaminated soil of Bangladesh

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous environmental pollutants entering into the environment through natural and anthropogenic activities. Owing to their toxicity to various life forms including humans, detoxification of PAHs is crucial to reduce their effects on cells. In this study, we have isolated two bacteria capable of degrading two common PAHs, anthracene and phenanthrene, from contaminated soil samples by using selective enrichment culture supplemented with test PAHs as the sole source of carbon and energy. The isolated bacteria were identified and affiliated as Pseudomonas aeruginosa strain KD and Stenotrophomonas maltophilia strain RC based on their 16s rRNA gene sequences. The degradation of anthracene and phenanthrene was estimated indirectly by measuring the decolourisation extent of a redox indicator, 2, 6-dichlorophenolindophenol, incorporated into PAH-supplemented mineral salt media. In the case of anthracene, ≥90% decolourisation was recorded at 20 and 48 days for P. aeruginosa and S. Maltophilia, respectively. On the other hand, ≥94% decolourisation was recorded at 56 and 52 days for P. aeruginosa and S. maltophilia, respectively during the utilisation of phenanthrene.     

Biodegradation of paclobutrazol by a microbial consortium isolated from industrially contaminated sediment

Biodegradability of the plant growth retardant paclobutrazol by a microbial consortium in which Pseudomonas was the predominant strain was investigated in batch culture. The consortium which had been isolated from an industrially contaminated sediment was proven to be useful for the treatment of effluents containing paclobutrazol. Paclobutrazol was degraded by the pure isolated strain of Pseudomonas sp. as well as the microbial consortium. Paclobutrazol was utilized as the sole source of carbon and energy. Sixty percent of the paclobutrazol was degraded by the microbial consortium from an initial concentration of 54 mg L^?1 within 48 h and more than 98% of an initial concentration of 3.4 mg L^?1 was degraded within 36 h. The optimum temperature and pH were determined to be 30°C and 7.0, respectively. A pure strain of a bacterium, isolated from the enrichment culture was identified as Pseudomonas sp. The microbial consortium was tolerant of high pH and could degrade paclobutrazol faster than the pure strain. The degradation rate of this plant growth regulator in an aerobic environment was greater than that under anaerobic conditions.     

生物表面活性剂鼠李糖脂对PCBs污染土壤的修复作用研究

利用生物表面活性鼠李糖脂（RL）洗脱土壤,再通过紫外光预照射与生物降解协同去除洗脱液中的PCBs的组合方法对多氯联苯（PCBs）污染土壤进行修复,旨在研究RL在修复PCBs污染土壤中的作用及其机理。结果表明,RL对PCBs的洗脱具有显著的促进作用,PCBs的总洗脱率与RL的质量浓度呈正相关。当洗脱液中加入2 000 mg.L-1的RL,在3次批洗脱后,人工污染土样和陈化土样的PCBs总洗脱率分别达到了90.1%和47.1%。PCBs降解菌P.LB400在以RL或联苯为碳源的驯化培养基中均能够快速生长。当土壤洗脱液中的PCBs被P.LB400的生长细胞菌液降解时,RL对PCBs的生物降解具有显著的促进作用;而在P.LB400的休眠细胞降解体系中,RL对PCBs的生物降解有一定的抑制作用。紫外光预照射对土壤洗脱液中PCBs的生物降解有一定的促进作用。紫外光预照射和生物降解的耦合有利于提高PCBs的降解速率。     

Characterisation and identification of carbofuran-utilising bacteria isolated from agricultural soil

A total of 96 bacterial cultures were isolated from soil. Seventeen bacterial isolates were selected following their cultivation on solid media containing 100 mg · L^?1 carbofuran as the sole source of carbon and nitrogen. Of the 17 isolates, 10F, 11M, 17N, 23B and 26M were specifically chosen because of their relatively higher growth efficiency and genetic diversity based on Box-polymerase chain reaction analysis. These bacterial cultures had 16S rRNA gene sequences that were most similar to Acinetobacter baumannii (10F), Agrobacterium tumefaciens (11M), Ochrobactrum anthropi (17N), Escherichia coli (23B) and Agrobacterium tumefaciens (26M) with 97, 95, 93, 95 and 94% similarity in their 16S rDNA gene sequence, respectively. Degradation rates of carbofuran in soil inoculated with these isolates were 1.9, 1.5, 1.6, 1.7 and 1.6 times, respectively, faster in comparison with uninoculated soil after 10 days of incubation. The maximum degradation rates of carbofuran (45 and 91%) were detected in soil inoculated with A. baumannii (10F) after 10 and 20 days’ incubation, respectively. These data indicate that these isolates may have the potential for use in bioremediation of pesticide contaminated soil.     

一株产生物絮凝剂的Bacillus sp.的分离与鉴定

从重庆城南污水处理厂的活性污泥中分离筛选到一株絮凝剂产生菌,革兰氏染色呈阳性,好氧,有芽孢,可运动,短杆;分离菌的基因组DNA(G +C)mol%含量测定为5 0 .6 % ,16SrRNA序列分析表明,分离筛选出的絮凝剂产生菌的16SrRNA基因序列与Bacillus菌有高度同源性,从而确定絮凝剂产生菌RL - 2归于Bacillusbrevis菌属。絮凝实验表明:在有CaCl2 存在时用Bacillussp .RL - 2生物絮凝剂处理土壤悬液时用量少(10 0mL样品只需2mL絮凝剂) ,絮体形成快,矾花大,泥少;体系温度对活性影响很小,且在酸性与碱性条件下,絮凝活性均很高     

Bioaccumulation of organochlorine pesticides from contaminated soil by cattle

The study aimed to quantitate the uptake by beef cattle of organochlorine pesticides in from soil. Although such uptake is known to occur, uptake rates have not been accurately measured for these animals and cannot be inferred from data obtained with dairy cattle. In this study, cattle received a diet, of a standard feedlot ration supplemented with organochlorine-pesticide (aldrin?+?dieldrin) contaminated soil. The levels of pesticides in the feed would approximate uptake through normal grazing. Samples of the subcutaneous fat were removed periodically and analysed for pesticide content. Uptake of dieldrin (but not other pesticides present in the soil) was rapid and substantial. Uptake from sandy soils was more efficient than that from clay soils. The experiment was terminated after 8 weeks, because of the high levels of dieldrin in the tissues. Depuration of the accumulated pesticides was a slow and complex process, requiring a longer period than accumulation.     

土壤和地下水中多环芳烃生物降解研究进展

多环芳烃是一类普遍存在于环境中的难降解的危险性"三致"有机污染物。受污染的土壤和地下水中的多环芳烃,生物降解是其归宿的主要途径。研究表明,对于土壤中低分子量多环芳烃类化合物,微生物一般以唯一碳源方式代谢;而大多数细菌和真菌对四环或四环以上的多环芳烃的降解作用一般以共代谢方式开始。文章重点论述了多环芳烃的来源、降解多环芳烃的微生物、生物降解机理、影响生物降解的因素以及生物修复方法。认为今后的研究方向是高分子量多环芳烃的降解机理与降解途径,基因工程技术在多环芳烃生物降解方面的应用,以及生物表面活性剂产生的机理及其在实际处理中的应用等。     

蜡状芽孢杆菌不同菌株降解有机磷农药的特性分析

筛选分离降解微生物解决有机磷农药残留给水体和土壤环境带来的污染问题是一项可行的生物修复技术。采用富集培养和定时取样分析有机磷农药残留的方法,分离驯化出三株能够降解有机磷农药的细菌,研究了其形态特征和生理生化特性并对其16SrDNA序列进行了分析,同时比较了三菌株对甲基对硫磷（Methyl－parathion）、毒死蜱（Chlorpyrifos）和_二唑磷（Triazophos）的降解特性。结果表明：通过富集培养得到10菌株具有降解甲基对硫磷和毒死蜱的能力,比较确定HY-1、HY-2和HY-4三菌株作为研究对象,经鉴定为蜡状芽孢杆菌（Bacilluscereus）的不同菌株,三菌株在Genbank上的登录号分别为：eu915687、eu915686和eu915688。在甲基对硫磷质量浓度为50mg·L-1时,三菌株72h的降解率分别为91．7％、87．7％与92．4％．降解率无显著性差异（P〉0．05）,当甲基对硫磷质量浓度增加到100mg·L-1时,三菌株对甲基对硫磷的降解率有所下降,其中HY-2对甲基对硫磷的降解率下降最大达23％,且和其他两菌株有显著性差异（P〈0．05o三菌株72h对100mg·L-1毒死蜱的降解率分别达到64．8％、53．7％和56．5％,在不同的毒死蜱初始质量浓度下,HY-1和HY-4两菌株对毒死蜱的降解率无显著性差异（P〉0．05）,HY-2与HY-1、HY-4两菌株有显著性差异（P〈0．05o三菌株对三唑磷的降解率均较低,其中HY-2对初始质量浓度为100mg·L-1三唑磷的降解率最高仅为20．7％,其余两菌株对三唑磷的降解率比HY-2低且无显著性差异（P〉0．05o可以得出本研究分离得到的蜡状芽孢杆菌不同菌株对有机磷农药的降解存在多态性。     

Removal of high concentrations of polycyclic aromatic hydrocarbons from contaminated soil by biodiesel

Solubilizing experiments were carried out to evaluate the ability of biodiesel to remove polycyclic aromatic hydrocarbons (PAHs) from highly contaminated manufactured gas plant (MGP) and PAHs spiked soils with hydroxypropyl-β-cyclodextrin (HPCD) and tween 80 as comparisons. Biodiesel displayed the highest solubilities of phenanthrene (420.7 mg·L^-1), pyrene (541.0 mg·L^-1), and benzo(a)pyrene (436.3 mg·L^-1). These corresponded to several fold increases relative to 10% HPCD and tween 80. Biodiesel showed a good efficiency for PAH removal from the spiked and MGP soils for both low molecular weight and high molecular weight PAHs at high concentrations. Biodiesel was the best agent for PAH removal from the spiked soils as compared with HPCD and tween 80; as over 77.9% of individual PAH were removed by biodiesel. Tween 80 also showed comparable capability with biodiesel for PAH solubilization at a concentration of 10% for the spiked soils. Biodiesel solubilized a wider range of PAHs as compared to HPCD and tween 80 for the MPG soils. At PAH concentrations of 229.6 and 996.9 mg·kg^-1, biodiesel showed obvious advantage over the 10% HPCD and tween 80, because it removed higher than 80% of total PAH. In this study, a significant difference between PAH removals from the spiked and field MGP soils was observed; PAH removals from the MGP soil by HPCD and tween 80 were much lower than those from the spiked soil. These results demonstrate that the potential for utilizing biodiesel for remediation of highly PAH-contaminated soil has been established.     

废铁屑修复汞污染土壤的实验研究

应用工厂废弃物—铁屑做原料,探讨不同铁屑投加量对修复不同土壤类型、不同程度汞污染土壤的能力,并与修复水体汞污染作了对比;另外,首次将淋滤法与国际上新兴的土壤、地下水修复技术-PRB技术结合,研究其修复土壤汞污染的效果。结果表明,废铁屑可很快去除水体中的汞,去除率可达93%;在铁屑存在下,能一定程度预防和修复土壤汞污染,在相同水体汞污染程度条件下,时间是影响土壤汞污染程度的主要因素;在CaCl2淋洗液作用下的模拟PRB实验中,土壤有效汞含量降低,同时淋洗液中汞质量浓度低于1μg·L-1。因此,用废铁屑能有效预防和修复土壤汞污染,且将淋滤法与PRB技术联用,与将铁屑直接撒入土壤中相比效果好得多,且不会引起后续的水体二次污染,是一种很有应用潜力、值得深入研究的土壤修复方法。     

Chelator-induced bioextraction of heavy metals from artificially contaminated soil by mushroom (Coprinus comatus)

This study investigated the potential use of chelator-induced bioextraction of heavy metals from soil by Coprinus comatus in a pot experiment. Two production waves of the mushroom were obtained to determine biomass and metal concentration. The application of ethylenediaminetetraacetic acid (EDTA) or nitrilotriacetate (NTA) slightly inhibited the growth of C. comatus, but significantly enhanced the accumulation of heavy metals in fruiting bodies compared with the control. The highest concentrations of Pb, Cu and Cd reached 900.60, 783.61 and 23.64 mg·kg^?1 in a single pot, respectively. However, application of citric acid (CA) had no effect on metal uptake. Moreover, chelators applied to soil after fructification increased the dry biomass and metal concentration in fruiting bodies compared with those applied before sowing mycelia. EDTA was more efficient for inducing metal uptake by C. comatus than NTA or CA, and split applications of EDTA after fructification resulted in the highest total metal uptake by mushroom, i.e. 19.08±2.84, 17.57±0.69 and 0.55±0.06 mg for Pb, Cu and Cd, respectively, which were 130, 12 and 5 times values obtained with the control. Interestingly, many mushrooms turned blue after soil had been treated with chelator, indicating that these mushrooms are rich in Cu.     

十溴联苯醚(BDE209)对蜡状芽胞杆菌吸附/释放金属离子的影响

针对重金属及多溴联苯醚(PBDEs)复合污染问题,以蜡状芽孢杆菌(Bacillus cereus)复合菌为吸附材料,研究了十溴联苯醚(BDE209)对蜡状芽孢杆菌复合菌吸附及释放金属离子的影响.结果表明,蜡状芽孢杆菌复合菌对低浓度的Pb(II)(0.712mg·L-1)、Zn(II)(4.844mg·L-1)具有快速、高效、稳定的吸附能力,最高吸附率分别达到98.31%和97.83%;对低浓度的Cu(II)(1.915mg·L-1)也能起到一定的去除作用,吸附率最高可达59.90%.复合菌吸附重金属离子的同时,释放了其他金属离子:Ca(II)、K(I)、Na(I),且释放总量大于吸附总量.不同浓度的BDE209对复合菌吸附重金属产生不同的影响.1mg·L-1的BDE209基本表现为促进复合菌对重金属的吸附;而10mg·L-1的BDE209在0～4h之间促进复合菌对Pb(II)、Zn(II)的吸附,在反应4h后明显抑制吸附作用,而对Cu(II)则始终表现为促进作用.BDE209对金属离子释放的影响主要体现在反应的初始阶段(0～4h),表现为高浓度的BDE209减缓K(I)、Na(I)的释放.     

Biodegradation of 2-methylisoborneol by bacteria enriched from biological activated carbon

One of the most common taste and odour compounds (TOCs) in drinking water is 2-methylisoborneol (2-MIB) which cannot be readily removed by conventional water treatments. Four bacterial strains for degrading 2-MIB were isolated from the surface of a biological activated carbon filter, and were characterized as Micrococcus spp., Flavobacterium spp., Brevibacterium spp. and Pseudomonas spp. based on 16S rRNA analysis. The removal efficiencies of 2-MIB with initial concentrations of 515 ng·L^-1 were 98.4%, 96.3%, 95.0%, and 92.8% for Micrococcus spp., Flavobacterium spp., Brevibacterium spp. and Pseudomonas spp., respectively. These removal efficiencies were slightly higher than those with initial concentration at 4.2 mg·L^-1 (86.1%, 84.4%, 86.7% and 86.0%, respectively). The kinetic model showed that biodegradation of 2-MIB at an initial dose of 4.2 mg·L^-1 was a pseudo-first-order reaction, with rate constants of 0.287, 0.277, 0.281, and 0.294 d^-1, respectively. These degraders decomposed 2-MIB to form 2-methylenebornane and 2-methyl-2-bornane as the products.     

Combined use of earthworm (Alma millsoni) and bacterium (Bacillus sp.) improved the bioremediation of spent engine oil contaminated soil

This study was carried out to assess the effectiveness of the combined use of earthworm (Alma millsoni) and bacterium (Bacillus sp.) in the bioremediation of spent engine oil (SEO) contaminated soils. A. millisoni were collected from the Botanical Garden of University of Ibadan, Nigeria. The stock culture of hydrocarbonoclastic Bacillus sp. was used for the bioremediation study. A set-up of eight pots containing 1000?g soil sample and 20?g of cow dung were mixed with 100, 75, 50 and 0?mL SEO respectively. Each of the set up was subjected to bioremediation agents; A. millisoni alone, Bacillus sp alone, A. millisoni and Bacillus sp, no treatment (control) in duplicate. Treatment of 100?ml SEO contaminated soil with combined A. millisoni and Bacillus sp resulted in significantly (P? bacterium > vermi-remediating agents. Earthworms exposed to 100?mLSEO-contaminated soil had higher CAT, SOD, and GPx activities compared to the control. Findings indicated that A. millisoni with Bacillus sp. can synergistically improve bioremediation of SEO contaminated soils.     

黄棕壤镉污染的电动修复及肥力的提升

近些年来,有关施加电场以去除土壤中污染物的报道较多,但是关于电动力学对土壤肥力的影响方面的报道很少。文章研究了电动力学(1V·cm-1)对土壤肥力的影响。修复前后测量的指标有土壤有机质(SOM)、pH、电导率(EC)、氧化还原电位(ORP)、有效氮、速效磷、速效钾。镉质量分数约为100mg·kg-1的模拟污染土壤经电动力学处理60h后,镉的平均去除效率为71%。土壤中的有效氮、速效磷和速效钾分别平均增加38.49%、47.55%和26.81%,同时TOC也增加了7.96%。总之,电动力学修复后,土壤的肥力有了明显的提升。研究结果表明,电动力学在有效去除土壤中镉的同时还能提升土壤的肥力,是一种可行的土壤修复技术。     

Capacity of simultaneous removal of zinc and cadmium from contaminated media, by two microalgae isolated from a polluted site

Several aquatic environments have been contaminated with heavy metals dumped via industrial effluents. Numerous studies have been published regarding the removal of single metals from aqueous solutions by microalgal biomass. However, such studies do not reflect the actual problem associated with industrial effluents because usually more than one metal species is present. Here we studied the biosorption capacity of Zn²⁺ and Cd²⁺ as single- and binary-metal systems by two microalgae, Scenedesmus obliquus and Desmodesmus pleiomorphus, isolated from a polluted site in Northern Portugal. For each metal independently, D. pleiomorphus showed a higher metal sorption capacity than S. obliquus, at concentrations ranging from 60 to 300 mg/l (except 150 mg_Cd/l). Maximum amounts of Zn²⁺ and Cd²⁺ removed were 22.3 and 60.8 mg/g by S. obliquus, and 83.1 and 58.6 mg/g by D. pleiomorphus. In binary-metal solutions, S. obliquus was in general able to remove Zn²⁺ to higher extents than Cd²⁺, whereas the opposite was observed with D. pleiomorphus. The simultaneous uptake of Zn²⁺ and Cd²⁺ by both microalgae was considerably lower than that of their single-metal counterparts, at equivalent concentrations. Although microalgal uptake from binary-metal solutions was lower than from single-metal ones, the wild microalgae selected were able to efficiently take up mixtures of Zn²⁺ and Cd²⁺ up to 300 mg/l of both metals—thus materializing a promising bioremediation vector for polluted waters.     

Biodegradation of crude petroleum and petroleum products by fungi isolated from two oil seeds (melon and soybean)

Crude petroleum oil degrading fungi were isolated from two oil seeds, Cucumeropsis mannii (melon) and Glycine max (soybean) seeds in the presence and absence of petroleum fumes. An assessment of the relative ability of each fungus to degrade crude petroleum, diesel and kerosene on minimum salt solution was done using change in optical density read on spectrophotometer. Twenty-one fungal species (14 genera) were isolated altogether during this experiment. These include eight species of Aspergillus; one species each of Botryodiplodia, Bipolaris, Cladosporium, Cunnighamella, Dreschlera, Fusarium, Helminthosporium, Macrophomina, Mucor, Paeciliomyces, Penicillium, Rhizopus and Talariomyces. It was evident that most of the fungi tested were able to biodegrade the crude petroleum oil, though at different rates. Bipolaris had a low rate of biodegradation of the petroleum oil of all the fungal species isolated Botryodiplodia theobromae had the highest degrading ability on the crude oil, while Aspergillus flavus had the least after 40 days of incubation. Aspergillus flavus had the highest ability to biodegrade diesel while A. niger had the least ability. In kerosene, Macrophomina phaseolina had the highest ability while A. niger had the least ability to biodegrade it. There was fluctuation in the growth pattern of the fungi in the petroleum oil medium. The implication of these are discussed.     

Reduction of hexavalent chromium using fungi and bacteria isolated from contaminated soil and water samples

The reductive adsorption of hexavalent chromium (Cr⁶⁺) using six indigenous microorganisms isolated from contaminated soil and water samples was investigated. Quantification of Cr⁶⁺ reduction was determined using the 1,5-diphenylcarbazide method followed measuring the absorbance at OD₅₄₀. Bacterial isolates identified as Klebsiella pneumoniae, Bacillus firmus and Mycobacterium sp. were capable of absorbing Cr⁶⁺ efficiently into their biomass, whereas the fungal isolates, Aspergillus flavus, Aspergillus sp. and A. niger were capable of transforming Cr⁶⁺ to Cr³⁺ relative to cell-wall-binding properties. Infrared spectral analysis of functional groups showed that ?OH, ?NH₂ and C?O with conjugated ?NH were the binding groups responsible for adsorption of Cr⁶⁺ within the biomass of isolates. The data highlight the promising biotechnological application of these isolates in removing carcinogenic and mutagenic Cr⁶⁺ from contaminated ecosystems.     

Remediation rates and translocation of heavy metals from contaminated soil through Parthenium hysterophorus

The potential translocation of heavy metals by Parthenium hysterophorus over 30 and 90 days and its effect on biomass, chlorophyll content and photosynthetic activity were studied on 0, 10, 25, 50 and 100% fly-ash-amended soil (FAS). The results showed a decrease in chlorophyll content and photosynthetic area on exposure to 50–100% FAS. Heavy metal reduction was in the order Pb>Cd>Zn>Fe>according to accumulation trends. The plant exhibited good biomass growth on exposure to 25–50% FAS, but this decreased at>100% FAS. Heavy metal concentrations in P. hysterophorus after 90 days of the experiment were in the order Fe>Zn>Cu>Pb>Cd>Ni. Parthenium hysterophorus was suitable for translocating Fe, Zn and Cu based on translocation factors (TF=1.5, 1.3 and 1.05), but was more efficient for Pb, Ni and Cd (TF=8.5, 4.3 and 3.3). Plant uptake of Pb, Ni and Cd was high, whereas translocation of Fe, Zn and Cu was poor. These results indicated that P. hysterophorus can efficiently reduce heavy metal pollution in soil.     

Effects of two root-secreted phenolic compounds from a subalpine coniferous species on soil enzyme activity and microbial biomass

In order to evaluate the ecological consequences and potential mechanisms of specific C compounds on soil microbial processes under climate warming, we injected solutions of two modelled root exudates, 2,6-di-tert-butyl-4-methylphenol (BHT) and 1,2-benzenedicarboxylic acid, dibutyl ester (DBP), respectively, into soil at two concentrations (20 and 1000?µg?g^?1 soil). For all treatments, soils amended with the two phenolic compounds were incubated at two temperatures (20°C and 30°C) for 30 days. The responses of soil enzyme activity and microbial property to modelled root exudates to some extent depended on temperature regime, exudation component, and addition concentration. For example, the addition of BHT tended to decrease the soil enzyme activities. However, DBP addition generally increased the two metabolic enzyme activities at 30°C, and tended to decrease the two enzyme activities at 20°C, but a significant reduction was observed only at a high concentration at 20°C. The microbial biomass and enzyme activity were generally lower at 30°C compared to those at 20°C, when averaged across all treatment combinations. Taken together, our results indicated that the amounts and quality of liable root-derived C can differentially affect microbial processes, and various environmental changes will greatly complicate root–microbe–soil interactions in forests.