黑麦草-耳葡萄球菌对水体中Cu的联合修复效应研究

采用水培实验方法模拟不同程度的Cu污染状况,研究了黑麦草与耳葡萄球菌联合修复作用对Cu的去除效果。结果表明,联合修复作用对Cu的去除效果明显优于单一的黑麦草修复,Cu初始浓度在5-30mg.L-1时,联合修复对Cu的去除率均达85%以上。耳葡萄球菌对Cu表现出较强的耐受性和吸附性,Cu浓度为100mg.L-1时其干重积累量达到40.6mg.g-1。耳葡萄球菌的存在降低了黑麦草根系及茎叶中Cu的积累量,联合修复组的黑麦草的生物量和叶绿素含量明显高于单一黑麦草修复组。该菌在一定程度上减轻了黑麦草所受Cu的毒害性,提高了水中Cu的联合修复效果。     

Molecular Monitoring of SRB Community Structure and Dynamics in Batch Experiments to Examine the Applicability of in situ Precipitation of Heavy Metals for Groundwater Remediation (15 pp)

Background, Aims and Scope   Sulfate-reducing bacteria (SRB) are known for their capacity to reduce and precipitate heavy metals (HM) as metal sulfides, offering the opportunity to create an in situ reactive zone for the treatment of heavy metal-contaminated groundwater, a process called in situ metal precipitation (ISMP). The applicability of the ISMP technology first has to be investigated at a laboratory scale before going into an on site application. The evaluation and optimization of the ISMP process is facilitated when physical/chemical analysis techniques are combined with molecular tools that specifically monitor the abundance, diversity and dynamics of the indigenous sulfate reducing microbial community. In this study, batch experiments were conducted in order to investigate the feasibility of ISMP as a groundwater remediation strategy for an industrial site contaminated with elevated levels of Zn, Cd, Co and Ni. Methods   The potential of different types of carbon source/ electron donor (lactate, acetate, methanol, ethanol, Hydrogen Release Compound?, molasses) to stimulate the sulfate reduction and metal precipitation activity of the naturally present (or indigenous) SRB community was explored. In addition, the effect of amending vitamin B12 and yeast extract was evaluated. The ISMP process was monitored by combining analytical analyzes of process parameters (SO42-concentration, heavy metal concentrations, pH, Eh) with molecular tools such as SRB subgroup and genus specific PCR, denaturing gradient gel electrophoresis (DGGE), and phylogenetic analysis of clone sequences, based on either the 16S rRNA or the dsr (dissimilatory sulfite reductase) gene. Results and Discussion   The efficiency of different carbon-sources to stimulate the ISMP process followed the order HRC 〉 molasses 〉 methanol 〉 lactate 〉 ethanol 〉 acetate. Within 10 weeks, the highest sulfate and metal removal efficiencies ranged from 85% to 99%. Addition of yeast extract boosted the ISMP process, whereas vitamin B12 negligibly affected SRB activity. Analysis of the sulfate reducing population by SRB subgroup and genus specific PCR demonstrated that members of the genus Desulfosporosinus dominated in all batch tests, while 16S rDNA DGGE profiles additionally revealed the presence in the microbial communities of non-sulfate reducing bacteria within the family Clostridium and the -proteobacteria. The dsrB-based DGGE profiles allowed us to assess the diversity and dynamics of the sulfate reducing community and added to a better understanding of the effects of different batch conditions on the ISMP process. Remarkably, all dsrB sequences affiliated with the dsrB gene sequence cluster found in Desulfotomaculum, which received their xenologous dsrB gene from the -proteobacteria. Conclusions   The batch experiments, which aimed at stimulating the activities of the indigenous SRB communities, demonstrated that these communities were present and that their activities could be used to obtain efficient in situ precipitation of the contaminating heavy metals. This opens the possibility to test this concept in the future as an on site demonstration as part of the groundwater strategy for the heavy metal contaminated site. Although batch setups are suitable for preliminary feasibility studies for ISMP, they do not reflect the in situ situation where sulfate and heavy metal and metalloid polluted groundwater are supplied continuously. A sulfate reducing strain JG32A was isolated from whose 16S rRNA gene affiliated with the genus Desulfosporosinus, while its dsrB gene sequence clustered with Desulfotomaculum dsrB gene sequences, which received their xenologous dsr genes from -proteobacteria. Therefore we hypothesize that the batch experiments enrich members of the Desulfosporosinus genus that possess a non-orthologous dsrB gene. Recommendation and Perspective   The next step towards an on site pilot test for ISMP will be the setup of a series of column experiments, with process conditions that are selected based on the above mentioned results. This will allow to define optimal ISMP process conditions and to test its long-term efficacy and sustainability before going into an on site bioremediation application. By applying the described molecular tools together with physical-chemical analyzes, it can be investigated whether the same SRB community is enriched and which type of C-source is most effective in promoting and sustaining its growth and sulfate-reduction activity.     

石油污染土壤的生物修复研究

通过田间试验研究了玉米和向日葵两种植物对石油污染土壤的修复作用,考察了外源菌（OX-9）对植物修复的强化和协同效应,对“外源菌一植物”修复效果进行了初步评价。结果表明,在10000mg·k-1污染浓度下,150d玉米、向13葵试验区土壤中石油降解率分别为42．5％和46．4％,较对照区提高了100．5％和118．9％。外源节细菌的施加可使生物修复速度显著加快,150d“DX-9-玉米”和“DX-9-向日葵”试验区石油烃降解率分别达到72．8％和76．4％,较同期单独植物修复的降解率提高了71.3％和64．7％。500d各试验区土壤中石油烃降解率分别为95．5％、96．1％、97．6％和98．9％,土壤中石油烃含量均低于国家标准规定限量（〈500mg·kg-1）;土壤主要理化性质、生物群落分布、呼吸强度及植物不同部位中石油烃的残留量与对照无显著差异。结果表明：玉米、向日葵与节细菌对石油污染土壤的联合生物修复效果显著;经过两年修复,污染土壤恢复健康状态。     

Effects of rhamnolipids on bacterial communities in a dioxin-contaminated soil and the gut of earthworms added to the soil

The biosurfactants rhamnolipids and the “soil ecosystem engineers” earthworms are often used to remediate contaminated soils. However, the effects of rhamnolipids on earthworm intestinal flora and microbial community in soil containing earthworms are not clearly understood. In our study, a 21-d microcosm experiment was carried out to reveal the effects of rhamnolipids on microbial abundance, composition, and metabolism, as well as contaminant degradation capacity. Both rhamnolipids and earthworm...     

玉足海参与凡纳滨对虾的混养效果

在室内条件下进行了玉足海参与凡纳滨对虾的混养实验,分析了单养与混养两种条件下养殖水体营养盐结构以及底质成分的变化,测定了对虾与海参的存活率与生长性能。结果显示,混养海参可以明显改变养殖系统的营养盐结构,可使水体中的磷酸盐和硝酸盐浓度有所升高,同时也可有效地控制系统中氨氮浓度。混养海参也可以大幅度地降低沉积物中有机质和硫化物含量,实验结束时混养组硫化物含量为(7.71±1.33)mg/kg,仅相当于单养组浓度的1/3。混养海参对对虾生长及存活具有明显的促进作用,其中混养组对虾体长特异增长率为(0.69±0.13)%/d,显著优于单养组(0.45±0.06)%/d;混养组对虾成活率可达72.5%±22.9%,显著高于对照组55.0%±17.5%。在混养系统内,对虾不会对海参的生存造成负面影响,海参能够有效地选择摄食和利用沉积物中的营养物质(对食物中有机质的同化率可达36.36%±13.79%),并以较快的速度生长。结果表明,在对虾养殖系统中混养玉足海参具有明显的环境与经济效益。本研究可为我国海水养殖业的可持续发展提供一定的科学依据。     

Well-defined multispecies probiotic and enzyme combination outperforms traditional fermented probiotic applications in an intensive Pacific white shrimp,Litopenaeus vannamei (Boone, 1931), culture system

A series of treatments were designed to evaluate the efficacy of feeding commercial multispecies probiotics feeding with enzymes and fermentation process on the growth parameters and culture environment of Pacific white shrimp, Litopenaeus vannamei (Boone, 1931), in an intensive culture system. Commercial multispecies probiotics and enzymes (PEs) were continuously applied in three different doses, namely (i) 0.2, (ii) 0.4, and (iii) 0.6 mg L⁻¹ and designated as 0.2, 0.4, and 0.6 PE during the first 30 days of intensive culture of Pacific white shrimp, L. vannamei (Boone, 1931). The probiotics were continually applied every alternate day, while the enzymes were added every sixth day throughout the trial period. The PE dose for all treated tanks was increased by 0.2 ppm after 30 days of culture and another 0.2 ppm after day 60. Meanwhile, fermentation technique, which has become the common method applied in Indonesia in the control treatment, was added on the same day with the PE group with increasing dosage, following the same trend with the 0.6 PE group. Results showed that the group receiving 0.6 PE showed higher final biomass, higher mean weight, and protein retention efficiency as well as a lower feed conversion ratio compared with the control treatment. Abundance of Vibrio spp. remained below 10³ cfu mL⁻¹ throughout the trial. Water quality indicators TAN, NO₂ N and NO₃ N peaked in weeks 3–5 and then declined after that until the end of the culture period in all tanks. This decline was significantly faster in PE-treated tanks. Multispecies PEs have potential applications in controlling Vibrio spp., maintaining proper water quality condition, and enhancing the growth of shrimp in intensive culture system.     

参后地土壤中五氯硝基苯微生物修复效应及影响因素研究

从长期使用五氯硝基苯(PCNB)的栽参土壤中筛选对PCNB具有降解效能的菌株(S1、S2、S3),研究其对PCNB的降解效能及影响因素.结果表明:适合各菌株生长的适宜温度为20～30%;适合S1和适合S2、S3菌株生长的pH范围分别为5.5～7.5和6.5～8.5.在此pH和温度范围内,各菌株对PCNB具有良好的降解性能.在最佳生长条件下,菌株达到最大生长量和最高降解率,在PCNB初始浓度为25.0 mg/L时,经过数天的培养,对PCNB降解率最高可达89.7%.与对照相比,土著微生物和接种微生物均能加速PCNB的降解.对泥浆相修复和纯培养基条件下的PCNB降解反应的回归方程进行差异显著性检验表明:在两种条件下,PCNB生物降解速率差异显著性达到α=0.01水平.     

反硝化技术对模拟养殖池塘修复的研究

浙江奉化市池塘的底泥经过反复培养、驯化,从中筛选、分离出反硝化细菌,在模拟实验条件下,研究其对不同浓度的硝酸盐氮和亚硝酸盐氮的去除情况,讨论反硝化菌种的生长情况。结果表明,在初始浓度为1、25、50mg·L^-1的硝酸盐氮和亚硝酸盐氮模拟池塘中,随着实验的进行,对污染物的去除效果逐渐提高。其中在1mg·L^-1的浓度组中,3d内硝酸盐氮和亚硝酸盐氮去除率就分别达到了95．8％和90．2％;在25mg·L^-1的浓度组中,第6d硝酸盐氮和亚硝酸盐的去除率分别为93．8％和87．8％;在50mg·L^-1的浓度组中,第6d硝酸盐氮和亚硝酸盐的去除率分别为89．7％和78．7％。此外,反硝化菌对硝酸盐氮的去除效果略优于亚硝酸盐氮,而且硝酸盐氮和亚硝酸盐氮的浓度越低,对其去除效果越好,达到稳定状态的时间越短。在模拟池塘中,菌种的生长情况与硝酸盐氮和亚硝酸盐氮的浓度呈负相关,即污染物的浓度越高反硝化菌的生长情况越差。对反硝化菌的生态影响因子研究表明,其反硝化最适宜的pH值为6～7,温度为25～35℃;而且在同一pH值和温度条件下,硝酸盐氮和亚硝酸盐氮浓度越低,反硝化菌对其去除效果越好。     

多级生物净化系统浮游植物的分析

利用不同生物的功能特性及其间的协同作用,由水生经济植物浮床、芦苇湿地、固定化微生物膜、底栖软体动物和滤食性鱼类组成了多级生物净化系统,除芦苇湿地、滤食性鱼类外,全部设置在多级生物净化水渠中。不仅充分利用了水面,节约了材料、能源,提高了净化效率,而且有效调控和净化了养殖池塘的水质,实现了养殖排放水的循环利用,提高了养殖效益。结果显示：养殖池塘排放水经过多级生物净化后,浮游植物的生物量控制在69．114-43．832mg／L,平均减少率为13．44％～0．47％,蓝藻平均减少率为58．41％-51．63％。     

除草剂阿特拉津的生态风险与植物修复研究进展

除草剂阿特拉津的生态毒性及其对人类健康、农业生产及生态环境的影响受到广泛关注。针对阿特拉津的使用现状和在环境中的残留及其危害,综述了阿特拉津的性质、分布和危害,介绍了阿特拉津污染土壤的植物修复及应用方面的研究进展。