添加不同营养助剂对石油污染土壤生物修复的影响

实验室条件下研究NPK复合肥、诺沃肥和腐殖酸三种物质不同配比添加对石油污染土壤的生物修复效果.在60d的修复实验中,定期取样测定土壤含油量、总异养菌数、石油烃降解菌数和脱氢酶活性,并采用PCR-DGGE技术研究修复过程中微生物多样性变化.结果表明,NPK肥的添加和NPK肥-诺沃肥-腐殖酸复合添加能够提高土壤中微生物的数量、微生物多样性和脱氢酶活性.在含油量为84600mg.kg-1的土壤中,添加营养助剂的处理60d后石油烃降解率为31.3%—39.5%,不添加营养助剂的石油烃降解率仅为3.5%.NPK肥-诺沃肥-腐殖酸复合添加对石油烃的降解率要高于NPK肥的单独添加(高8%),其原因可能是诺沃肥-腐殖酸能够有效提高土著微生物的活性,增加微生物的多样性,增强石油烃的降解.     

Sulphur-oxidising bacteria isolated from deep caves improve the removal of arsenic from contaminated harbour sediments

Acidophilic S-oxidising bacteria isolated from sulphur-rich deep caves (Frasassi, Italy), characterised by relatively low temperature, were tested for their ability to mobilise (semi-)metals from contaminated sediments. Sediment samples from two commercial Italian seaports were used to set up bioleaching experiments. The effect of different growth substrates was also investigated. Our experiments revealed that S-oxidising bacteria isolated from Frasassi caves have a high potential to remove As from contaminated marine sediments, as never reported before. Although As solubilisation efficiency was quite low (i.e. about 30%), only a small amount of As was associated with non-residual fractions of the sediment. On the contrary, the solubilisation efficiencies of Zn and Ni (20% and 10%, respectively) were lower than those previously obtained by the use of other acidophilic bacteria and mainly influenced by the experimental conditions rather than by the presence of the S-oxidising bacteria. Results presented here open new perspectives in bioleaching applications for the remediation of contaminated sediments. Indeed, microbial strains adapted to relatively low-temperature environments could improve sediment bioleaching at temperature regimes where mesophilic and thermophilic strains are not favoured. Such strains could be exploited for developing selective bioremediation procedures for sediments contaminated with As, to be applied in multistep biotreatment processes.     

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.     

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。     

Alkyl polyglucosides as cell surface modification factors: influence of the alkyl chain length

The use of surfactants in a bioremediation process is aimed at increasing the efficiency of the removal of hydrophobic contaminants from the environment. The subjects of the study were three alkyl polyglucosides (APGs) of different alkyl chain length: Glucopon 215 CS UP, Glucopon 600 CS UP, and Glucopon 650 EC. The impact of these surfactants on the surface properties of the test strain Pseudomonas fluorescens ATCC 14700, as well as on the biodegradation of diesel was tested. It was observed, that the length of alkyl chain has an influence on the cell surface properties. The modification of the cell surface hydrophobicity and the electrokinetic potential on the bacteria cells is dependent on the structure of Glucopon molecules. The elongation of alkyl chains in surfactant molecule caused an increase of the hydrophobic properties and a reduction of the electrokinetic potential on the bacteria cells. Moreover, the use of APGs below critical micelle concentration caused an increase of diesel oil biodegradation, especially in the case of Glucopons with longer alkyl chain (Glucopon 600 CS UP and 650 EC). The better diesel oil removal by tested strain after surfactant addition was correlated with the hydrophobic properties of bacteria strain.     

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.     

垃圾堆肥微生物接种实验

从２２个垃圾堆肥、畜粪、土壤等样品中分离获得纤维分解菌１９８株，选其中２株生长快、粗纤维分解能力强的菌株制成菌剂，以０．０５％～０．１％的接种量加入二次发酵的垃圾堆肥中。结果，接菌堆肥比不接菌堆肥升温快且高，高温维持时间长，真菌和纤维分解菌数量多，腐植质含量提高２１％～２６％。肥效试验证明，施接菌堆肥比施不接菌堆肥的可使青莱增产９．９％。     

产表面活性剂降解石油菌株产物性质及降解性能研究

微生物是组成生态系统的重要成员,在污染物去除中发挥着重要作用,是生物修复中的主力,然而在石油污染修复过程中,石油烃的疏水性会限制微生物对石油的降解,但一些微生物的细胞代谢物即生物表面活性剂,它是微生物在一定条件下代谢分泌产生的具有一定表面活性,集亲水基和疏水基结构于一分子的两亲性化合物,可以促进油的乳化,提高油的分散程度,增大菌株和油珠的接触机会,促进对石油烃的吸收和降解。在实验室分离得到了7株产表面活性剂石油降解菌株,经分子鉴定可知菌1和菌2都为粘质沙雷氏菌Serratia marcescens,菌3为居植物柔武氏菌Raoultella planticola,菌4,菌6和菌7都为克雷伯氏菌Klebsiella variicola,菌5为蜡状芽孢杆菌Bacillus cereus。主要研究了它们的生长与表面活性剂物质分泌状况的关系,发现随着时间增加,OD值随之增大,表面张力呈现下降趋势;并对菌株产物进行提取和薄层层析,离子型分析和红外光谱分析,初步判断其产物均为阴离子糖脂类;通过pH,初始油质量浓度,接种量和盐度4个单因素的变化研究菌1粘质沙雷氏菌,菌3居植物柔武士菌,菌5蜡状芽孢杆菌和菌6克雷伯氏菌对石油类物质降解能力,发现菌3居植物柔武氏菌和菌5蜡状芽孢杆菌降解性能较好;通过响应曲面法优化蜡状芽孢杆菌的降解条件,得出其最佳降解条件为pH为5.02,油质量浓度为3 g·L-1,接种量为1199.98μL,盐度为0.5 g·L-1时,在此条件下,菌株对石油的降解率为66.94%。     

Biodegradation kinetics of cymoxanil in aquatic system

A potential method to detoxify pesticides in aquatic system is using bioremediation. In this study, four microorganisms (Pseudomonas sp (EB11), Streptomyces sp. (EB12), Aspergillus niger (EB13) and Trichoderma viride (EB14) were isolated from cucumber leaves previously treated with cymoxanil using enrichment technique. These strains were evaluated for their potential to detoxify cymoxanil in aquatic system at the concentration level of 5×10^?4M. The effect of pH and temperature on the growth ability of the tested strains was also investigated by measuring the intracellular protein and mycelia dry weight for bacterial and fungal strains, respectively. Moreover, the remaining toxicity of cymoxanil after 28 days of incubation with tested strains was evaluated to confirm the complete removal of any toxic materials (cymoxanil and its metabolites). The results showed that the optimum pH for the growth of cymoxanil degrading strains (bacteria and fungi) was 7. A temperature of 30°C appears to be the optimum for the growth of either fungal or bacterial strains. Pseudomonas sp. (EB11) was the most effective strain in cymoxanil degradation followed Streptomyces sp (EB12), Trichoderma viride (EB14) and Aspergillus niger (EB13), with half-lives of 4.33, 9.5, 17.3 and 24.7 days, respectively. The degradation of cymoxanil by bacterial strains was much faster than fungal one. There is no remaining toxicity of cymoxanil detected in aqueous media previously treated with Pseudomonas sp. (EB11) for 28 days. The results suggest that bioremediation by Pseudomonas sp. (EB11) are promising for the detoxification of cymoxanil in aqueous media.     

光合细菌PSB07-15对辣椒及土壤中甲氰菊酯残留的生物修复

利用光合细菌菌株PSB07-15对辣椒促生作用及对植株与土壤中甲氰菊酯残留的生物修复进行了研究,为光合细菌生物修复菊酯类农药残留的实际应用提供科学依据。结果表明：菌株PSB07-15菌液用ddH2O稀释100倍液、200倍液浸种处理后能够显著提高辣椒种子发芽率;田间小区试验结果表明,菌株PSB07-15施用1 875、3 750、7 500 mL.hm-2,辣椒鲜质量分别增加15.12%、21.68%、14.79%;菌株PSB07-15能够有效降解辣椒和土壤中甲氰菊酯残留（辣椒中大于47.20%,土壤中大于50.73%）。     

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.     

Inorganic nutrient utilisation by "adapted" Pseudomonas putida strain used in the bioremediation of agricultural soil polluted with crude petroleum

Garden soil samples polluted with crude petroleum were bioremediated by inorganic nutrient monitoring with appropriate adjustment and inoculation with crude oil-adapted strain of Pseudomonasputida (PP) isolated from oil-impacted soils. Soil samples without PP inoculation served as the control samples to compare the abilities of the native soil microflora with the adapted PP strain in biodegrading crude oil pollutant. In the experimental samples, oil concentration and all the inorganic nutrient sources tested decreased more rapidly with a proportional increase in the population densities of both PP and the native soil microflora than were observed in the control samples. This trend was particularly strong for PO4(3-) and NO3- which eventually became limiting both in all the experimental samples and in some control samples. Inoculation of crude oil-impacted agricultural soils by oil -adapted PP strain with nutrient monitoring and adjustment can be effective as bioremediation methods of agricultural land upon pollution with petroleum or petroleum products.     

In situ enhanced bioremediation of dichlorvos by a phyllosphere Flavobacterium strain

A bacterium capable of degrading dichlorvos was isolated from the rape phyllosphere and designated YD4. The strain was identified as Flavobacterium sp., based on its phenotypic features and 16S rRNA gene sequence. Strain YD4 was able to utilize dichlorvos as the sole source of phosphorus. In situ enhanced bioremediation of dichlorvos by YD4 was hereafter studied. Chlorpyrifos and phoxim could also be degraded by this strain as the sole phosphorus source. A higher degradation rate of dichlorvos was observed after spraying YD4 onto the surface of rape leaves when compared to the sterilized-YD4 and water-treated samples. The results indicated that pesticide-degrading epiphytic bacterium could become a new way for in situ phyllosphere bioremediation where the hostile niche is unsuitable for other pesticide-degrading bacteria isolated from soil and water.     

转基因毛白杨外源基因转移的环境风险分析

以8年生转Bt基因败育毛白杨雄株为实验材料,分离其体内、体外、根际土壤、蛀干桑天牛(Apripona germari Hope)排泄物中可培养的细菌和真菌,利用特异引物对获得的微生物进行转Bt基因败育毛白杨雄株外源基因PCR检测.实验发现,381株次细菌中有6株PCR检测阳性;307株次真菌中有4株PCR检测阳性.     

Impact of deltamethrin on the endophytic fungal community of a Chinese cabbage,Brassica chinensis

Endophytic fungi, being closely associated with their host plants, are an important part of the ecosystem and food web. In modern agriculture, pesticides are widely used. The impact of pesticides on the endophytic fungal community of crops remains poorly understood. In this paper, Chinese cabbage Brassica chinensis, a popular leafy vegetable consumed worldwide, was selected for evaluating the potential effects of a pesticide, deltamethrin, on the endophytic fungal community. By culture method, 195 endophytic fungal strains classified into 39 taxa and 186 strains classified into 30 taxa, respectively, were isolated from the pesticide-treated and untreated B. chinensis samples. Sirodesmium spp. were the predominant endophytic fungi from both samples. The colonisation rates of endophytic fungi of the treated and the untreated samples were not significantly different, as determined by one-way analysis of variance (p?>?.05). The Shannon diversity indexes (H′) were close, being 2.216 and 2.152, respectively. However, the endophyte compositions of the treated and the untreated samples were significantly different (p?B. chinensis.     

Stimulation of soil microbial activity by essential oils

Summary. Although essential oils are well known antimicrobial agents, some microorganisms are activated by them and can use them as a carbon and energy source; this is the case for soil bacteria from Mediterranean ecosystems. We examined the assumption that soil microorganisms when offered with an essential oil, to which they had been previously exposed, would respond faster making immediate use of the newly added substrate. Origanum vulgare subsp. hirtum, Rosmarinus officinalis, Mentha spicata, and Coridothymus capitatus plants were collected and their essential oils isolated. Soil samples from the upper surface layer, beneath these aromatic plants, were also collected. All possible combinations of essential oils and soil samples were examined as well as the effect of the oil of R. officinalis and the non-indigenous, Lavandula angustifolia, on soil samples collected from cultivated fields. Soil respiration was used as a measure of the microbial activity. Oils (0.1 ml) were repeatedly added to the soil samples (150 g) and CO₂ release was measured every seven days. Essential oils differed in their chemical composition. In spite of that, they activated respiration of the different soil samples, even of those not previously exposed to essential oils, to a comparable degree. These results suggest that essential oils are used as a carbon and energy source by rather ubiquitously occurring soil microorganisms and provide evidence that they would not accumulate in the soil, if environmental conditions favour growth of these microorganisms.     

Pyruvate-utilizing bacteria as potential contributors to the food web in the Arabian Gulf

Water samples from the Kuwaiti coasts of the Arabian Gulf were used for counting and isolating bacteria capable of growth on low molecular weight organic compounds known to be released by picocyanobacteria. The compounds tested were potassium acetate, sodium pyruvate, fumaric acid, succinic acid, sodium citrate and glycerol. For comparison, the bacterial numbers on glucose (a conventional carbon source) and Tween 80 and crude oil (unconventional carbon sources), as sole sources of carbon and energy were also determined. Sodium pyruvate was, in most cases, the carbon and energy source most commonly utilized by the cultivable surface water bacteria. The most common cultivable bacterial genera on the test carbon sources were Pseudoalteromonas, Vibrio, Cobetia and Roseobacter. Less common genera were Rhodococcus, Pseudomonas and Bacillus. Quantification of heterotrophic bacteria associated with cultures of local picocyanobacterial strains, originally isolated from the Gulf surface water, also revealed that the carbon source most commonly utilized by cultivable bacteria was sodium pyruvate. However, a different bacterial composition was identified, with Alcanivorax, Stappia and Marinobacter as the major heterotrophic genera. All heterotrophic bacteria that grew on sodium pyruvate could also grow on β-alanine, as sole sources of carbon and energy. From this study we suggest that the Arabian Gulf bacteria utilizing picocyanobacterial low molecular weight compounds, particularly pyruvate may potentially contribute to the food web in this aquatic system. The experiments comply with the current laws of Kuwait.     

The effects of preen oils and soiling on the UV–visible reflectance of carotenoid-pigmented feathers

Plumage coloration, particularly when carotenoid-based, is important in social signaling in birds. Although feather color is a relatively stable trait, individuals may modify it with “cosmetic” substances such as preen oils. In addition, dirt accumulation may influence plumage coloration and further affect signal perception by receivers. Here, we analyze the separate potential effects of preen oils and soil accumulation on the reflectance properties of carotenoid-pigmented feathers across the visual range of most bird species, which includes the ultraviolet (UV). Using the yellow portion of tail feathers of Bohemian waxwings (Bombycilla garrulus), we performed two separate experiments where: (a) preen oils and/or soil were removed, or (b) preen oils (from black-billed magpies Pica pica or eagle owls Bubo bubo) were added. Preen oil addition reduced brightness but increased UV hue and yellow chroma. UV chroma was reduced by the addition of magpie (but not owl) preen oil. Soil accumulation had little effect on plumage reflectance in the UV range but significantly reduced yellow chroma. According to models of avian vision, both of these effects are detectable by birds and biologically meaningful when compared with natural variation between the sexes and age classes. We conclude that preen oil and soil accumulation can significantly affect the UV–visible reflectance of carotenoid-based plumages. As such traits typically advertise individual quality, preening and soiling have the potential to modify the information content of carotenoid-based plumage traits and how these signals are perceived by receivers.     

Evidence for n-alkane consumption and oxidation by filamentous cyanobacteria from oil-contaminated coasts of the Arabian Gulf

Microcoleus chthonoplastes and Phormidium corium were isolated from microbial mats covering all sediments along the Arabian Gulf coasts. These isolates could consume and oxidise n-alkanes. The establishment of axenic cultures faced the problem that with progressive axenity the cyanobacterial growth seemed to cease. The associated organotrophic bacteria, Rhodococcus rhodochrous, Arthrobacter nicotianae, Pseudomonas sp. and Bacillus sp., could utilize n-alkanes. The total number of these organotrophs was about 2×10⁶ cells g⁻¹ fresh culture, and R. rhodochrous was the most dominant. In order to test the potential of cyanobacteria for n-alkane consumption, experiments were constructed to rule out the role of the associated organotrophic bacteria. Aliquots, 0.5 g fresh cyanobacterial samples, each containing about 1×10⁶ organotrophic bacterial cells (≡0.001 mg fresh bacteria) were incubated in inorganic medium aliquots supplied with an n-alkane. The same was repeated using 1.0×10⁶ cells each of the four organotrophic bacteria instead of the cyanobacterial samples. The nonaxenic cyanobacterial samples consumed up to 60% of the available alkane, whereas no detectable consumption was measured in any of the pure organotrophic bacterial cultures. For all organotrophic bacteria, the numbers had to be increased ten-thousand times in order that detectable alkane consumption might become measurable. The fatty acids resulting from the n-alkane oxidation were found incorporated in cell lipid classes characteristic of cyanobacteria, namely in galactolipids and sulfolipids. These results may imply that the two test cyanobacteria contribute directly to n- alkane uptake and oxidation. Received: 6 May 1997 / Accepted: 2 October 1997     

Effects of Arbuscular mycorrhizal fungi isolated from white clovers (Trifolium repens L) on soil bacteria and fungi

White clover potted experiments were performed to investigate the effects of seven indigenous arbuscular mycorrhizal fungi (AMF) communities isolated from different test plots subjected to long-term fertilisation on soil enzyme activities, number of soil bacteria and fungi. The results showed that the inoculation of arbuscular mycorrhizal fungi communities increased the mycorrhizal infection rate of the plants and promoted the growth of plants. The Mnp treatment was most effective. The shoot biomass, root biomass, potassium and nitrogen uptake of the white clover in Mnp treatment group were increased by 61.54%, 84.00%, 62.50% and 46.71% respectively, compared with those in non-inoculation treatment. The inoculation of AMF communities had little effect on the number of bacteria in the soil, but significantly increased the number of soil fungi. Mnk treatment group had the highest number of fungi in the soil, which was 9.91 times that of the non-inoculation treatment group. The catalase and dehydrogenase activities were both significantly improved in Mnp treatment by 28.12% and 205.38% respectively, compared with those of the control treatment (-M). The urease, invertase and cellulase activities reached the highest levels in the Mck treatment; they were increased by 142.79%, 41.17% and 77.62% respectively, compared with those of the control treatment. Pearson correlation analysis showed that the soil enzyme activity was not correlated with the mycorrhizal infection rate, but correlated with the spore number of the AMF community. The impact of AMF community on soil quality is important for us to understand the function of the ecosystems. Relevant study provides important guidance for maintaining the balance of the soil-plant system and the development of sustainable agriculture.