酸性矿山废水区域沉积物中嗜酸菌多样性研究

采集了酸性矿山废水区域酸性沉积物样品,运用16S rDNA克隆文库技术和变性梯度凝胶电泳(DGGE)技术,研究了沉积物中的嗜酸菌类群以及它们的群落结构.结果表明,沉积物中的嗜酸菌类型较新,大多数16S rDNA序列与已知序列的相似性均在97%以下.这些嗜酸菌主要为Acidobacteria,β/-γProteobacteria,δ-Proteobacteria,Nitrospira,Candidate Division TM7,low G+C Gram-positive类微生物.在这一酸性生态系统中,δ-Proteobacteria类细菌占据主导地位.对酸性沉积物中嗜酸菌多样性的研究有利于发现高效处理酸性矿山废水的方法.     

A/O工艺膜生物反应器处理生活污水的脱氮特性及硝化菌群的分子检测

采用A/O工艺膜生物反应器(MBR),以生活污水为处理对象,考察了系统的脱氮特性,并采用聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)与荧光原位杂交(FISH)技术对系统中硝化菌群进行了分子检测.结果表明,A/O工艺MBR处理生活污水,TN去除率在85%左右,NH3-N去除率在95%以上;DGGE图谱显示,随着系统运行时间的延长,硝化菌群数量逐渐增加,并且不同菌种的丰度也发生了变化;FISH检测显示,系统中硝化菌的优势菌种为氨氧化菌和亚硝酸氧化菌.应用Motic Fluo 1.0软件对FISH结果进行分析,结果显示,系统运行初期到末期,氨氧化菌占硝化菌的比例一直保持在25%左右;亚硝酸氧化菌占硝化菌的比例由系统运行初期的35%逐渐增加到系统运行末期的55%左右.     

活性污泥总DNA提取方法的比较

从处理某制药废水的MBR反应器中采集活性污泥,评价不同DNA提取方法对其总DNA提取效率的影响。DNA提取分细胞裂解和DNA纯化2步,对细胞裂解比较了珠磨匀浆法、反复冻融法、十二烷基磺酸钠（sodium dodecyl sulfate,SDS）裂解法等7种方法;对DNA纯化比较了酚/氯仿纯化法和胶回收纯化法。结果表明,SDS裂解法、酚氯仿纯化法最优。通过条件优化实验,确定SDS裂解酚/氯仿纯化法在污泥量1.1 g,10 000 r/min离心5 min的操作条件下,获得的DNA产量（10 774 μg/g泥重）和纯度（OD₂₆₀∶OD₂₈₀=1.84）等综合指标最好。     

Characterization of bacterial diversity in an atrazine degrading enrichment culture and degradation of atrazine,cyanuric acid and biuret in industrial wastewater

An enrichment culture was used to study atrazine degradation in mineral salt medium (MSM) (T1), MSM+soil extract (1:1, v/v) (T2) and soil extract (T3). Results suggested that enrichment culture required soil extract to degrade atrazine, as after second sequential transfer only partial atrazine degradation was observed in T1 treatment while atrazine was completely degraded in T2 and T3 treatments even after fourth transfer. Culture independent polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) technique confirmed selective enrichment of genus Bacillus along with Pseudomonas and Burkholderia. Degradation of atrazine/metabolites in the industrial wastewater was studied at different initial concentrations of the contaminants [wastewater-water (v/v) ratio: T1, 1:9; T2, 2:8; T3, 3:7; T4, 5:5 and T5, undiluted effluent]. The initial concentrations of atrazine, cyanuric acid and biuret ranged between 5.32 and 53.92 µg mL^?1, 265.6 and 1805.2 µg mL^?1 and 1.85 and 16.12 µg mL^?1, respectively. The enrichment culture was able to completely degrade atrazine, cyanuric acid and biuret up to T4 treatment, while no appreciable degradation of contaminants was observed in the undiluted effluent (T5). Inability of enrichment culture to degrade atrazine/metabolites might be due to high concentrations of cyanuric acid. Therefore, a separate study on cyanuric acid degradation suggested: (i) no appreciable cyanuric acid degradation with accumulation of an unidentified metabolite in the medium where cyanuric acid was supplemented as the sole source of carbon and nitrogen; (ii) partial cyanuric acid degradation with accumulation of unidentified metabolite in the medium containing additional nitrogen source; and (iii) complete cyanuric acid degradation in the medium supplemented with an additional carbon source. This unidentified metabolite observed during cyanuric acid degradation and also detected in the enrichment culture inoculated wastewater samples, however, was degraded up to T4 treatments and was persistent in the T5 treatment. Probably, accumulation of this metabolite inhibited atrazine/cyanuric acid degradation by the enrichment culture in undiluted wastewater.     

Effects of fungicides triadimefon and propiconazole on soil bacterial communities

The impact of fungicides triadimefon and propiconazole on soil bacterial populations from a strawberry field was investigated. Two fungicides were applied to the soil at concentrations of 10 mg/kg or 100 mg/kg with soil water contents 20.2% (fresh soil water content) or 26.0% (field capacity). Changes in bacterial communities were assessed using DNA extraction, polymerase chain reaction (PCR) amplification of the 16S rDNA and denaturing gradient gel electrophoresis (DGGE). High performance liquid chromatography (HPLC) was utilized to detect the residue of fungicides in soils. The results showed that propiconazole was more persistent than triadimefon in soils, and the two soil water contents did not cause significant differences in dissipation rates between the two fungicides. A high concentration of propiconazole could inhibit the existence of soil microbes while one of triadimefon might induce the microbial population in the first stage. From unweighted pair-group method using arithmetic averages (UPGMA) dendrograms, the effect of triadimefon and propiconazole at the two applied concentrations on a soil bacterial community could be long term. After triadimefon was applied for 60 days and propiconazole for 75 days, the compositions of microbial communities were not recovered. From the viewpoint of environmental protection, it was of significant importance to pay more attention not only to the residues of pesticide but also to the change in soil microbial communities.     

Effects of cultivation conditions on the diversity of microbes involved in the conversion of rice straw to fodder

To confirm the optimum cultivation conditions for analyzing lactic acid bacterial communities and to provide the cultivation foundation for lactic acid bacterial communities that were used to convert straw into fodder, fermented rice straw was inoculated into 13 different broths. After 48 h of cultivation, pH values, volatile products, and microbial diversity were analyzed. Except for LAB broth, the pH values of the other broths could decrease to approximately 4.5. GC/MS analysis showed that lactic acid in Tomato MRS broth, MRS broth, LAB broth, and Tomato juice broth was higher than that in the other broths. DNA concentration analysis showed that the counts of microbes in Tomato MRS broth were 2.5 times of those in other broths and that tomato juice favored the reproduction of the microbes. Denaturing gradient gel electrophoresis （DGGE） analysis showed that the number of lactic acid bacterial species in HYA broth, Tomato juice broth, and Tomato MRS broth were higher than those in the other broths.     

Compost: Its role,mechanism and impact on reducing soil-borne plant diseases

Soil-borne plant pathogens are responsible for causing many crop plant diseases, resulting in significant economic losses. Compost application to agricultural fields is an excellent natural approach, which can be taken to fight against plant pathogens. The application of organic waste products is also an environmentally friendly alternative to chemical use, which unfortunately is the most common approach in agriculture today. This review analyses pioneering and recent compost research, and also the mechanisms and mode of action of compost microbial communities for reducing the activity of plant pathogens in agricultural crops. In addition, an approach for improving the quality of composts through the microbial communities already present in the compost is presented. Future agricultural practices will almost definitely require integrated research strategies to help combat plant diseases.     

Vertical diversity of sediment bacterial communities in two different trophic states of the eutrophic Lake Taihu, China

Vertical diversity of sediment bacterial communities in 2 different trophic states (macrophyte-dominated and algae-dominated) of the large shallow eutrophic Lake Taihu, China, were investigated using denaturing gradient gel electrophoresis (DGGE) and 16S rRNA sequence analysis. Clustering analysis of DGGE profiles showed that different clusters were recognized in different depths of sediment cores in the 2 lake trophic states. Analyses of the bacterial diversity, as estimated by the Shannon index (H'), showed that different sediment layers of the macrophyte-dominated state had higher diversity than the algae-dominated state. In addition, bacterial diversity of the sediment in the macrophyte-dominated state changed abruptly throughout the layers, but bacterial diversity of the algae-dominated state decreased gradually with sediment depth. Phylogenetic analysis showed that Proteobacteria was the most abundant phylum in the middle sediment of the 2 lake trophic states. In the macrophyte-dominated state, clone sequences related to Betaproteobacteria (50.0%) were the most abundant, followed by Epsilonproteobacteria (21.1%), Acidobacteria (7.9%), Deltaproteobacteria (7.9%), Chloroflexi (7.9%), and Bacteroidetes (5.3%); whereas in the algae-dominated state, sequences affiliated with Betaproteobacteria (84.4%) were predominant, followed by Deltaproteobacteria (12.5%) and Acidobacteria (3.1%). Canonical correspondence analysis showed that organic matter and pH play key roles in driving the vertical changes of bacterial community composition.     

利用DGGE-菌落原位杂交法分离土壤中精喹禾灵降解菌

从自然环境中分离到的可降解农药的土著微生物,因其对环境的友好性及原位修复的可行性,受到了高度关注.为从土壤中筛选精喹禾灵降解菌株,首先利用PCR-DGGE技术分析了除草剂精喹禾灵胁迫下土壤细菌群落结构及多样性的变化.结果表明,添加精喹禾灵后,土壤细菌群落结构发生了明显的改变.精喹禾灵使细菌多样性呈现出增加-减少-增加的变化趋势,其中第9 d变化最大,后期趋于稳定.根据DGGE图谱条带的测序结果推断,Pseudomonas、Massilia、Burkholderia等属中的细菌对精喹禾灵具有耐受性或降解潜力,这些微生物类群可作为减少农药残留的土著微生物资源进行分离筛选.根据条带的测序结果,合成了地高辛(Digoxigenin)标记的探针,并进行了菌落原位杂交,筛选到了3株具有降解潜力的菌株,其中L1可以利用精喹禾灵作为唯一碳源生长,经16S rRNA基因鉴定该菌株属于假单胞菌属(Pseudomonas sp.).利用高效液相色谱法测定了菌株L1在无机盐培养基中降解精喹禾灵的效果.结果表明,培养7 d后,精喹禾灵的含量减少了近50%,且随着精喹禾灵含量的降低,L1菌体数量增加,证实了菌株L1具有降解精喹禾灵的能力.这一结果为今后研究菌株L1降解精喹禾灵的机制、功能基因等奠定了基础.     

生物滴滤池对BTEX的去除及相应细菌群落分析

以预先驯化的菌群和活性污泥作为起始接种物用于生物滴滤池(BTF)中,研究评估了BTF去除苯、甲苯、乙苯和二甲苯混合气体(BTEX)的性能,并利用变性梯度凝胶电泳(DGGE)技术分析了微生物群落结构的变化.结果表明,BTF能在短时间内得到驯化,填料附着的生物量从第10 d的5.7 mg.g-1迅速增加至第30 d的112 mg.g-1.BTF能同时有效去除混合BTEX中的各组分,在进气负荷和停留时间分别为269.7 g.(m3.h)-1和39 s时,可获得的最大去除能力为216.6 g.(m3.h)-1.DGGE图谱表明,BTF中优势微生物种群主要来源于富集菌群,微生物群落结构随着运行时间发生变化,但在BTF上下空间分布较为均匀.