Seasonal variation of microbial populations and biomass in Tatachia grassland soils of Taiwan

To investigate the seasonal variations of microbial ecology in grassland of Tatachia forest, soil properties, microbial populations, microbial biomass, and 16S rDNA clone library analysis were determined. The soil had temperatures 6.6–18.4°C, pH 3.6–5.1, total organic carbon 1.11–10.68%, total nitrogen 0.18–0.78%, and C/N ratios 3.46–20.55. Each gram of dry soil contained bacteria, actinomycetes, fungi, cellulolytic, phosphate-solubilizing microbes, and nitrogen-fixing microbes 4.54 × 10⁴ to 3.79 × 10⁷, 3.43 × 10² to 2.17 × 10⁵, 5.74 × 10³ to 3.76 × 10⁶, 1.97 × 10³ to 1.34 × 10⁶, 8.49 × 10² to 5.59 × 10⁵, and 3.86 × 10² to 3.75 × 10⁵ CFU, respectively. Each gram of soil contained 117–2,482 μg of microbial biomass carbon, 23–216 μg of microbial biomass nitrogen and 9–29 μg of DNA. The microbial populations, microbial biomass, and DNA decreased stepwise with the depth of soil, and they had low values in winter seasons. The microbial populations, microbial biomass carbon, microbial biomass nitrogen, and DNA at the BW2 horizon were 8.42–17.84, 19.26–64.40, 16.84–61.11, and 31.03–46.26% of those at the O horizon, respectively. When analyzing 16S rDNA library, members of Proteobacteria, Acidobacteria, Actinobacteria, Bacteroidetes, Chloroflexi, Firmicutes, candidate division TM1, candidate division TM7, Gammatimonadetes, and Verrucomicrobia were identified. Members of Proteobacteria (44.4%) and Acidobacteria (33.3%) dominated the clone libraries. Within the phylum Proteobacteria, α-, β-, and γ-Proteobacteria were most numerous, followed by δ-Proteobacteria.     

湖南衡阳地区乙型肝炎患者HBV基因分型与血清HBVDNA水平研究

分析了湖南衡阳地区乙型肝炎病毒基因型分布特征,并探讨HBV各基因型与血清HBVDNA水平之间的关系．采用荧光定量PCR结合TaqmanMGB探针技术,对湖南衡阳地区164份乙型肝炎患者血清中的HBVDNA进行基因分型和定量检测．结果164例血清中,C型110A．（67．1％）,B、C型37人（22．3％）,B型8人（5．1％）,D型1人（0．6％）,A型1人（0．6％）．C型与B、C混合型在HBVDNA水平上存在显著差异（t=2．433,P〈0．05）,B、C混合型的HBVDNA水平明显高于C型．由此得出湖南衡阳地区HBV基因型以C型为主、B、C混合型次之,B型较少,D型、A型极少;B、C混合型患者的HBVDNA数量水平显著高于C型．表1．参11．     

Assessment of groundwater contamination caused by uncontrolled dumping in old gravel quarries in the Besòs aquifers (Barcelona,Spain)

The contamination of groundwater in the aquifer of the La Llagosta basin (Besòs river basin) due to waste disposal in quarries formerly used for the extraction of dry raw materials has led to the cessation of groundwater extraction for public water supply. The mobilization of pollutants was largely caused by fluctuations in piezometric levels, which led to the washing of buried waste. The hydrogeochemical processes associated with uncontrolled waste disposal in these landfilled areas of the La Llagosta basin aquifer were studied along a flow path that crosses the contaminated area. The PHREEQC code was used to establish the reactions associated with the different mineral phases through inverse modeling. This transport code, ionic exchange phenomena, surface reactions and balance (mineral phase) reactions were used to simulate the dilution phenomenon associated with the pollution after the potential removal of the sources of contamination. One-dimensional advective–dispersive modeling indicates a substantial reduction in Ca, Mg, Na and SO₄²⁻ within one year and stabilization within four years.     

温带亚高山针叶林土壤碳循环微生物群落的时空动态

选取参与碳固定的二磷酸羧化/加氧酶基因（cbbM）、有机碳降解的淀粉酶基因（amylase）和纤维素酶基因（cellulase）作为分子标记,用实时定量PCR方法对温带亚高山华北落叶松（Larix gmelinii var.principis-rupprechtii）林、白杄（Picea meyeri）林、青杄（P.wilsonii）林和油松（Pinus tabulaeformis）林土壤碳循环功能微生物类群丰度的时空动态开展研究.结果显示,总碳（TC）、总氮（TN）、总硫（TS）、有机质（OM）和有机碳（TOC）、pH值、铵态氮（NH₄⁺-N）、硝态氮（NO₃^--N）、过氧化氢酶、蔗糖酶和脲酶活性在4种森林土壤中都有不同程度的差异,且有显著的季节变化特征.高海拔华北落叶松林土壤TC、TN、TS、C/N、OM和TOC含量最高,而pH值最低.土壤TC、TN、亚硝态氮（NO₂^--N）含量、蔗糖酶和脲酶活性,与碳循环微生物类群的丰度呈极显著相关.土壤NO₃^--N含量与有机碳分解和固碳微生物类群的相对丰度显著相关;土壤C/N、NO₂^--N、pH值、OM、TOC、过氧化氢酶及脲酶活性,与降解易分解碳（labile C）和难分解碳（recalcitrant C）的微生物类群的相对丰度呈极显著相关.植被类型和季节变化共同影响土壤碳循环微生物类群的丰度,而季节变化是主导因素.植被和土壤环境因子通过调控微生物群落碳代谢功能类群的结构,影响森林土壤碳源-汇的平衡.     

慢生花生根瘤菌的遗传多样性研究

用ＲＥＰ１Ｒ１和ＲＥＰ２１ＤＮＡ为引物,对２２株四川土著慢生花生根瘤菌染色体ＤＮＡ的基因外回文重复序列（ＲｅｐｅｔｉｔｉｖｅＥｘｔｒｏｇｅｎｉｃｐａｌｉｎｄｒｏｍｉｃｓｅｑｕｅｎｃｅＲＥＰ）进行了ＰＣＲ扩增和凝胶电泳,并对电泳结果的指纹图型进行了相似性聚类分析．结果表明了四川土著慢生花生根瘤菌的遗传多样性．指纹图形是根瘤菌多样性研究中一种简便而有效的方法     

Biological effect monitoring in dab (<Emphasis Type="Italic">Limanda limanda</Emphasis>) using gene transcript of CYP1A1 or EROD—a comparison

BACKGROUND, AIM, AND SCOPE: Gene expression analyses with real-time (RT)-polymerase chain reaction (PCR) gains importance in marine monitoring. This new technique has to be compared to the classical approaches like the well known biomarker ethoxyresorufin-O-deethylase (EROD) to test their suitability for monitoring programmes. The goal of the present study is to compare EROD activity and CYP1A1 mRNA expression in the important monitoring fish species dab (Limanda limanda) and to answer the question of whether these parameters reflect the polycyclic aromatic hydrocarbon (PAH) contamination of the fish. Further on, glyceraldehyd-3-phosphate dehydrogenase (GAPDH) was investigated as a potential housekeeping gene. MATERIALS AND METHODS: Female dab were caught in the summer of 2004 in the North Sea and in the Baltic. EROD activity was determined in liver samples by a kinetic fluorimetric assay according to a standard protocol. The gene expression of CYP1A (cytochrome P450 1A) and GAPDH were determined by means of RT-PCR. Results were compared to gonado somatic index and to the concentration of PAH metabolite 1OHPyr (1-hydroxypyrene) analysed in the bile fluids of the fish, respectively. RESULTS: Dab from all stations showed a considerable individual variation in the levels of both CYP1A mRNA and EROD. Highest mean values for CYP1A mRNA and EROD were detected in the northern part of the sampling area. In contrast, the PAH metabolite 1OHPyr was found at the highest concentration in fish caught near the German coast. CYP1A mRNA and EROD showed only a minor but significant correlation (r = 0.32, p < 0.05, n = 123). 1OHPyr in bile correlated significantly (p < 0.05) with the amount of GAPDH mRNA content in the liver. DISCUSSION: The significant but low correlation of CYP1A mRNA and EROD activity on an individual basis illustrates that these two parameters are apparently not closely linked. However, maximum EROD values correspond with maximum CYP1A mRNA concentrations when station means are regarded. Because EROD and CYP1A mRNA in dab follow different physiological principles, their application will lead to related but not identical monitoring results. This should be taken into account when future marine monitoring programmes are designed. The results also indicate that PAH are not the crucial factor for CYP1A and EROD levels in dab from the off-shore areas in the North Sea. This is remarkable because the PAH metabolism is known to be CYP1A-dependent and the widely used biomarker EROD has been recommended for monitoring PAH-related effects in fish from the North Sea. Due to a correlation between GAPDH and 1OHPyr, GAPDH was not suitable as housekeeping gene for dab. CONCLUSIONS: Neither the results from EROD nor from CYP1A1 mRNA measurements in dab reflected their exposure to PAH as measured by the PAH metabolite 1OHPyr. Thus, the question arises of whether EROD or CYP1A mRNA is a suitable biomarker at all to indicate PAH exposure in dab from the open North Sea. RECOMMENDATIONS AND PERSPECTIVES: For future biological effect monitoring, it is advisable to measure more and predominately independent parameters by RT-PCR and to incorporate more components of the detoxification system.     

铜绿假单胞菌增强型绿色荧光蛋白标记的研究

根据增强型绿色荧光蛋白(EGFP)基因设计了上下游引物P1和P2,通过对铜绿假单胞菌菌株X3绿色荧光蛋白标记研究,构建了铜绿假单胞菌标记系统,获得带有EGFP标记的基因工程菌X9,为进行相关的跟踪研究创造了条件.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,外源EGFP基因存在于转化子染色体上. 图 6参 14     

一种活性污泥总DNA提取方法的优化

对3种活性污泥DNA提取方法———传统蛋白酶K-SDS-氯仿异戊醇法(CPSCI法)、液氮研磨法和脱腐处理法进行了对比,并针对CPSCI法从污泥量、保温时间、裂解方式及沉淀时间等4个方面进行了优化。结果表明,优化的蛋白酶K-氯仿-异戊醇法(OPSCI法)采用污泥量0.10 g、37℃静置10 min,加SDS常温旋涡振荡及异丙醇直接离心等条件可获得长度在23.1 kb左右的DNA,OD260nm/OD280nm为1.86,稀释10倍后即可进行16S rDNA PCR。该方法重复性好,提取得率高,纯度好,操作简便,为常规实验室开展活性污泥微生物多样性研究提供了帮助。     

好氧堆肥高温期的嗜热真菌和嗜热放线菌群落结构

使用传统的培养方法和PCR-DGGE技术对好氧堆肥高温期的嗜热真菌和嗜热放线菌群落结构进行了研究.分别采用园林垃圾和餐厨垃圾作为堆肥原料,进行了20d好氧堆肥.高温期(≥50℃)持续了10d和8d.分别对2堆体高温期样品进行稀释平板混菌培养,真菌总数和放线菌总数均分别呈"降低-升高"和"升高-降低-升高"的趋势.同时提取微生物总DNA.分别使用真菌引物对(GC-NS7/NS8)和放线菌引物对(F243/GC-R513)从总DNA中成功扩增得到目标产物,对目标产物进行DGGE分离.传统培养法和DGGE图谱结果显示,不同堆体高温期的嗜热真菌和嗜热放线菌均表现出相似的变化规律,嗜热真菌优势菌比嗜热放线菌明显,但菌群总数比嗜热放线菌少.聚类分析结果表明,堆肥高温期嗜热真菌和嗜热放线菌分别以升温时56℃和58℃为界,分成2个明显的变化阶段,每阶段内部聚类关系较近.阶段间关系较远.温度对高温期真菌和放线菌具有明显的筛选作用.     

高氨氮废水与城市生活污水短程硝化系统菌群比较

短程硝化是污水脱氮工艺中的重要环节,系统中的菌群结构决定了其处理效果.为探讨短程硝化系统中的微生物对不同污水的适应性,利用细菌16S rDNA克隆文库、磷脂脂肪酸(PLFA)和定量PCR分析方法对高氨氮废水和城市生活污水短程硝化系统中活性污泥的细菌群落结构、总体微生物的多样性以及功能微生物进行了比较.克隆文库结果表明两个系统中细菌群落结构明显不同,城市生活污水中细菌种类更丰富,但两个系统的优势菌群都属于变形菌门(Proteobacteria)和拟杆菌门(Bacteroidete).磷脂脂肪酸分析结果显示高氨氮废水短程硝化系统中微生物多样性指数和均匀度指数明显低于城市生活污水.定量PCR结果表明,高氨氮废水短程硝化系统中氨氧化菌(AOB)与亚硝酸盐氧化菌(NOB)的数量都多于城市生活污水短程硝化系统;高氨氮废水短程硝化系统中AOB比NOB高3个数量级,而城市生活污水短程硝化系统中AOB比NOB高2个数量级.