PERFORMANCE AND PERSISTENCE OF GREEN FLUORESCENT PROTEIN （gfp） MARKED AZOTOBACTER CHROOCOCCUM IN STERILIZED AND UNSTERILIZED WHEAT RHIZOSPHERIC SOIL

ThepracticaluseofAzotobacterchroococcumasbiofertilizer hasbeenincreasinglyidentifiedinrecentyears.Oftentheefficacy ofthesebacteriaisassessedonthebasisofcropresponseswithout knowingthesurvival,persistenceandcompetitiveabilityofthein troducedstrain.Inoculat…     

生物强化系统微生物分子诊断技术的应用及新发展

现代分子生物技术的飞速发展及其在环境研究领域的应用，为生物强化技术的研究和发展提供了新方法和新思路。本文从生物强化系统特异微生物检测及定量化技术、生物强化系统微生物群落结构组成及动态演替规律研究、生物强化作用机制的分子生物学解析、生物强化菌的基因工程构建、生物强化系统微生物的安全释放及控制技术几方面，对生物强化系统微生物分子诊断技术的应用和发展作了较为全面的综述。     

火山岩填料曝气生物滤池的SNAD工艺启动特性及功能菌丰度演替

为研究同步亚硝化、厌氧氨氧化耦合反硝化(SNAD)工艺在浸没式生物滤池反应器(SBAF)内的运行特性,同时接种亚硝化污泥和富集ANAMMOX的填料启动SNAD反应器.结果表明在60 mg·L~(-1)有机物浓度下,自养脱氮和反硝化实现较好的耦合,并在该浓度下稳定运行了67 d,其总氮去除率最高可达92.0%,COD去除率最高达82.9%,最高总氮去除负荷为2.3 kg·(m~3·d)~(-1).与全程自养脱氮(CANON)工艺相比,SNAD工艺的平均总氮去除率提高了12.6%.荧光定量PCR结果显示,系统启动后AOB菌的丰度有所增长,ANAMMOX菌的丰度增长了1个数量级,而NOB菌和反硝化菌的数量维持在较低水平(小于10~7 copies·g~(-1)),表明以火山岩为填料的浸没式生物滤池反应器有利于ANAMMOX和AOB的协同生长,可快速实现SAND工艺的启动.     

A bioaugmentation failure caused by phage infection and weak biofilm formation ability

Two biological aerated filters (BAF) were setup for ammonia removal treatment of the circulation water in a marine aquaculture. One of the BAFs was bioaugmented with a heterotrophic nitrifying bacterium, Lutimonas sp. H10, where the ammonia removal was not improved and the massive inoculation was even followed by a nitrification breakdown from day 9 to 18. The nitrification was remained stable in control BAF operated under the same conditions. Fluorescent in situ hybridization (FISH) with rRNA-targeted probes and cultivable method revealed that Lutimonas sp. H10 almost disappeared from the bioaugomented BAF within 3 d, and this was mainly due to the infection of a specific phage as revealed by flask experiment, plaque assay and transmission electron observation. Analyses of 16S rRNA gene libraries showed that bacterial groups from two reactors evolved di erently and an overgrowth of protozoa was observed in the bioaugmented BAF. Therefore, phage infection and poor biofilm forming ability of the inoculated strain are the main reasons for bioaugmentation failure. In addition, grazing by protozoa of the bacteria might be the reason for the nitrification breakdown in bioaugmented BAF during day 9–18.     

进水浓度或HRT变化时微好氧处理微生态机制研究

考察改变进水COD浓度（11 000、 15 000、 30 000 mg/Ｌ）或水力停留时间（HRT为42 h、 25 h）对模拟高浓度有机废水微好氧连续小试处理效果的影响,并采用荧光原位杂交-流式细胞术（FISH-FCM）、聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE) 等分子生物学技术及Biolog　FF微孔板法分析了不同处理阶段稳定期曝气柱污泥的微生态变化,以探讨进水COD浓度或HRT改变时微好氧处理效果变化的微生态机制.结果表明,在全部4个处理阶段,曝气柱污泥中酵母含量均保持在＞99.9％的水平.随着进水COD浓度的上升,污泥浓度由2.0 g/Ｌ上升到7.3 g/Ｌ,COD比去除速度由2.3 kg/（kg·d）下降到1.7 kg/（kg·d）,曝气柱污泥真菌群落结构多样性指数由2.05上升至2.19,代谢多样性指数由4.42上升至4.45;而随着HRT下降,污泥浓度从7.3 g/Ｌ下降至6.0 g/Ｌ,COD比去除速度从1.7 kg/（kg·d）上升至2.8 kg/（kg·d）,真菌群落结构多样性指数和代谢多样性指数则分别从2.19和4.45降至0.79和4.36.作为提高进水有机负荷的主要措施,提高进水COD浓度和缩短HRT对于废水处理效果和曝气柱微生态存在截然相反的影响.     

Do land utilization patterns a ect methanotrophic communities in a Chinese upland red soil?

Soil samples were collected from three plots under di erent land utilization patterns including degradation, farming, and restoration. The abundances of methanotrophs were quantified using real-time polymerase chain reaction (PCR) based on the pmoA and 16S rRNA genes, and the community fingerprint was analyzed using denaturing gradient gel electrophoresis (DGGE) aiming at pmoA gene. Significantly lower 16S rRNA and pmoA genes copies were found in the degradation treatment than in farming and restoration. Higher abundances of Type I than those of Type II methanotrophs were detected in all treatments. The treatment of farming was clearly separated from degradation and restoration according to the DGGE profile by cluster analysis. The lowest diversity indices were observed in the F (farming plot), suggesting that the community structure was strongly a ected by farming activities. There were significantly positive correlations between the copy numbers of pmoA also Type II-related 16S rRNA genes and soil available K content. Strong negative and positive correlations were found between Type I and soil pH, and available P content, respectively. We concluded that the vegetation cover or not, soil characteristics including pH and nutrients of P and K as a result of anthropogenic disturbance may be key factors a ecting methanotrophic communities in upland soil.     

CSTR系统发酵产氢细菌群落动态与产氢能力关系解析

将CSTR系统内pH由4.2一次性提高至6.0左右,启动发酵类型的转化,研究了转化过程系统内的产氢动态和细菌群落.结果表明,在有机负荷维持在（33±1）kg/(m³·d)的情况下,发酵类型10 d内未发生改变,产氢量8 d内未降低,15 d后系统内种群由乙醇型转化为丁酸型,进水碱度由250 mg/L增至2 450　mg/L.研究中利用荧光原位杂交技术（FISH）对反应系统内3类微生物群进行监测发现,在转化过程中Clostridium cluster XI数量增加,Clostridium cluster Ⅰ 和Ⅱ数量减少,而Enterobacteriaceae始终存在,变化不明显.种群的消长同反应系统产氢能力的高低存在密切关联,以Clostridium cluster Ⅰ和Ⅱ占优势的乙醇型发酵具有更佳的产氢能力,平均比产氢速率为23.6 mol/(kg·d).     

云南洱海沉积物中硫酸盐还原菌的时空分布特征

利用PCR扩增和荧光原位杂交方法分析了云南洱海沉积物中硫酸盐还原菌(Sulfate-reducing bacteria,SRB)类群和数量的时空分布特征。结果表明,秋季沉积物中检测出四个SRB类群(脱硫肠菌属、脱硫叶菌属、脱硫球菌属-脱硫线菌属-脱硫八叠菌属和脱硫弧菌属-脱硫微菌属),而春季沉积物中只检测出三个类群(脱硫肠菌属、脱硫叶菌属和脱硫球菌属-脱硫线菌属-脱硫八叠菌属)。相比春季,秋季沉积物检测出来的SRB类群具有更大的分布范围,同时,检测出的SRB类群主要分布在沉积物上部;从数量上来看,秋季沉积物SRB数量高于春季沉积物。上述结果指示秋季沉积物环境条件更适于SRB群落。此外,相比洱海秋季,洱海春季沉积环境溶解氧含量较高,且温度较低,这可能导致脱硫弧菌属-脱硫微菌属在洱海春季沉积物中处于不可育状态,由于其活性低而无法检测。     

型煤燃烧固硫中的温度效应

以氧化钙作为固硫剂的型煤中适当加入Ｆｅ,Ｓｉ添加剂组分,燃烧生成新的固硫体系后进行了２种不同温度条件下的进样实验。１．型煤样品分成６份,进行逐渐升温实验,分别加热到１１７０℃－１２２０℃,温度间隔１０℃,恒温１／４ｈ,２．型煤样品分成７份,进行先升温后进样实验,将加热炉分别升温至要求温度１０００℃－１２２０℃＋后进样,温度间隔５０℃,恒温１／２ｈ,实验表明：在Ｆｅ,Ｓｉ体系的型煤燃烧过程中,当进样     

绿色荧光蛋白标记在生物修复中的应用

通过对铜绿假单胞菌进行增强型绿色荧光蛋白(EGFP)标记,获得带有EGFP遗传标记的铜绿假单胞菌.该菌株绿色荧光标记性能稳定,通过转化子基因组DNAPCR和斑点杂交检测,证明外源EGFP基因存在于转化子染色体上.转化子30h内将溶液的表面张力由70mN/m降至35mN/m,产生的生物表面活性剂性能良好.