固定化微生物对土壤中苯并芘的降解

研究了3株细菌与3株真菌对土壤中苯并芘（BaP）的降解动态,从中筛选出1株细菌（Bacillus sp.）和1株真菌（Mucor sp.）,并采用吸附法将混合菌固定在改性后蛭石上,研究了固定化混合菌对土壤中BaP的降解效果。结果表明：细菌中芽孢杆菌（Bacillus sp.,SB02）降解率最高,42 d对B[a]P的降解率为33.0%,降解速率也最快,1周可降解12.6%的BaP;真菌中毛霉（Mucor sp.,SF06）降解率最高,42 d对B[a]P的降解率为69.7%;以改性后蛭石为载体用吸附法制得的固定化混合菌,传质性能好,对BaP的降解率42 d可达95.32%,高于游离菌20个百分点。     

对氯酚的生物降解及其污染土壤的生物修复探索

从天津农芭厂混合土壤中分离到一株能够降解对氯酚的黄单胞菌Ｄ－１,在此基础上,研究了该菌株对对氯酚的降解特性和将Ｄ－１菌株加入模拟对氯污染土壤中的生物修复试验。     

1株贫营养好氧反硝化菌的分离鉴定及其脱氮特性

从水库底泥样品中,以硝酸盐为唯一氮源进行驯化、分离筛选出1株能在贫营养及好氧条件下进行高效反硝化的菌株PY8,经过电镜形态学观察、生理生化和16S rDNA序列分析,并基于16SrDNA序列结果,构建了该菌株的系统发育树,最终确定菌株PY8为根瘤菌Rhizobiumsp.。考察了初始pH值、温度、C/N、初始硝酸钠质量浓度、投菌量对菌株PY8硝酸盐还原活性的影响,以及该菌株的异养硝化性能。结果表明,在pH6.0～10.0,温度25～30℃,C/N1.0～9.0,初始硝酸钠质量浓度0.01～0.50g·L-1,投菌量1%～15%时,菌株PY8培养72h后的硝氮去除率可达到95%以上。另外,该菌株具有同时硝化-反硝化作用,在培养过程中氨氮去除率可达到58%左右。实验结果表明,菌株PY8在微污染水体生物脱氮领域中具有很大的应用潜力。     

Improvement of the assimilable organic carbon (AOC) analytical method for reclaimed water

Microbial growth is an issue of concern that may cause hygienic and aesthetic problems during the transportation and usage of reclaimed water. Assimilable organic carbon (AOC) is an important parameter which determines the heterotrophic bacterial growth potential of water. Pseudomonas fluorescens P17 and Spirillum sp. NOX are widely used to measure AOC in drinking water. The AOC values of various reclaimed water samples determined by P17 and NOX were compared with those determined by the new strains isolated from reclaimed water in this study. It showed that the conventional test strains were not suitable for AOC measurement of reclaimed water in certain cases. In addition to P17 and NOX, Stenotrophomonas sp. ZJ2, Pseudomonas saponiphila G3 and Enterobacter sp. G6, were selected as test strains for AOC measurement of reclaimed water. Key aspects of the bioassay including inoculum cell density, incubation temperature, incubation time and the pH of samples were evaluated for the newly selected test strains. Higher inoculum density (10⁴ CFU·mL^-1) and higher incubation temperature (25°C) could reduce the time required for the tests. The AOC results of various collected samples showed the advantages of the method proposed based on those five strains in evaluating the biologic stability of reclaimed water.     

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.     

铜陵新桥矿区土壤中耐Cu微生物的筛选研究

选择安徽铜陵新桥矿区富Cu的污染土壤,充分利用微生物受自然环境重金属胁迫而产生耐性这一特点,进行土壤中耐Cu微生物的筛选研究。实验过程中分别配制三种不同的培养基,细菌、真菌和放线菌培养基。在水浴恒温振荡器中对土壤中耐受重金属的微生物进行驯化,将得到的对Cu2+耐受性最高的液体培养基作为菌源,在琼脂平板培养基上进行划线分离,并将得到的纯菌株在光学显微镜下进行形态观察,菌种经试管斜面富集培养后保存在4℃冰箱中以便后续使用。研究发现,土样中细菌和真菌对Cu的耐受性低,最高耐受质量浓度分别只达到500mg·L-1和1100mg·L-1。而放线菌表现出Cu高耐受性,分离得到的放线菌耐受Cu的质量浓度最高达到10000mg·L-1,初步鉴定该放线菌株为链霉菌属。该放线菌菌种可能同时对Cu有吸附降解特性,具有成为污染土壤生物治理的高效耐受吸附菌种的潜力巨大。     

假单胞菌AEBL3对土壤中呋喃丹的生物降解

从农药污染的农田土壤中分离到一株假单胞菌AEBL3，该菌能够以呋喃丹为唯一的碳源和氮源生长。使用AEBL3作为生物强化剂对模拟呋喃丹污染的土壤环境进行了生物修复试验。结果表明，接种AEBL3能够明显增加土壤中呋喃丹的降解率。降解菌AEBL3在土壤中具有一定的移动性，当从土壤表层加菌时，对0-7cm深土层中的呋喃丹都有很好的降解效果。在各种投加方式的比较中，以投加降解菌原液修复效果最好，缓冲液悬浮的菌细胞次之，砂粒混合物的效果最差。16S rRNA的变性梯度电泳(DGGE)结果揭示了生物修复过程中土壤微生物菌群结构中的动态变化。     

Screening for plants and rhizospheral fungi with bioremediation potency of petroleum-polluted soils in a Tehran oil refinery area

The contamination of soils by toxic and/or hazardous organic pollutants, especially with crude oil, is a widespread problem. This study was conducted in a petroleum-contaminated area in a Tehran oil refinery to find petroleum-resistant plants and their rhizospheral fungal strains with bioremediation potency. The plants growing in the oil-polluted area were collected and determined taxonomically. Root samples of the plant species were collected from a polluted area and fungal strains determined by laboratory methods and taxonomical keys. The growth ability of the isolated fungal strains was studied in media containing 1%–15% crude oil. Results showed that seven plant species were of the highest density in the contaminated area: Alhagi persarum, Hordeum marinum, Peganum harmala, Phragmites australis, Prosopis farcta, Salsola kali, and Senecio glaucus. The root-associated fungi were isolated and showed that the fungal variation in the oil-polluted area is higher than that in a non-polluted area. The growth assay of isolated fungal strains showed that all studied fungal strains were able to form colonies at the applied concentrations but Alternaria sp. and Rhizopus sp. were the most resistant ones. Some plants were resistant to oil pollution, which also had positive effects on the fungal strains.     

Strengthening effects of ammonia nitrogen on the harmless biological treatment of oily sludge

To improve the efficiency of oil degradation and strengthen the harmless treatment of oily sludge, three dominant strains identified as Chryseomicrobium sp. YL2, Gordonia sp. YL3 and Acinetobacter sp. YL5 were isolated from soil near a refinery, and the effects on the bioremediation of the oily sludge from the refinery were investigated. The results showed that the efficiency of oil degradation increased by 31.5% compared with the control when the dominant strains were added to the treatment of oily sludge. Furthermore, the dominant strains could use oil as a carbon source for heterotrophic nitrification–aerobic denitrification. The addition of ammonia nitrogen resulted in a large number of remaining microbes and heightened dehydrogenase activity in the oily sludge, further accelerating oil degradation, mainly for C₁₁ to C₂₅ saturated hydrocarbons, and the oil degradation efficiency increased by 40.8%. After 120 days of bioremediation, the biotoxicity of oily sludge, which was expressed by the equivalent phenol concentration, decreased by 40.0% compared with that of the control, indicating that the addition of ammonia nitrogen enhanced the biodegradation of oil. This method can be used to strengthen the harmless treatment of oily sludge in practical engineering applications.     

Bioremediation of oily sludge-contaminated soil by stimulating indigenous microbes

In situ bioremediation of oily sludge-contaminated soil by biostimulation of indigenous microbes through adding manure was conducted at the Shengli oilfield in northern China. After bioremediation for 360 days, total petroleum hydrocarbon (TPH) content was reduced by 58.2% in the treated plots compared with only 15.6% in the control plot. Moreover, bioremediation significantly improved the physicochemical properties of the soil in the treated plot. Soil microbial counts and community-level physiological profiling were also examined. Manure addition increased TPH degraders and polycyclic aromatic hydrocarbon (PAH) degraders in the contaminated soil by one to two orders of magnitude. The activity and biodiversity of soil microbial communities also increased markedly in the treated plot compared with that of the control. Finally, biotoxicity was used to evaluate the soils and a sharp increase in the EC50 of the soil after bioremediation was observed, indicating that bioremediation had reduced the toxicity of the soil.     

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.     

应用脂肪酸甲酯淋洗去除土壤中多环芳烃

针对煤气厂土壤等高浓度多环芳烃污染土壤修复困难的现实,采用易生物降解的新型淋洗剂脂肪酸甲酯淋洗修复高浓度多环芳烃污染的土壤,同时进行了以甲醇、植物油(大豆油)作为淋洗剂的淋洗实验,比较不同淋洗剂的淋洗效果.结果证明脂肪酸甲酯对人工模拟污染土壤中蒽、荧蒽、芘、苯并(a)芘的去除率可以达到80%—95%,对煤气厂土壤中多环芳烃的去除效果也非常明显,总多环芳烃的去除率达到41%.脂肪酸甲酯的淋洗效果要优于其它两种淋洗剂.     

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.     

Pollution and biodegradation of hexabromocyclododecanes: A review

• Bioremediation is the most cost-effective approach for degradation of HBCDs. • Bacteria or bacterial consortia are used in the cases of bio-augmentation. • Microbes combined with phytoremediation increase the remediation efficiency. Hexabromocyclododecanes (HBCDs) are the most common brominated flame-retardants after polybrominated diphenyl ethers. HBCDs can induce cancer by causing inappropriate antidiuretic hormone syndrome. Environmental contamination with HBCDs has been detected globally, with concentrations ranging from ng to mg. Methods to degrade HBCDs include physicochemical methods, bioremediation, and phytoremediation. The photodegradation of HBCDs using simulated sunlight or ultraviolet lamps, or chemical catalysts are inefficient and expensive, as is physicochemical degradation. Consequently, bioremediation is considered as the most cost-effective and clean approach. To date, five bacterial strains capable of degrading HBCDs have been isolated and identified: Pseudomonas sp. HB01, Bacillus sp. HBCD-sjtu, Achromobacter sp. HBCD-1, Achromobacter sp. HBCD-2, and Pseudomonas aeruginosa HS9. The molecular mechanisms of biodegradation of HBCDs are discussed in this review. New microbial resources should be explored to increase the resource library in order to identify more HBCD-degrading microbes and functional genes. Synthetic biology methods may be exploited to accelerate the biodegradation capability of existing bacteria, including modification of the degrading strains or functional enzymes, and artificial construction of the degradation microflora. The most potentially useful method is combining micro-degradation with physicochemical methods and phytoremediation. For example, exogenous microorganisms might be used to stimulate the adsorption capability of plants for HBCDs, or to utilize an interaction between exogenous microorganisms and rhizosphere microorganisms to form a new rhizosphere microbial community to enhance the biodegradation and absorption of HBCDs.     

Sol-gel immobilisation of methyl parathion degrading bacteria isolated from agricultural areas of Pakistan

The following areas are discussed in this paper: immobilisation of bacterial consortium in sol-gel; methyl parathion degradation and bioremediation applications; evaluation of indigenous bacterial isolates of contaminated soils. Bacterial strains were isolated from agricultural areas of Pakistan which were contaminated with methyl parathion. A bacterial consortium of seven (out of 64) Enterobacteriaceae isolates including Citrobacter, Enterobacter and Proteus vulgaris capable of degrading methyl parathion (enzyme activity ranging 410–675 mU mL^?1 for individual isolates and 982 mU/mL for consortium) was selected and subsequently immobilised in tetraethyl orthosilicate (TEOS) and sodium-silicate-based sol-gel matrices. Cell viability of suspended and immobilised bacterial consortium was monitored using a minimal salt medium supplemented with methyl parathion. The results indicate that sol-gel immobilisation can be helpful to increase the shelf life of methyl parathion degrading bacterial strains along with preservation of biological activity for bioremediation applications in field.     

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.     

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.     

Impacts of crude oil on the germination and growth of cress seeds (Lepidium sp.) after bioremediation of agricultural soil polluted with crude petroleum using "adapted" Pseudomonas putida

The impacts of crude oil on the germination, growth and morphology of cress seeds (Lepidium sp.) after bioremediation of agricultural soil polluted with crude petroleum using "adapted" Pseudomonas putida (PP) were examined for 15 days. At day 15 there was 100% germination in the untreated control samples, the mean height of the seedlings was 75.8 +/- 2.6 mm and all appeared to have grown morphologically normal. In the experimental samples treated with oil and PP inoculation, there was 98% germination and the seedlings reached a height of 63.8 +/- 6.9 mm; again, morphologically the seedlings appeared normal. However, in the control samples treated with oil but without PP inoculation, there was 31-38% germination and seedling heights of 34.2 +/- 11.4-42.3 +/- 8.5 mm with abnormal morphology. Treatment of oil-impacted agricultural soil with PP as a bioremediation agent does produce soil which is capable of growing larger and healthier plants than where bioremediation has not taken place.     

甲基对硫磷降解菌DLL-1的分离、鉴定及降解性研究

从长期施用甲基对硫磷的污染土壤中分离到一株能以甲基对硫磷为唯一碳源生长且能将其完全矿化的细菌 Ｄ Ｌ Ｌ－ １ ,经鉴定,为邻单胞菌（ Ｐｌｅｓｉｏｍｏｎａｓ ｓｐ ．） ．该菌株３ ｈ 内对５０ ｍｇ／ Ｌ甲基对硫磷的降解率为９３ ％ ,２４ｈ 内对５０ ｍｇ／ Ｌ甲基对硫磷的降解率为９５ ％ 以上．在葡萄糖铵盐培养基中, Ｄ Ｌ Ｌ１ 对甲基对硫磷的耐受浓度和降解速度均增大．降解曲线表明延滞期内,菌体依靠上一生长阶段分泌的酶类对甲基对硫磷进行降解,一旦菌体开始生长,则检测不到中间代谢产物对硝基苯酚的存在．生长情况和粗酶液试验均显示 Ｄ Ｌ１ 优良的降解性状