Preliminary assessment of large‐scale co‐culture of sandfish (Holothuria scabra) with the Babylon snail (Babylonia areolata) in earthen ponds and in raceways

Holothuria scabra (sandfish) and Babylonia areolata were trialed in two large‐scale co‐culture experiments. Experiment 1 assessed co‐culture in 4 × 400 m² earthen ponds where Babylonia were cultured in a central pen at a density of 400 individuals/m² with sandfish occupying the remaining pond space at 1.1 individuals/m². Sandfish grew from 18.20 ± 6.67 g to 119.03 ± 17.74 g in 92 days and Babylonia (fed trash fish in both experiments) grew from 0.90 ± 0.38 g to 4.93 ± 1.44 g, but Babylonia growth was not increased in co‐culture compared to monoculture. Water and sediment quality varied between co‐culture and monoculture ponds. Neither showed clear improvement due to sandfish culture. Experiment 2 compared non‐segregated sandfish‐Babylonia co‐culture with Babylonia monoculture in 20 m² concrete raceways. Sandfish were cultured at 2 individuals/m² and Babylonia at 300 individuals/m². Sandfish grew up to 1.91 g.day^?1 with 100% survival. Babylonia weight gain was significantly greater in co‐culture raceways (3.35 ± 0.64 g over 61 days), which was double that of Babylonia in monoculture. Substrate total N was reduced by 20% in co‐culture compared with monoculture (p = .032). This provisional study of commercial scale sandfish‐Babylonia co‐culture demonstrates culture compatibility, providing a basis for further system development.     

Use of saline–sodic waters through phytoremediation of calcareous saline–sodic soils

Use of poor-quality groundwater has become inevitable for irrigation to compensate rapidly increasing water demands in many arid and semiarid regions. Salinity and sodicity are the principal soil and water quality concerns in such areas. Many saline–sodic and sodic soils have saline or saline–sodic subsurface drainage waters. Amelioration of these soils needs a source of calcium (Ca²⁺) that can replace the excess exchangeable sodium (Na⁺). Most of these soils, however, contain calcite (CaCO₃) of extremely low solubility. The native calcite does not supply adequate levels of Ca²⁺ for soil amelioration as do other chemical amendments. Phytoremediation may help ameliorate such soils through cultivation of certain crops tolerant to ambient soil salinity and sodicity. This amelioration strategy works through plant root action to help dissolve CaCO₃ to supply adequate Ca²⁺ without the application of an amendment. During a 3-year field experiment conducted under irrigated conditions, we evaluated phytoremediation against soil application of gypsum and farm manure, and water treatment with sulphuric acid on a calcareous saline–sodic soil (pH_s=8.0–8.4, EC_e=24–32 dS m⁻¹, SAR=57–78, CaCO₃=45–50 g kg⁻¹ for the top 0.15 m depth; Calcic Haplosalids). A saline–sodic water (EC=2.9–3.4 dS m⁻¹, SAR=12.0–19.4, RSC=4.6–10.0 mmol_c l⁻¹, SAR_adj=15.6–18.4) was used to irrigate the rice (Oryza sativa L.) and wheat (Triticum aestivum L.) crops grown in rotation. Active desalinisation and desodication processes were observed in all the treatments. After the final wheat crop, the 1.2 m soil profile EC_e was 7±0.5 dS m⁻¹ and SAR was 15±2 with non-significant treatment differences, indicating comparable soil amelioration effect of phytoremediation with other treatments. Better crop yields were obtained from the manure-treated plots, owing to its annual addition to the soil that possibly improved soil fertility. Phytoremediation needed minimum capital input because no initial investment was made to purchase the amendments.     

土壤生物修复机制探讨

指出了工业生产活动往往可能会对土壤环境造成污染,在土壤污染治理的各种技术中（如：换土、化学、生物、电修复、热修复等）,生物法因其成本低。效果显著而有较大优势。结合国内外的相关研究成果,对土壤生物修复的机制进行了探讨,并对土壤生物修复技术的应用进行了展望。     

Proton release by N2‐fixing plant roots: A possible contribution to phytoremediation of calcareous sodic soils

With a world‐wide occurrence on about 560 million hectares, sodic soils are characterized by the occurrence of excess sodium (Na⁺) to levels that can adversely affect crop growth and yield. Amelioration of such soils needs a source of calcium (Ca²⁺) to replace excess Na⁺ from the cation exchange sites. In addition, adequate levels of Ca²⁺ in ameliorated soils play a vital role in improving the structural and functional integrity of plant cell walls and membranes. As a low‐cost and environmentally feasible strategy, phytoremediation of sodic soils — a plant‐based amelioration — has gained increasing interest among scientists and farmers in recent years. Enhanced CO₂ partial pressure (P_CO2) in the root zone is considered as the principal mechanism contributing to phytoremediation of sodic soils. Aqueous CO₂ produces protons (H⁺) and bicarbonate (HCO₃^‐). In a subsequent reaction, H⁺ reacts with native soil calcite (CaCO₃) to provide Ca²⁺ for Na⁺ Ca²⁺ exchange at the cation exchange sites. Another source of H⁺ may occur in such soils if cropped with N₂‐fixing plant species because plants capable of fixing N₂ release H⁺ in the root zone. In a lysimeter experiment on a calcareous sodic soil (pH_s = 7.4, electrical conductivity of soil saturated paste extract (EC_e) = 3.1 dS m^‐1, sodium adsorption ratio (SAR) = 28.4, exchangeable sodium percentage (ESP) = 27.6, CaCO₃ = 50 g kg^‐1), we investigated the phytoremediation ability of alfalfa (Medicago sativa L.). There were two cropped treatments: Alfalfa relying on N₂ fixation and alfalfa receiving NH₄NO₃ as mineral N source, respectively. Other treatments were non‐cropped, including a control (without an amendment or crop), and soil application of gypsum or sulfuric acid. After two months of cropping, all lysimeters were leached by maintaining a water content at 130% waterholding capacity of the soil after every 24±1 h. The treatment efficiency for Na⁺ removal in drainage water was in the order: sulfuric acid > gypsum = N₂‐fixing alfalfa > NH4NO3‐fed alfalfa > control. Both the alfalfa treatments produced statistically similar root and shoot biomass. We attribute better Na+ removal by the N₂‐fixing alfalfa treatment to an additional source of H⁺ in the rhizosphere, which helped to dissolve additional CaCO₃ and soil sodicity amelioration.     

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.     

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.     

耐镉阿氏芽孢杆菌缓解水稻受镉胁迫的研究

为了探究阿氏芽孢杆菌T61缓解水稻受镉胁迫的效应,运用96孔板法、电感耦合等离子体质谱法、比色法测定菌株T61的镉耐受性、镉去除率和植物促生性;利用绿色荧光蛋白标记的T61,观察菌株在水稻上的定殖;利用蘸根法施加T61菌剂,观察其降低水稻籽粒镉积累的效应。结果表明：菌株T61对Cd²⁺的最大耐受浓度达到500 μmol·L^-1;在含镉液体培养基中培养24 h后,菌株T61对Cd²⁺的去除率超过50%。菌株T61可以合成植物促生性物质吲哚乙酸（6.2 μg·mL^-1）和铁载体（46.6 μmol·L^-1）,并具有溶磷能力（37.1 μg·mL^-1）。菌株T61可以在水稻根和茎上定殖。大田条件下,T61菌剂可以降低营养期水稻茎叶丙二醛含量和抗氧化酶活性,并使水稻728B和NX1B籽粒中的镉含量分别降低13.5%和11.2%。研究表明,阿氏芽孢杆菌T61是一株具有植物促生性的耐镉细菌,可以缓解某些水稻品种遭受的镉胁迫,在镉污染稻田的微生物修复方面具有一定的应用前景。     

养殖池塘污染底泥生物修复的室内比较实验

在实验室模拟生态条件下,运用投放复合微生物、微生物合酶菌液、添加营养促生剂、水底界面曝气等不同方法对养殖池塘污染底泥进行生物-生态修复;检测底泥及上覆水在不同生物修复技术作用下溶氧、氨氮、硝态氮、CODcr、TOC、底泥生物降解能力(G值)、异养细菌数量和反硫化细菌数量等指标的动态变化,据此评价不同修复技术的生态效应。结果表明,4种不同的生物生态方法均对污染底泥产生了一定的修复作用,其中以VB997底泥营养促生剂组的综合修复效果最为理想,在为期60 d的实验中,底表水CODcr消除率达70.00%,底泥生物降解能力(G值)从12.00 kg/(kg.h)提高至45.60 kg/(kg.h),底泥表面形成1.80~2.20 cm灰白色氧化层。曝气组溶氧充足,底层DO一直保持在7.00mg/L以上,底表水CODcr消除率达69.25%,G值从14.30 kg/(kg.h)升高到34.20 kg/(kg.h)。实验同时表明,几种生物制剂和营养促生剂的添加能导致上覆水硝态氮和氨态氮含量的升高,促进浮游藻类的阶段性滋生。[中国水产科学,2006,13(1):140-145]     

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.     

石油烃类污染物降解动力学和微生物群落多样性分析

为了探讨不同初始浓度石油污染土壤堆腐化修复机制,以石油降解菌剂和腐熟鸡粪为调理剂,研究了初始浓度分别为5 000（T1）、10 000（T2）和50 000 mg/kg（T3）的石油污染土壤堆腐化修复过程石油烃类污染降解动力学特征和微生物群落多样性。结果表明：堆腐化修复过程石油烃类污染物降解符合一级反应动力学,反应常数分别为0.012、0.094和0.050 d-1,半衰期分别为6.79、7.37和13.86 d。整个堆腐过程石油烃类污染物平均降解速率分别为112.08、230.05和887.93 mg/(kg·d)。3个处理的孔平均颜色变化率（average well color development）和碳源利用率（除芳香烃类化合物外）随堆腐进程的推进逐渐升高,在堆腐中、后期达到最大,T3处理显著高于T1、T2处理。多聚物类和糖类代谢群是堆腐体系中的优势菌群。主成分分析表明3个处理的微生物群落差异显著（除第9天外）,起分异作用的碳源主要是糖类和羧酸类。微生物群落的丰富度指数和均一度指数随堆腐进程的推进逐渐升高并在堆腐后期达到最大,与T1处理相比,T3处理分别高了0.21%和17.64%,差异达到显著水平（P<0.05）。微生物群落优势度指数在中期达到最大,T1处理分别比T2、T3处理高2.12%和9.44%,3个处理间差异不显著（P>0.05）。堆肥结束时3个处理的种子发芽指数（seed germination index, SGI）分别比堆腐初期提高了18.26%、20.42%和36.41%。该研究结果为黄土高原不同程度石油污染土壤堆腐化修复的应用提供参考依据和理论基础。