应用FISH法对污水生物处理系统中细菌数量的计数

近年来,FISH(Fluorescence in Situ Hybridization,荧光原位杂交)法作为一种应用分子生物学技术对细胞及细菌等微生物进行定性以及定量分析的研究方法,在国际上得到广泛的应用。本文介绍了应用FISH法对污水处理工艺中细菌数量的计数方法。同时介绍了活性污泥试样制作、FISH法观测试样的制作以及各种试剂的配制、作用以及使用方法,基因探针的选择原则、杂交方法等实际操作方法与过程。     

2,4,6-Trinitrotoluene mineralization and bacterial production rates of natural microbial assemblages from coastal sediments

The nitrogenous energetic constituent, 2,4,6-Trinitrotoluene (TNT), is widely reported to be resistant to bacterial mineralization (conversion to CO₂); however, these studies primarily involve bacterial isolates from freshwater where bacterial production is typically limited by phosphorus. This study involved six surveys of coastal waters adjacent to three biome types: temperate broadleaf, northern coniferous, and tropical. Capacity to catabolize and mineralize TNT ring carbon to CO₂ was a common feature of natural sediment assemblages from these coastal environments (ranging to 270+/−38 μg C kg⁻¹ d⁻¹). More importantly, these mineralization rates comprised a significant proportion of total heterotrophic production. The finding that most natural assemblages surveyed from these ecosystems can mineralize TNT ring carbon to CO₂ is consistent with recent reports that assemblage components can incorporate TNT ring carbon into bacterial biomass. These data counter the widely held contention that TNT is recalcitrant to bacterial catabolism of the ring carbon in natural environments.     

Cellular uptake and mutagenic potential of metal oxide nanoparticles in bacterial cells

Extensive production and consumption of nanomaterials such as ZnO and TiO₂ has increased their release and disposal into the environment. The accumulation of nanoparticles (NPs) in ecosystem is likely to pose threat to non-specific targets such as bacteria. The present study explored the effect of ZnO and TiO₂ NPs in a model bacterium, Salmonella typhimurium. The uptake of ZnO and TiO₂ bare NPs in nano range without agglomeration was observed in S. typhimurium. TEM analysis demonstrated the internalization and uniform distribution of NPs inside the cells. Flow cytometry data also demonstrates that both ZnO and TiO₂ NPs were significantly internalized in the S. typhimurium cells in a concentration dependent manner. A significant increase in uptake was observed in the S. typhimurium treated even with 8 and 80 ng mL⁻¹ of ZnO and TiO₂ NPs with S9 after 60 min, possibly the formation of micelles or protein coat facilitated entry of NPs. These NPs exhibited weak mutagenic potential in S. typhimurium strains TA98, TA1537 and Escherichia coli (WP2uvrA) of Ames test underscoring the possible carcinogenic potential similar to certain mutagenic chemicals. Our study reiterates the need for re-evaluating environmental toxicity of ZnO and TiO₂ NPs presumably considered safe in environment.     

Microbial community response to addition of polylactate compounds to stimulate hexavalent chromium reduction in groundwater

To evaluate the efficacy of bioimmobilization of Cr(VI) in groundwater at the Department of Energy Hanford site, we conducted a series of microcosm experiments using a range of commercial electron donors with varying degrees of lactate polymerization (polylactate). These experiments were conducted using Hanford Formation sediments (coarse sand and gravel) immersed in Hanford groundwater, which were amended with Cr(VI) and several types of lactate-based electron donors (Hydrogen Release Compound, HRC; primer-HRC, pHRC; extended release HRC) and the polylactate-cysteine form (Metal Remediation Compound, MRC). The results showed that polylactate compounds stimulated an increase in bacterial biomass and activity to a greater extent than sodium lactate when applied at equivalent carbon concentrations. At the same time, concentrations of headspace hydrogen and methane increased and correlated with changes in the microbial community structure. Enrichment of Pseudomonas spp. occurred with all lactate additions, and enrichment of sulfate-reducing Desulfosporosinus spp. occurred with almost complete sulfate reduction. The results of these experiments demonstrate that amendment with the pHRC and MRC forms result in effective removal of Cr(VI) from solution most likely by both direct (enzymatic) and indirect (microbially generated reductant) mechanisms.     

Toxicity of imidazolium- and pyridinium-based ionic liquids and the co-metabolic degradation of N-ethylpyridinium tetrafluoroborate

We examined the effects of the ionic liquids (ILs), 1-butyl-3-methylimidazolium hexafluorophosphate [BMIM][PF₆], N-ethylpyridinium tetrafluoroborate [EtPy][BF₄], and N-ethylpyridinium trifluoroacetate [EtPy][CF₃COO] on Pseudomonas fluorescens, a ubiquitous soil bacterium. In the presence of 0.5- and 1% of [BMIM][PF₆] or [EtPy][CF₃COO] the growth of bacteria was inhibited, whereas exposing them to 1% [EtPy][BF₄] increased the lag period wherein bacteria adapt to growth conditions before continuing to grow. However, at higher concentrations (5% and 10%), no growth was observed. The inhibitory effects were evident by a decrease in the optical density of the culture, a decline in the consumption of the carbon source, citric acid, and a change in the size of the bacterium. At concentrations below 1%, [EtPy][BF₄] was metabolized by P. fluorescens in the presence of citric acid. Oxidation of the side alkyl-chain of [EtPy][BF₄] caused the accumulation of N-hydroxylethylpyridinium and pyridinium as major degradation products.     

硫化氢硫醇废气的臭氧氧化试验

介绍了臭氧氧化法去除工业废气中的硫化氢、硫醇的试验情况,考察了臭氧浓度、停留时间和催化作用等不同因素对去除率的影响.试验结果表明能达到较好的去除效果,硫化氢、硫醇的最终氧化产物不是二氧化硫,不会增加二氧化硫的污染.     

Effects of surfactants on the combined toxicity of TiO2 nanoparticles and cadmium to Escherichia coli

The combined ecological toxicity of TiO₂ nanoparticles (nano-TiO₂) and heavy metals has been paid more attention. As the common pollutants in water environment, surfactants could affect the properties of nanoparticles and heavy metals, and thus further influence the combined toxicity of nano-TiO₂ and heavy metals. In this study, the effects of sodium dodecyl benzene sulfonate (SDBS) and Tween 80 on the single and combined toxicities of Cd^2 + and nano-TiO₂ to Escherichia coli (E. coli) were examined, and the underlying influence mechanism was further discussed. The results showed both SDBS and Tween 80 enhanced the toxicity of Cd^2 + to E. coli in varying degrees. The reaction of SDBS and Cd^2 + could increase the outer membrane permeability and the bioavailability of Cd, while Tween 80 itself could enhance the outer membrane permeability. The combined toxicity of nano-TiO₂ and Cd^2 + to E. coli in absence of surfactant was antagonistic because of the adsorption of Cd^2 + to nano-TiO₂ particles. However, in the presence of SDBS, both SDBS and nano-TiO₂ influenced the toxicity of Cd^2 +, and also SDBS could adsorb to nano-TiO₂ by binding to Cd^2 +. The combined toxicity was reduced at Cd^2 + lower than 4 mg/L and enhanced at Cd^2 + higher than 4 mg/L under multiple interactions. Tween 80 enhanced the combined toxicity of nano-TiO₂ and Cd^2 + by increasing the outer membrane permeability. Our study firstly elucidated the effects of surfactants on the combined toxicity of nano-TiO₂ and Cd^2 + to bacteria, and the underlying influencing mechanism was proposed.     

Indoor and outdoor bioaerosol levels at recreation facilities, elementary schools, and homes

One major deficiency in linking environmental exposure to health effects is the current lack of data on environmental exposure. Therefore, to address this issue, the present study measured the bacterial and fungal concentrations in the indoor and outdoor air from two types of recreation facility (42 bars and 41 Internet cafes), 44 classrooms at 11 elementary schools, and 20 homes under uncontrolled environmental conditions during both summer and winter. No major environmental problems were reported at the four microenvironments being investigated during the entire study period. Bacteria and fungi were found in all the air samples, and the environmental occurrence of individual fungi was in the order of Cladosprium, Penicillium, Aspergillus, and Alternaria. The six parameters surveyed in the present study were all found to influence the indoor and outdoor bioaerosol levels: microenvironment type, sampling time in elementary school classrooms, agar type for measuring the fungal species, seasonal variation, facility location, and summer survey periods. The indoor and outdoor air concentrations of bacteria and fungi found in this study were comparable to those in other reports, with GM values for the total bacteria and total fungi between 10 and 10³ colony-forming units per cubic meter of air (CFU m⁻³). The fungal concentrations found at most of the indoor environments fell within the specified guidelines of the American Conference of Government Industrial Hygienists (ACGIH), between 100 and 1000 CFU m⁻³ for the total fungi. However, the indoor bioaerosol concentrations at most of the surveyed environments exceeded the Korean indoor bioaerosol guideline (800 CFU m⁻³). Consequently, the current findings suggest the need for reducing strategy for indoor microorganisms at the surveyed microenvironments.     

铬(Ⅵ)土著还原菌筛选及初步鉴定实验研究

以重铬酸钾为供试物,用固体平板法和还原实验对铬(Ⅵ)土著还原菌进行筛选,用载片培养法、革兰氏染色法、鞭毛染色法、芽孢染色法对还原菌做初步的鉴定,共筛选出16株铬(Ⅵ)土著还原菌,并挑选出还原率相对较高的Z2(35.2%)、Z3(45.2%)、Z4(38.6%)、X8(30.4%)、X10(29.4%)作为铬(Ⅵ)的优势还原菌株。经初步鉴定,土著真菌Z2为黑曲霉(Aspergillusniger),Z3、Z4为镰刀菌属(Fusariumsp.),土著细菌X8、X10为芽孢杆菌(Bacillussp.)。实验的方法和成果将为铬(Ⅵ)污染土壤微生物治理技术的推广应用提供技术支持。     

高效工程菌的构建及其降解有机农药的研究进展

综述了目前环境治理保护中构建高效工程菌常用的质粒转移、原生质体融合和基因重组等方法。介绍了工程菌在处理有机农药污染方面的降解性能及其固定化研究,并简要阐述了工程菌改造及应用方面所存在的问题及未来的研究展望。