鼠李糖脂在铜绿假单胞菌NY3代谢烃过程中的生理作用

为了解鼠李糖脂在菌代谢烃过程中所起的具体作用,对NY3菌体生理生化指标进行测定,研究了NY3菌降解烃过程中,鼠李糖脂促进烃降解与细胞生理特性变化规律的相关性。结果表明,投加60、100和140 mg/L鼠李糖脂,能使正十六烷降解率分别提高23.7%、48.3%和32.2%,且产酸明显。鼠李糖脂能加快疏水性有机物的传质速度,与未加鼠李糖脂相比,鼠李糖脂浓度为100和140 mg/L时,菌细胞内积聚的正十六烷量增加12.5%和37.69%。鼠李糖脂存在下,NY3菌细胞内正十六烷吸收是一个积累和消耗相互交替的动态过程。鼠李糖脂使菌体细胞中疏水性脂类的相对含量和细胞表面的负电荷明显增加,但使所生长的细胞内脂多糖含量明显减少,生长过程投加鼠李糖脂浓度为60、100、140和200mg/L时,细胞中脂多糖含量分别减少45.65%、58.13%、45.61%、和59.44%。因此,鼠李糖脂能加快NY3菌对十六烷的降解,同时对代谢过程中菌体的生理生化特性有明显的改变。     

鼠李糖脂浓度对铜绿假单胞菌NY3烃代谢过程中细胞生理特性影响作用

为了研究表面活性剂鼠李糖脂(Rha)促进铜绿假单胞菌NY3烃降解与该菌细胞生理特性变化规律的相关性,在实验室可控条件下,研究了投加Rha后对该菌在整个培养过程中生长情况、正十六烷降解率、胞内脂多糖含量、胞内外蛋白含量等生理特性的影响。结果表明,NY3菌对正十六烷的降解率不受限于该鼠李糖脂临界胶束浓度(CMC=35 mg·L~(-1)),即在投加高于CMC浓度时同样可以提高正十六烷的降解率,并增大体系生物量和积累大量酸性物质(多为烃的氧化产物),该酸性物质能被NY3菌利用并快速生长。投加Rha可增加细胞内脂多糖的含量,并与体系中被去除的烃类物质的量成正比;同时能促进胞内蛋白在短时间内增加并趋于稳定,而降解体系中胞外蛋白同样对于正十六烷的降解有积极作用。因此,投加Rha不仅能提高对疏水性有机物的增溶性,同时对代谢过程中菌体的生理生化特性有明显的改变,从而提高NY3菌对烃类物质的降解率。     

鼠李糖脂对微生物菌剂降解石油的影响

以石油烃降解微生物菌剂和铜绿假单胞菌株A6为对象，考察不同浓度鼠李糖脂对菌剂细胞表面疏水性、原油降解性能和微生物生长的影响，采用GC-MS分析石油中正构烷烃组分的降解情况。结果表明，低浓度鼠李糖脂就可提高菌剂细胞的表面疏水性和原油降解效果。以250mg／L添加组最明显，第7天疏水性达最高，为58．6％，比对照组提高约26．2％；降解第15天原油降解率达71．6％，分别比对照组和TW20组提高16％和13．3％。GC-MS结果显示鼠李糖脂对高碳数烷烃的降解作用大于低碳数烷烃，正二十三烷和正三十三烷的降解率分别较对照提高了21．5％和33．7％。菌剂对奇数碳烷烃的降解效果优于偶数碳烷烃。鼠李糖脂分别使菌剂中细菌、放线菌和霉菌的最大生物量提高了5．7、2．4和1．8倍。鼠李糖脂对微生物细胞疏水性和生物量的提高与石油降解效果正相关。     

二鼠李糖脂对白腐菌降解稻草中木质纤维素的影响

通过固态发酵方式采用白腐菌对稻草中木质纤维素进行降解,并研究了生物表面活性剂二鼠李糖脂对该降解过程的影响。结果表明,不同浓度的二鼠李糖脂能在不同程度上提高降解过程水溶性有机碳的含量,添加0.007%和0.021%二鼠李糖脂的实验组最高TOC(total organic carbon)浓度较对照组分别提高了83.6%和54.5%,这有利于黄孢原毛平革菌的生长,且延缓了菌体的衰退。添加临界胶束浓度0.007%和0.021%浓度的二鼠李糖脂可使LiP(lignin peroxidase)酶活分别提高85.7%和41.2%,二鼠李糖脂对MnP(manganese peroxidase)酶活没有显著影响。生物表面活性剂的介入促进了白腐菌对稻草中木质素的降解,添加0.007%二鼠李糖脂可使木质素降解率提高54%。     

嗜油菌NY3对蜡质油污泥无害化处理

蜡质油污泥(WOS)成分复杂,含大量老化原油、蜡质油、沥青质及其他化学药剂,属危险性固体废弃物,大量排放已成为石化企业可持续发展的障碍.研究利用铜绿假单胞菌NY3处理高浓度WOS的条件,结果表明,WOS∶木屑(m/m)(d =0.30 mm) 7∶1并结合热熔分散为最佳油污泥预处理条件;液相降解体系含油量为24.5 g/L时,6d内,油泥中C20以下的正构烷烃完全被去除,C21～C34和C35～C38大分子量正构烷烃去除率分别在91％ ～99％和78％ ～ 89％之间.添加鼠李糖脂能明显提高长链烃和多环芳烃的降解率.6d内210 mg/L鼠李糖脂使C29 ～ C38的降解率提高17.27％ ～36.65％,芘、菲分别提高13.67％和16.12％.添加葡萄糖和Fe2+的条件下,NY3菌可将油污泥转化成为糖蛋白,产量达28 g/L.     

利用不同组分原油逐级驯化筛选高效石油烃降解混菌

利用不同组分原油逐级驯化的方法对克拉玛依油田的石油污染土样进行石油烃降解混菌的富集驯化,得到一组对稀油和稠油均具有高效降解能力的混菌M3。与采用单一原油驯化方法相比,混菌M3对稀油和稠油的降解率分别提高了12.5%和22%。该混菌具有较强的产表面活性剂的能力,能够使发酵液的表面张力从69.8 mN·m~(-1)降至27.9 mN·m~(-1)。通过混菌M3的生长条件优化实验得出:在温度30℃、pH 7～8、盐度1%、氮源选择尿素的条件下,混菌M3对原油的降解率最高。通过考察混菌M3在污染土壤中对原油的降解效果,发现:在修复期间,土壤脱氢酶呈先升高后降低的趋势;混菌M3可使饱和烃组分增加,并使芳香分、胶质和沥青质组分降低,对重质组分具有较好的降解效果。混菌M3的加入改变了原油性质,促进了土壤中原油的降解,经过56 d修复,土壤中原油降解率达到55.3%。     

鼠李糖脂对不同菌株降解柴油污染物的影响

通过一系列实验分析了鼠李糖脂对柴油污染物生物降解的影响。单菌株柴油降解实验结果表明,在添加生物表面活性剂鼠李糖脂后,各菌株细胞表面疏水性均发生不同程度的增加,并且对柴油的降解率均有所提高。在混合菌的柴油污染物降解实验中,发现当向土壤中添加了200 mg/L鼠李糖脂时,对柴油的降解才有较大的提高;而当添加100 mg/L的鼠李糖脂到水体中时,对柴油的降解就有较大的提高,而当鼠李糖脂浓度提高为200 mg/L时,柴油的降解率却没有进一步明显的提高。这说明鼠李糖脂对柴油降解的影响程度不仅与环境介质有关,还与添加的鼠李糖脂浓度有关。进一步分析表明,添加适当浓度的鼠李糖脂不仅可以提高对柴油的降解率,而且可加速其降解速度,缩短生物修复所需时间。     

石油降解菌的筛选优化及其对油污土壤的修复特性

分别以牛肉膏蛋白胨-布氏哈斯培养基、蓝色凝胶培养基作为初筛和复筛培养基,从石油污染土壤中筛选出2株可产生微生物表面活性剂的石油烃降解菌。并将菌株投加到油污土壤中进行修复研究,考查了不同影响因素对修复效果的影响。研究结果表明,(1)2株菌对中度石油污染土壤有较好的修复效果,向油污土壤中直接投加菌株修复70 d时对石油烃的去除率为52%;(2)向油污土壤中投加降解菌并同时补充氮营养液,修复70 d时对土壤中总石油烃的去除率可达到75%;对土壤中正构烷烃的去除率为66%;(3)与土壤的含水率及土著菌的降解效果相比,向油污土壤中投加降解菌以及补充氮磷营养液是影响石油污染土壤修复效果的关键因素。     

假单胞菌L-01产表面活性剂的性能分析

通过对活性部位测定,表明菌株L-01产的生物表面活性剂主要是胞外型的糖脂类物质;提取生物表面活性剂后,采用红外扫描仪和紫外扫描仪测定其结构并进一步分析表明,该生物表面活性剂为鼠李糖脂。当以柴油为惟一碳源培养菌株L-01时,在生长的对数后期开始产鼠李糖脂,并在稳定期的初期达到最大值。菌株L-01产的鼠李糖脂对柴油具有良好的乳化能力,培养60 h时,其E24值高达95.2%。通常表面活性剂的临界胶束浓度(CMC)是指在水中测得的,而对鼠李糖脂在水中和土壤中的测定其CMC值后,结果发现两者差距很大,在水中的CMC为65 mg/L;而在土壤溶液中时,其CMC值高达185 mg/L,这一点对利用生物表面活性剂于石油污染生物修复具有重要的参考价值,因为添加的表面活性剂在高于CMC值时才能有效地提高生物修复效率。     

固定化微生物修复石油污染土壤影响因素研究

针对石油污染土壤修复,利用实验室已筛选的高效石油降解单菌SM-3,以天然有机材料为载体,吸附法制备固定化微生物。将游离与固定化微生物应用于室内花盆模拟修复石油污染土壤,对C/N/P、微生物投加量、石油含量、氧化剂和表面活性剂设计5因素4水平正交实验,探讨不同修复时期各影响因素的重要性顺序,最佳条件下各菌株的修复效果。结果表明,不同微生物在不同降解时期,各影响因素的重要性会发生变化;经过21 d的修复,固定化单菌SM-3石油降解率为22.77%,修复过程中,接种量是最重要的影响因素,营养元素N、P投加影响较大,表面活性剂和氧化剂影响次之。     

Integration of biological responses from a suite of bioassays for the Venice Lagoon (Italy) through sediment toxicity index - Part A: Development and comparison of two methodological approaches

Marine and coastal quality assessment, based on test batteries involving a wide array of endpoints, organisms and test matrices, needs for setting up toxicity indices that integrate multiple toxicological measures for decision-making processes and that classify the continuous toxicity response into discrete categories according to the European Water Framework Directive.Two toxicity indices were developed for the lagoon environment such as the Venice Lagoon. Stepwise procedure included: the construction of a database that identified test-matrix pairs (indicators); the selection of a minimum number of ecotoxicological indicators, called toxicological core metrics (CMs-tox) on the basis of specific criteria; the development of toxicity scores for each CM-tox; the integration of the CMs-tox into two indices, the Toxicity Effect Index (TEI), based on the transformation of Toxic Unit (TU) data that were integrated as logarithmic sum, and the Weighted Average Toxicity Index (WATI), starting from toxicity classes integrated as weighted mean. Results from the indices are compared; advantages and drawbacks of both approaches are discussed.     

Comparative bioremediation potential of four rhizospheric microbial species against lindane

Four microbial species (Kocuria rhizophila, Microbacterium resistens, Staphylococcus equorum and Staphylococcus cohnii subspecies urealyticus) were isolated from the rhizospheric zone of selected plants growing in a lindane contaminated environment and acclimatized in lindane spiked media (5-100 μg mL⁻¹). The isolated species were inoculated with soil containing 5, 50 and 100 mg kg⁻¹ of lindane and incubated at room temperature. Soil samples were collected periodically to evaluate the microbial dissipation kinetics, dissipation rate, residual lindane concentration and microbial biomass carbon (MBC). There was a marked difference (p < 0.05) in the MBC content and lindane dissipation rate of microbial isolates cultured in three different lindane concentrations. Further, the dissipation rate tended to decrease with increasing lindane concentrations. After 45 d, the residual lindane concentrations in three different spiked soils were reduced to 0%, 41% and 33%, respectively. Among the four species, S. cohnii subspecies urealyticus exhibited maximum dissipation (41.65 mg kg⁻¹) and can be exploited for the in situ remediation of low to medium level lindane contaminated soils.     

Inhibition kinetics of certain enzymes in the nervous tissue of vector snail Lymnaea acuminata by active molluscicidal components of Sapindus mukorossi and Terminalia chebula

Effect of active molluscicidal components of Sapindus mukorossi and Terminalia chebula on the acetylcholinesterase (AChE), acid and alkaline phosphatase (ACP/ALP) activity in the nervous tissue of freshwater snail Lymnaea acuminata were studied. In vivo and in vitro exposure of saponin (active component of S. mukorossi pericarp) and tannic acid (active component of T. chebula) significantly inhibited the AChE, ACP and ALP activity in the nervous tissue of L. acuminata. The inhibition kinetics of these enzymes indicate that saponin and tannic acid caused competitive and competitive-non-competitive inhibition of AChE, respectively. Saponin also caused competitive and competitive-non-competitive inhibition of ACP and ALP, respectively, whereas tannic acid caused competitive-non-competitive inhibition of ACP and ALP. Thus the inhibition of AChE, ACP and ALP by saponin and tannic acid in the nervous tissue of L. acuminata may be the cause of molluscicidal activity of S. mukorossi and T. chebula.     

Lethal effects of abamectin on the aquatic organisms Daphnia similis, Chironomus xanthus and Danio rerio

Abamectin is used as an acaricide and insecticide for fruits, vegetables and ornamental plants, as well as a parasiticide for animals. One of the major problems of applying pesticides to crops is the likelihood of contaminating aquatic ecosystems by drift or runoff. Therefore, toxicity tests in the laboratory are important tools to predict the effects of chemical substances in aquatic ecosystems. The aim of this study was to assess the potential hazards of abamectin to the freshwater biota and consequently the possible losses of ecological services in contaminated water bodies. For this purpose, we identified the toxicity of abamectin on daphnids, insects and fish. Abamectin was highly toxic, with an EC₅₀ 48 h for Daphnia similis of 5.1 ng L⁻¹, LC₅₀ 96 h for Chironomus xanthus of 2.67 μg L⁻¹ and LC₅₀ 48 h for Danio rerio of 33 μg L⁻¹.     

Direct and indirect effects of ammonia, ammonium and nitrate on phosphatase activity and carbon fluxes from decomposing litter in peatland

Here we investigate the response of soils and litter to 5 years of experimental additions of ammonium (NH₄), nitrate (NO₃), and ammonia (NH₃) to an ombrotrophic peatland. We test the importance of direct (via soil) and indirect (via litter) effects on phosphatase activity and efflux of CO₂. We also determined how species representing different functional types responded to the nitrogen treatments. Our results demonstrate that additions of NO₃, NH₄ and NH₃ all stimulated phosphatase activity but the effects were dependent on species of litter and mechanism (direct or indirect). Deposition of NH₃ had no effect on efflux of CO₂ from Calluna vulgaris litter, despite it showing signs of stress in the field, whereas both NO₃ and NH₄ reduced CO₂ fluxes. Our results show that the collective impacts on peatlands of the three principal forms of nitrogen in atmospheric deposition are a result of differential effects and mechanisms on individual components.     

Suitability of different salt marsh plants for petroleum hydrocarbons remediation

The suitability of the salt-marsh species Halimione portulacoides, Scirpus maritimus, Juncus maritimus and an association of the last two for remediation of petroleum hydrocarbons (PHC) in soil was investigated. An outdoor laboratory experiment (microcosm-scale) was carried out using contaminated soil collected in a refinery, as a complement of another study carried out in the refinery environment (mesocosm-scale). Soil samples with old contamination (mainly crude oil) and with a mixture of the old and recent (turbine oil) contamination were tested. Studies in both micro- and mesocosm-scale provided results coherent in substance. The presence of S. maritimus caused removal of old contamination which was refractory to natural attenuation (after 7 months of exposure, efficiency was 13% when only old contamination was present and 40% when the soil also contained recent contamination). H. portulacoides (only included in the microcosm-scale study) revealed also potentiality for PHC remediation, although with less efficiency than S. maritimus. Degradation of recent contamination was also faster in the presence of plants (after 7 months: 100% in the presence of S. maritimus vs. 63% in its absence). As these species are common in salt marsh areas in Atlantic coast of Europe, it is probable they will be also useful for recovering coast sediments. In contrast, J. maritimus and association did not reveal capability to remove PHC from soil, the presence of J. maritimus inhibiting the capability of S. maritimus.     

Inter- and intra-specific responses to elevated ozone and chamber climate in northern birches

We studied the responses of micropropagated, northern provenances of downy, mountain and silver birches to elevated ozone (O₃) and changing climate using open-top chambers (OTCs). Contrary to our hypothesis, northern birches were sensitive to O₃, i.e. O₃ levels of 31-36 ppb reduced the leaf and root biomasses by −10%, whereas wood biomass was affected to a lesser extent. The warmer and drier OTC climate enhanced growth in general, though there were differences among the species and clones, e.g. in bud burst and biomass production. Inter- and intra-specific responses to O₃ and changing climate relate to traits such as allocation patterns between the above- and belowground parts (i.e. root/shoot ratio), which further relate to nutrient and water economy. Our experiments may have mimicked future conditions quite well, but only long-term field studies can yield the information needed to forecast responses at both tree and ecosystem levels.     

The exposition of a calcareous Mediterranean soil to toxic concentrations of Cr, Cd and Pb produces changes in the microbiota mainly related to differential metal bioavailability

The involvement of the bacterial community of an agricultural Mediterranean calcareous soil in relation to several heavy metals has been studied in microcosms under controlled laboratory conditions. Soil samples were artificially polluted with Cr(VI), Cd(II) and Pb(II) at concentrations ranging from 0.1 to 5000 mg kg⁻¹ and incubated along 28 d. The lowest concentrations with significant effects in soil respirometry were 10 mg kg⁻¹ Cr and 1000 mg kg⁻¹ Cd and Pb. However, only treatments showing more than 40% inhibition of respirometric activity led to significant changes in bacterial composition, as indicated by PCR-DGGE analyses. Presumable Cr- and Cd-resistant bacteria were detected in polluted microcosms, but development of the microbiota was severely impaired at the highest amendments of both metals. Results also showed that bioavailability is an important factor determining the impact of the heavy metals assayed, and even an inverted potential toxicity ranking could be achieved if their soluble fraction is considered instead of the total concentration. Moreover, multiresistant bacteria were isolated from Cr-polluted soil microcosms, some of them showing the capacity to reduce Cr(VI) concentrations between 26% and 84% of the initial value. Potentially useful strains for bioremediation were related to Arthrobacter crystallopoietes, Stenotrophomonas maltophilia and several species of Bacillus.     

Trace element accumulation in woody plants of the Guadiamar Valley, SW Spain: a large-scale phytomanagement case study

Phytomanagement employs vegetation and soil amendments to reduce the environmental risk posed by contaminated sites. We investigated the distribution of trace elements in soils and woody plants from a large phytomanaged site, the Guadiamar Valley (SW Spain), 7 years after a mine spill, which contaminated the area in 1998. At spill-affected sites, topsoils (0-25 cm) had elevated concentrations of As (129 mg kg(-1)), Bi (1.64 mg kg(-1)), Cd (1.44 mg kg(-1)), Cu (115 mg kg(-1)), Pb (210 mg kg(-1)), Sb (13.8 mg kg(-1)), Tl (1.17 mg kg(-1)) and Zn (457 mg kg(-1)). Trace element concentrations in the studied species were, on average, within the normal ranges for higher plants. An exception was white poplar (Populus alba), which accumulated Cd and Zn in leaves up to 3 and 410 mg kg(-1) respectively. We discuss the results with regard to the phytomanagement of trace element contaminated sites.     

Potential re-colonisation by cladocerans of an acidic tropical pond

To predict how re-colonisation of acidified lakes will proceed, at least two approaches are possible: (i) to compare the life history traits of candidate species and determine which one has the highest fitness, and (ii) to simulate a more realistic scenario carrying out experiments with the grouping of the candidate species, so that the intrinsic rate of natural increase of each species is integrated with its sensitivity to low pH and its ability to compete with the other candidate populations. The objective of the present study was to investigate the future re-colonisation of such acidified systems, taking as case-study a tropical pond (Lagoa das Dunas, Camaçari, BA, Brazil) and four species of cladocerans occurring in nearby water bodies (Ceriodaphniacornuta, Ceriodaphniasilvestrii, Latonopsisaustralis and Macrothrix elegans), by comparing the two above mentioned approaches. The second approach included two sets of in situ microcosms experiments, one simulating the re-colonisation by immigrating ephippia, thus using neonates of each species as colonisers, and another simulating the immigration of adults. Both these simulations followed nearly the same trends. The integration of the effects of a higher temperature, a different photoperiod and species competition determined differences in the species densities ranking between the two approaches: life history versus microcosms. The densities of C.cornuta in the microcosms matched the biphasic concentration/response hormetic model, in the simultaneous presence of two increasingly intense stressors (interspecific competition and acidity), with a low-dose stimulation and a high-dose inhibition. The present study provided, thus, a further support to the acceptance of hormesis in ecotoxicology, also at the population level in multispecies experiments.