碳酸盐与磷酸盐对Biohydrogenbacterium R3sp.nov.产氢发酵效能的影响

采用NH4HCO3、NaHCO3、Na2CO3、K2HPO4、KH2HPO4、NaH2PO4和Na2HPO4作为碳酸盐和磷酸盐缓冲溶液源,研究其对Biohydrogenbacterium R3 sp.nov.产氢发酵效能的影响.研究发现,碳酸盐对Biohydrogenbacterium R3 sp.nov.反应体系中的pH值都具有较好的缓冲作用,而磷酸盐的添加对其产氢效能的促进作用较明显.反应器内添加了Na2HPO4与K2HPO4的Biohydrogenbacterium R3 sp.nov.的氢气产量与氢气含量达到最高值,分别为1978.56 mL、44.1％与2160.9 mL、45.8％,此时反应器内的pH值分别为3.28和3.41,细胞浓度分别为1.03 g·L-1和1.21 g·L-1,添加了Na2HPO4的Biohydrogenbacterium R3 sp.nov.的乙醇和乙酸产量分别为4841.49 mg·L-1和2064.94 mg·L-1;而添加了Na2CO3的氢气产量、氢气含量、细胞浓度与反应器内pH值分别为1064.42 mL、35.96％、1.23 g·L-1与3.81,此时乙醇、乙酸的产量分别为3862.21 mg·L-1和1930.86 mg·L-1.     

不同阴离子在生物质发酵产氢中的作用研究

为揭示不同阴离子在生物质发酵产氢中的作用规律,以混合菌群发酵产氢为研究对象,采用正交试验设计的方法考察了6种不同的阴离子及添加尿素对发酵产氢过程的影响.通过对不同阴离子浓度及相互关系对发酵产氢的影响的综合比较,获得以下结论:促进混合菌群发酵产氢的阴离子组成为:SO42-1.5 g·L-1,Cl-0 g·L-1,NO3-0 g·L-1,Mo7O246-0.1 g·L-1,C2O42- 1g·L-1,C6H5O73- 5g·L-1,(H2N)2CO 0 g·L-1;以PDB(马铃薯水解液)为基质发酵原料,H2最高含量和平均含量分别比对照组提高了21.4%和24.1%;平均产氢速率由对照组的36.79 mL·L-1·h-1提高到96.25 mL·L-1·h-1;氢气产率由对照组的0.76 mol·mol-1提高到1.17 mol·mol-1.研究同时证明,尿素对H2的产生具有显著的抑制作用,Mo7O246-则可促进混合微生物对原料的利用和转化;阴离子C2O42-和C6H5O73-的交互作用显著,说明两者同时使用可改善发酵产氢.从理论上对上述结果进行了分析探讨.     

利用巨藻发酵联产氢气与挥发性有机酸的研究

通过批式实验探讨了巨藻生物质厌氧发酵过程中巨藻的预处理方法、接种比、初始p H值等条件对累积产氢和挥发性脂肪酸(VFA)的影响,结果发现,巨藻可以用来发酵联产氢气与VFA;热碱预处理比较适于巨藻发酵产氢与产酸,在4 g·L-1的Na OH和100℃处理后,接种比为0.3,初始p H值为6时,单位挥发性固体(VS)最大累积产氢量可达到36.21 m L·g-1,比未经处理巨藻提高了77.82%;同时,巨藻在最佳的预处理条件下单位VS的总挥发性脂肪酸TVFA的产率为0.15 g·g-1,且以乙酸和丁酸为主.     

生物表面活性剂产生菌犁头霉菌(Absidia orchidis)的筛选及发酵条件优化

采用植物油为唯一碳源,设计选择培养基,从饭店下水道污泥中筛选出生物表面活性剂产生菌.结果分离到12株菌,其中一株能使发酵液的表面张力值从68 mN·m-1降低到34.5 mN·m-1,具有开发潜力,被选出作进一步的研究.该菌株经鉴定为犁头霉菌(Absidia orchidis).通过正交试验对犁头霉菌的培养条件进行优化,其优化培养条件为:植物油3.6 g·L-1,KH2PO412.1 g·L-1,Na2HPO45 g·L-1,(NH4)2SO4 1 g·L-1,NaNO32 g·L-1,酵母浸膏0.1 g·L-1,MgSO4·7H2O 0.15 g·L-1,NaCl 5 g·L-1,CaCl2 0.1 g·L-1,EDTA 1 g·L-1,KI 0.83μg·L-1,H3PO4 0.01μg·L-1,CoCl2·6H2O 0.048μg·L-1,MnSO4·H2O 0.312μg·L-1,Na2 MoO4·2H2O 0.048μg·L-1,ZnCl2 0.287μg·L-1,CuSO4·5H2O 0.125μg·L-1,初始pH值8,接种量6%.发酵70h时可获得生物表面活性剂的最大收获量,此时发酵液中生物表面活性剂的相对浓度达402倍.     

1株转座子插入突变菌株TB34的筛选及产氢分析

以分离自红树林污泥的厌氧发酵产氢细菌Pantoea agglomerans BH18为出发菌株,利用转座子Tn7构建突变体文库.通过卡那霉素抗性筛选与PCR扩增,鉴定转座子插入突变菌株.通过初筛和复筛,获得1株突变菌TB34,其产氢量较野生菌株明显提高.在初始pH为7.0和葡萄糖浓度10 g.L-1的海水培养条件下,产氢量(H2/葡萄糖)为(2.04±0.04)mol.mol-1,相比野生菌株产氢量提高43%.经过5次连续传代培养,突变菌株TB34表现出稳定的产氢特性.测定突变菌株TB34在不同碳源培养条件下的产氢量.结果表明,突变菌株TB34和野生菌株BH18都能利用蔗糖、葡萄糖和果糖发酵产氢.与野生菌株BH18不同,突变菌株TB34在以木糖为底物培养条件下仍能够发酵产氢,产氢量(H2/木糖)为(1.34±0.09)mol.mol-1,扩大了底物利用范围.     

一株高效产氢突变体RF-9的筛选与产氢特性

以从连续流搅拌槽式反应器(CSTR)内的活性污泥中分离出的厌氧产氢菌Ethanologenens sp.ZGX4为出发菌株,经过紫外线和亚硝酸复合诱变,从大量的突变体中筛选出一株稳定的高效产氢菌株RF-9.磷酸盐浓度在120mmol/L、温度37℃、初始pH6.0、葡萄糖浓度10g/L培养条件下,RF-9的单位体积产氢量和氢气产率为139.7mmol/L和2.52mol H2/mol葡萄糖,分别是对照的1.50倍和1.43倍.在发酵时间为15h、pH4.1、细胞干重0.722g/L时,RF-9获得其最大产氢速率345mLH₂/(h·L),是对照ZGX4的1.41倍.RF-9的主要液相末端产物为乙醇和乙酸,为典型的乙醇发酵细菌,与对照相似.     

一株耐酸产氢突变株Pantoea agglomerans的筛选与产氢特性

以红树林污泥中分离的厌氧发酵产氢细菌Pantoea agglomerans BH18为出发菌株,利用转座子Tn7随机插入菌株基因组DNA,通过卡那霉素筛选与PCR扩增验证,获得一批转座子插入突变菌株.起始pH4.0培养条件下,以产氢量为指标分离获得一株耐酸产氢突变菌株TB220.多次传代结果表明,突变菌株TB220具有稳定的产氢遗传特性.起始pH3.5~7.0范围内,突变菌株TB220最适产氢pH值为6.0,产氢量为(2.39±0.08)mol H2/mol葡萄糖.起始pH4.0和葡萄糖浓度10g/L的海水培养条件下,突变菌株TB220产氢量为(0.47 ± 0.02)mol H2/mol葡萄糖,比野生菌株高70%,表现出较强耐酸性.     

丁酸梭菌YM-83产氢影响因素的研究

丁酸梭菌是一株厌氧发酵产氢细菌,可以参与高浓度有机废水的厌氧发酵过程,在降解高浓度有机废水的同时制取氢能源。为了确定L-cys和金属离子对丁酸梭菌YM-83产氢能力的影响,文章通过间歇实验对其进行产氢培养优化,确定其最适产氢的L-cys和金属离子浓度。研究结果表明,丁酸梭菌YM-83的最适生长和产氢的L-cys浓度均为1.0 g/L,最适生长和产氢的Mg~(2+)浓度均为0.15g/L,最适生长和产氢的铁元素类型均为Fe~(2+),其最适浓度均为0.2 g/L。在此条件下,获得的最大比产氢率为2.15 mol H2/mol glucose,单位体积产氢量为5 360 mL/L culture,细胞干重为1.04 g/L。丁酸梭菌YM-83是一株产氢能力良好的厌氧发酵产氢细菌,可以实现厌氧发酵的高效产氢。     

絮凝天然碱碱泥的絮凝剂产生菌的筛选及最佳产絮条件研究

从生活污水中分离得到一株在强碱性条件下能絮凝天然碱碱泥的高效絮凝剂产生菌EWY1,并通过稳定性试验证明了其具有较好的稳定性,经鉴定为肠球菌属(Enterococcus)中的盲肠肠球菌(E.cecorum).通过单因素试验和正交试验确定了该菌产絮凝剂的最佳培养基为:麦芽糖15 g·L-1,KNO3 1.2 g·L-1,KH2PO4 0.5 g·L-1,MgSO4·7H2O 0.4 g·L-1,NaCl0.6 g·L-1,FeSO4·7H2O 0.008 g·L-1;该菌产絮凝剂的最佳培养条件为pH 7.0、温度30℃、摇床转速170 r·min-1.絮凝实验结果表明,用该菌产生的絮凝剂处理天然碱碱泥效果较好,絮凝率达到了89%.     

混合碳源制备生物絮凝剂的絮凝效能及产量预测模型

为实现混合底物的高效定向转化,以产絮菌根癌农杆菌(Agrobactrium tumefaciens)F2为研究对象,考察不同单一碳源及不同初始浓度对菌体生长、絮凝效能及絮凝剂产量的变化规律,采用BP算法构建絮凝效能及产量预测神经网络.产絮菌F2利用葡萄糖时的絮凝效能和产量分别为88.98%和2.20 g·L-1,过低的初始浓度将影响产量,不低于7.5 g·L-1为佳.以D-(+)-葡萄糖、D-半乳糖和D-甘露糖3种单糖为混合碳源,构建网络结构为3-5-2的产絮效能及絮凝剂产量预测模型,对两个输出层的预测误差范围均在4%以内,预测葡萄糖、半乳糖、甘露糖浓度的最优解为6.59 g·L-1、1.32 g·L-1、3.57 g·L-1,经验证混合碳源发酵产絮可使絮凝效能和产量比单一葡萄糖发酵时分别提高6.87%和26.82%,本文为产絮菌F2利用含糖有机质废液发酵产絮凝剂提供数据参考.     

Effects of two sludge application on fractionation and phytotoxicity of zinc and copper in the soil

The potential harm of heavy metals is a primary concern in application of sludge to the agricultural land. A pot experiment was conducted to evaluate the effect of two sludges on fractionation of Zn and Cu in soil and their phytotoxicity to pakchoi. The loamy soil was mixed with 0%, 20%, 40%, 60% and 80% (by weight) of digested sewage sludge (SS) and composted sludge (SC). The additions of both sludges caused a significant raise in all fractions, resulting in that exchangeable (EXCH) and organic bound (OM) became predominance of Zn and organic bound Cu occupied the largest portion. There was more available amount of Zn and Cu in SS treatments than SC treatments. During the pot experiment, the concentration of Zn in EXCH, carbonate (CAR) and OM and Cu in EXCH and OM fractions decreased in all treatments, so their bioavailability reduced. Germination rate and plant biomass decreased when the addition rate was high and the best yield appeared in 20% mixtures at the harvest of pakchoi. The two sludges increased tissue contents of Zn and Cu especially in the SS treatments. Zn in pakchoi was not only in relationship to ΔEXCH and ΔCAR forms but also in ΔOM forms in the sludge-soil mixtures. Tissue content of Cu in pakchoi grown on SC-soils could not be predicted by ΔEXCH. These correlation rates between Zn and Cu accumulation in pakchoi and variation of different fractions increased with time, which might indicate that sludges represented stronger impacts on the plant in long-term land application.     

Intraspecific differences in effects of co-contamination of cadmium and

A hydroponic experiment was carried out to study intraspecific differences in the effects of different concentrations of cadmium (Cd)(0-10 mg/L) and arsenate (As(V)) (0-8 mg/L) on the growth parameters and accumulation of Cd and As in six wheat varieties Jing-9428, Duokang-1, Jingdong-11, Jing-411, Jingdong-8 and Zhongmai-8. The endpoints of wheat seedlings, including seed germination,biomass, root length and shoot height, decreased with increasing the Cd and As concentrations. Significant differences in seed germination, biomass, root length, shoot height and the accumulation of Cd and As were observed between the treatments and among the varieties (p ＜ 0.05). The lethal dosage 50% were about 20, 80, 60, 60, 80 and 20 mg As/L for Jing-9428, Duokang-1, Jingdong-11,Jing-411, Jingdong-8 and Zhongmai-8, respectively, and the corresponding values for Cd were about 30, 80, 20, 40, 60 and 10 mg Cd/L, respectively. Among the six varieties, Duokang-1 was found to be the most resistant to Cd and As toxicity, and Zhongmai-8 was the most sensitive to Cd and As co-contamination. The resistance of the six varieties was found dependant on the seedling uptake of Cd and As. Duokang-1 was the most suitable for cultivation in Cd and As co-contaminated soils.     

Toxic effects of acetochlor, methamidophos and their combination on nifH gene in soil

Toxic effects of two agrochemicals on nifH gene in agricultural black soil were investigated using denaturing gradient gel electrophoresis （DGGE） and sequencing approaches in a microcosm experiment. Changes of soil nifH gene diversity and composition were examined following the application of acetochlor, methamidophos and their combination. Acetochlor reduced the nifH gene diversity （both in gene richness and diversity index values） and caused changes in the nifH gene composition. The effects of acetochlor on nifH gene were strengthened as the concentration of acetochlor increased. Cluster analysis of DGGE banding patterns showed that nifH gene composition which had been affected by low concentration of acetochlor （50 mg/kg） recovered firstly. Methamidophos reduced nifH gene richness that except at 4 weeks. The medium concentration of methamidophos （150 mg/kg） caused the most apparent changes in nifH gene diversity at the first week while the high concentration of methamidophos （250 mg/kg） produced prominent effects on nifH gene diversity in the following weeks. Cluster analysis showed that minimal changes of nifH gene composition were found at 1 week and maximal changes at 4 weeks. Toxic effects of acetochlor and methamidophos combination on nifH gene were also apparent. Different nifH genes （bands） responded differently to the impact of agrochemicals： four individual bands were eliminated by the application of the agrochemicals, five bands became predominant by the stimulation of the agrochemicals, and four bands showed strong resistance to the influence of the agrochemicals. Fifteen prominent bands were partially sequenced, yielding 15 different nifH sequences, which were used for phylogenetic reconstructions. All sequences were affiliated with the alpha- and beta-proteobacteria, showing higher similarity to eight different diazotrophic genera.     

Arsenic uptake by arbuscular mycorrhizal maize （Zea mays L.） grown in an arsenic-contaminated soil with added phosphorus

The effects of arbuscular mycorrhizal (AM) fungus (Glomus mosseae) and phosphorus (P) addition (100 mg/kg soil) on arsenic (As) uptake by maize plants (Zea mays L.) from an As-contaminated soil were examined in a glasshouse experiment.Non-mycorrhizal and zero-P addition controls were included.Plant biomass and concentrations and uptake of As,P,and other nutrients,AM colonization,root lengths,and hyphal length densities were determined.The results indicated that addition of P significantly inhibited root colonization and development of extraradical mycelium.Root length and dry weight both increased markedly with mycorrhizal colonization under the zero-P treatments,but shoot and root biomass of AM plants was depressed by P application.AM fungal inoculation decreased shoot As concentrations when no P was added,and shoot and root As concentrations of AM plants increased 2.6 and 1.4 times with P addition,respectively.Shoot and root uptake of P,Mn,Cu,and Zn increased,but shoot Fe uptake decreased by 44.6%,with inoculation, when P was added.P addition reduced shoot P,Fe,Mn,Cu,and Zn uptake of AM plants,but increased root Fe and Mn uptake of the nonmycorrhizal ones.AM colonization therefore appeared to enhance plant tolerance to As in low P soil,and have some potential for the phytostabilization of As-contaminated soil,however,P application may introduce additional environmental risk by increasing soil As mobility.     

Degradation of metabolites accumulated of benzo[a]pyrene by coupling Penicillium Chrysogenum with KMnO4

Several main metabolites of benzo[a]pyrene （BaP） formed by Penicillium chrysogenum, Benzo[a]pyrene-1,6-quinone （BP 1,6- quinone）, trans-7,8-dihydroxy-7,8-dihydrobenzo[a]pyrene （BP 7,8-diol）, 3-hydroxybenzo[a]pyrene （3-OHBP）, were identified by high-performance liquid chromatography （HPLC）. The three metabolites were liable to be accumulated and were hardly further metabolized because of their toxicity to microorganisms. However, their further degradation was essential for the complete degradation of BaP. To enhance their degradation, two methods, degradation by coupling Penicillium chrysogenum with KMnO4 and degradation only by Penicillium chrysogenum, were compared; Meanwhile, the parameters of degradation in the superior method were optimized. The results showed that （1） the method of coupling Penicillium chrysogenum with KMnO4 was better and was the first method to be used in the degradation of BaP and its metabolites; （2） the metabolite, BP 1,6-quinone was the most liable to be accumulated in pure cultures; （3） the effect of degradation was the best when the concentration of KMnO4 in the cultures was 0.01% （w/v）, concentration of the three compounds was 5 mg/L and pH was 6.2. Based on the experimental results, a novel concept with regard to the bioremediation of BaP-contaminated environment was discussed, considering the influence on environmental toxicity of the accumulated metabolites.     

Impact of atrazine and nitrogen fertilizers on the sorption of chlorotoluron in soil and model sorbents

Sorption of chlorotoluron in ammonium sulfate, urea and atrazine multi-solutes system was investigated by batch experiments. The results showed application of nitrogen fertilizers to the soil could affect the behavior of chlorotoluron. At the same concentration of N, sorption of chlorotoluron decreased as the concentration of atrazine increased on the day 0 and 6 in soil, respectively. The sorption of chlorotoluron increased from 0 to 6 d when soils were preincubated with deionized water, ammonium sulfate and urea solution for 6 d. That indicated incubation time was one of the most important factors for the sorption of chlorotoluron in nitrogen fertilizers treatments. The individual sorption isotherms of chlorotoluron in rubbery polymer and silica were strictly linear in single solute system, but there were competition sorption between pesticides or between pesticides and nitrogen fertilizers. That indicated the sorption taken place by concurrent solid-phase dissolution mechanism and sorption on the interface of water-organic matter or water-mineral matter.     

Distribution and ecological effect of mercury in Laogang landfill, Shanghai, China

Laogang landfill near Shanghai is the largest landfill in China, and receives about 10000 t of daily garbage per day, Samples of topsoil and plants were analyzed to evaluate mercury pollution from the landfill. For topsoil samples, there were significant correlations among total mercury （HgT）, combinative mercury （Hgc） and gaseous mercury （HgG）, and content of total organic carbon （TOC）, but, no significantly relationship was found between Hg content and filling time. Hg content changes in vertical profiles with time showed that the average Hgv of profiles 1992, 1996, and 2000 was similar, but their average HgG was quite different. HgT was significantly correlated with Hgc in profile 1992 and 2000, and Hgv was significantly correlated with Hg6 in profile 1996. HgG/Hgv ratio in profile samples decreased in the order of （HgG,/HgT）1992〉（HgG/HgT）1996〉〉（HgG/HgT）2000. A simple outline of Hg release in landfill could be drawn： with increasing of filling time, degradation undergoes different biodegradation, accordingly, gaseous mercury goes through small, more, and small proportion to total mercury. Distribution of Hg in plants was inhomogeneous, following the order of leaf〉root〉stem. The highest value of leaf may be associated with higher atmospheric Hg from landfill. Ligneous plants （e.g. Phyllostachys glanca, Prunus salicina and Ligustrum lucidum） are capable of enriching more Hg than herbaceous plants.     

EDTA-enhanced phytoremediation of lead contaminated soil by Bidens maximowicziana

Phytoremediation is a potential cleanup technology for the removal of heavy metals from contaminated soils.Bidens maximowicziana is a new Pb hyperaccumulator,which not only has remarkable tolerance to Pb but also extraordinary accumulation capacity for Pb.The maximum Pb concentration was 1509.3 mg/kg in roots and 2164.7 mg/kg in overground tissues.The Pb distribution order in the B. maximowicziana was:leaf>stem>root.The effect of amendments on phytoremediation was also studied.The mobility of soil Pb and the Pb concentrations in plants were both increased by EDTA application.Compared with CK(control check),EDTA application promoted translocation of Pb to overground parts of the plant.The Pb concentrations in overground parts of plants was increased from 24.23-680.56 mg/kg to 29.07-1905.57 mg/kg.This research demonstrated that B.maximowicziana appeared to be suitable for phytoremediation of Pb contaminated soil,especially,combination with EDTA.     

Potential of plant polyphenol oxidases in the decolorization and removal of textile and non-textile dyes

In this study an effort has been made to use plant polyphenol oxidases; potato （Solanum tuberosum） and brinjal （Solanum melongena）, for the treatment of various important dyes used in textile and other industries. The ammonium sulphate fractionated enzyme preparations were used to treat a number of dyes under various experimental conditions. Majority of the treated dyes were maximally decolorized at pH 3.0. Some of the dyes were quickly decolorized whereas others were marginally decolorized. The initial first hour was sufficient for the maximum decolorization of dyes. The rate of decolorization was quite slow on long treatment of dyes. Enhancement in the dye decolorization was noticed on increasing the concentration of enzymes. The complex mixtures of dyes were treated with both preparations of polyphenol oxidases in the buffers of varying pH values. Potato polyphenol oxidase was significantly more effective in decolorizing the dyes to higher extent as compared to the enzyme obtained from brinjal polyphenol oxidase. Decolorization of dyes and their mixtures, followed by the formation of an insoluble precipitate, which could be easily removed simply by centrifugation.     

Decomposition of alachlor by ozonation and its mechanism

Decomposition and corresponding mechanism of alachlor, an endocrine disruptor in water by ozonation were investigated. Results showed that alachlor could not be completely mineralized by ozone alone. Many intermediates and final products were formed during the process, including aromatic compounds, aliphatic carboxylic acids, and inorganic ions. In evoluting these products, some of them with weak polarity were qualitatively identified by GC-MS. The information of inorganic ions suggested that the dechlorination was the first and the fastest step in the ozonation of alachlor.