Studies of the Stability of Microbial Association Use in Industrial Biofiltering of Gaseous Discharges

Results of industrial exploitation of a biofiltration plant tailored for purifying gaseous discharges of hazardous organic components such as toluene, cyclohexane, and xylene, are examined. Both numerical and compositional variations were monitored for a long-term (more than 1.5 years) utilization process in an association of microorganisms decomposing organic pollutants. A population of microbial association composed by one yeast and two bacterial strains in the biofilm on the surface of filtering sheets was abundant (10⁸–10⁹ yeast cells/cm² and 10¹⁰–10¹¹ bacterial cells/cm²) and stable during the whole period of monitoring. A microbial association in the culture medium averaging 10⁶ yeast cells/l and 10⁸ bacterial cells/l is more susceptible to technogenic impacts and seasonal fluctuations. Overall, the biofilter as an open and autonomic system maintained its microbial association, thereby providing high-degree (93–98%) purification of industrial gaseous discharges from organic pollutants.     

Bacterial degradation of airborne phenol in the phyllosphere

Despite the vast surface area of terrestrial plant leaves and the large microbial communities they support, little is known of the ability of leaf-associated microorganisms to access and degrade airborne pollutants. Here, we examined bacterial acquisition and degradation of phenol on leaves by an introduced phenol degrader and by natural phyllosphere communities. Whole-cell gfp-based Pseudomonas fluorescens bioreporter cells detected phenol on leaves that had previously been transiently exposed to gaseous phenol, indicating that leaves accumulated phenol; moreover, they accumulated it in sites that were accessible to epiphytic bacteria and to concentrations that were at least 10-fold higher than those in the air. After inoculated leaves were exposed to gaseous 14C-phenol, leaves harbouring the phenol-degrading Pseudomonas sp. strain CF600 released eight times more 14CO2 than did leaves harbouring a non-degrading mutant, demonstrating that CF600 actively mineralized phenol on leaves. We evaluated phenol degradation by natural microbial communities on green ash leaves that were collected from a field site rich in airborne organic pollutants. We found that significantly more phenol was mineralized by these leaves when the communities were present than by these leaves following surface sterilization. Thus, phenol-degrading organisms were present in these natural communities and were metabolically capable of phenol degradation. Collectively, these results provide the first direct evidence that bacteria on leaves can degrade an organic pollutant from the air, and indicate that bacteria on leaves could potentially contribute to the natural attenuation of organic air pollutants.     

Microbial community structure and biogeochemistry of Miocene subsurface sediments: implications for long-term microbial survival

Thirty closely spaced cores were obtained from Miocene-aged fluvial, lacustrine and palaeosol subsurface sediments ranging in depth from 173 to 197 m at a site in south-central Washington to investigate the size and composition of the microbial community in relation to sediment geochemical and geophysical properties. Total phospholipid fatty acid (PLFA) analysis indicated that the greatest concentrations of microbial bio-mass were in low-permeability lacustrine sediments that also contained high concentrations of organic carbon. Community structure, based on lipid analyses and on in situ hybridization of bacterial cells with 16S RNA-directed DNA probes, also revealed the presence of metabolically active bacteria that respire sulphate and/or Fe(III) in the lacustrine sediments. Concentrations of pore water sulphate were low (4–8 mg/L) and HCI-extractable Fe was predominantly Fe(II) in the same samples where total biomass and organic carbon were highest. The low hydraulic conductivity (10^-6 to < 10^-9 cm/s) of these sediments has likely contributed to the long term maintenance of both bacteria and organic carbon by limiting the supply of soluble electron acceptors for microbial respiration. These results suggest that the current subsurface microbial population was derived from organisms that were present during lake sedimentation = 6–8 million years ago.     

中亚热带地区两种森林植被类型土壤微生物群落结构

利用磷脂脂肪酸(PLFA)生物标记法分析了中亚热带地区罗浮栲天然林和相邻的杉木人工林土壤微生物群落结构特点.结果表明: 两种植被类型的磷脂脂肪酸总量、细菌特征脂肪酸、真菌特征脂肪酸、放线菌特征脂肪酸、革兰氏阳性菌和革兰氏阴性菌特征脂肪酸含量均为0～10 cm高于10～20 cm土层,罗浮栲天然林高于杉木人工林.在两种植被类型的两个土层中,细菌PLFAs含量均显著高于真菌PLFAs含量.两种植被类型中,细菌PLFAs含量约占PLFAs总量的44%～52%,而真菌仅占6%～8%,表明细菌在该地区两种植被类型土壤中处于优势地位.主成分分析表明,土壤微生物群落结构差异主要由植被类型差异引起,土层深度的影响相对较小.相关分析显示,革兰氏阴性菌、革兰氏阳性菌以及细菌的PLFAs含量与pH呈显著负相关,与含水量呈显著正相关;土壤微生物主要类群PLFAs含量与总氮、有机碳、C/N和铵态氮均呈显著正相关.     

The use of co-immobilization of Trichosporon cutaneum and Bacillus licheniformis for a BOD sensor

The microorganisms Trichosporon cutaneum and Bacillus licheniformis were used to develop a microbial biochemical oxygen demand (BOD) sensor. It was found that T. cutaneum gave a greater response to glucose, whereas B. licheniformis gave a better response to glutamic acid. Hence, co-immobilized T. cutaneum and B. licheniformis were used to construct a glucose and glutamic acid sensor with improved sensitivity and dynamic range. A membrane loading of T. cutaneum at 1.1x10(8 )cells ml(-1) cm(-2) and B. licheniformis at 2.2x10(8) cells ml(-1) cm(-2) gave the optimum result: a linear range up to 40 mg BOD l(-1) with a sensitivity of 5.84 nA mg(-1) BOD l. The optimized BOD sensor showed operation stability for 58 intermittent batch measurements, with a standard deviation of 0.0362 and a variance of 0.131 nA. The response time of the co-immobilized microbial BOD sensor was within 5-10 min by steady-state measurement and the detection limit was 0.5 mg BOD l(-1). The BOD sensor was insensitive to pH in the range of pH 6.8-7.2.     

Isolation of an aboriginal bacterial community capable of utilizing cyanide, thiocyanate, and ammonia from metallurgical plant wastewater

An aboriginal bacterial community capable of degrading cyanide (10 mg/l) and thiocyanate (2 g/l) and eliminating ammonia (120 mg/l) had been isolated from recycled water samples after blast-furnace gas purification of a metallurgical plant wastewater. It was shown that the optimal conditions for this bacterial community were as follows: temperature, 34 degrees C; pH, 8.8-9.0; available organic matter concentration (glucose equivalent), 5 g/l; and dissolved O2 concentration, 8-10 mg/l. This aboriginal community was formed by the bacteria belonging to the genus Pseudomonas.     

Isolation, identification and application of novel bacterial consortium TJ-1 for the decolourization of structurally different azo dyes

A novel bacterial consortium (TJ-1), which could decolorize Acid Orange 7 (AO7) and manyother azo dyes, was developed. In TJ-1 three bacterial strains were identified as Aeromonas caviae, Proteus mirabilis and Rhodococcus globerulus by 16S rRNA gene sequence analysis. AO7 decolorization was significantly higher with the use of consortium as compared to the use of individual strains, indicating complementary interactions among these strains. AO7 decolorization was observed under microaerophilic condition in the presence of organic carbon source. Either yeast extract (YE) alone or a combination of YE and glucose resulted in much higher decolorization of AO7 as compared to glucose alone, peptone or starch. Kinetic studies with different initial AO7 concentrations showed that more than 90% decolorization could be achieved even at 200mg/l within 16h. Fed-batch studies showed that AO7 decolorization required 10h during the first cycle and 5h in the second and third cycles, showing that bacterial cells could be used for multiple cycles. The consortium also decolorized fifteen other azo dyes individually as well as a simulated wastewater containing a mixture of all the sixteen azo dyes, thus, conferring the possibility of application of TJ-1 for the treatment of industrial wastewaters.     

叶际微生物研究进展

植物的叶际是一个复杂的生态系统,微生物的生存环境条件严苛。其可被利用的营养成分较少,温湿度波动大。此外,较强的紫外线辐射对于叶际微生物的生存也有很大影响。但是植物叶际却有着丰富的微生物多样性,其中还有许多有益细菌和真菌。它们通过和植物寄主的互作,改善着叶际微生物的栖居环境;其对植物病原体的拮抗亦可提高植物的抗病性。植物叶际的微生物还可以产生激素以促进植物生长,还有一些微生物可以利用农药等污染有机物作为营养物质,在污染物的环境生物修复方面显示巨大的潜力。此外,叶际微生物作为一种生态学指标在生态稳定与环境安全评价中开始发挥显著的作用。     

Composition of organic matter in sediments facing a river estuary (Tyrrhenian Sea): relationships with bacteria and microphytobenthic biomass

The relationships between the biochemical composition of sediment organic matter and bacteria and microphytobenthic biomass distribution, were investigated along the coast of Northern Tuscany (Tyrrhenian Sea). Organic matter appeared to be of highly refractory composition. Among the three main biochemical classes, proteins were the major component (0.96 mg g^-1 sediment d.w.) followed by total carbohydrates (0.81 mg g^-1 sediment d.w.) and lipids (8.1 µg g^-1 sediment d.w.). Bacterial number in surface sediments (0–2 cm) ranged from 1.7 to 24.5 × 10⁸ cells g^-1 of sediment dry weight showing a strong decrease with sediment depth. In surface sediments, significant correlations were found between bacterial biomass and protein concentration. Bacterial activity (measured by the frequency of dividing cells) was significantly related to lipid concentration. Bacterial and microphytobenthic biomass accounted for 3.1 and 18.1% respectively of the sediment organic carbon. In surface sediments bacterial lipids accounted, on average, for 27 % of total lipids, whereas bacterial proteins and carbohydrates accounted for 2.5 and 0.5% of total proteins and carbohydrates, respectively.The benthic degradation process indicated that lipids were a highly degradable compound (about 35% in the top 10 cm). Carbohydrate decreased for 25.6% in the top 10 cm, whereas proteins increased with depth, thus indicating that this compound may resist to diagenetic decomposition.These data suggest that specific organic compounds need to be measured rather than bulk carbon and nitrogen measurements in order to relate microbial biomass to the quality of organic matter.     

氮肥添加对高寒藏嵩草(Kobresia tibetica)沼泽化草甸和土壤微生物群落的影响

以藏嵩草沼泽化草甸为研究对象,利用磷脂脂肪酸(PLFA)技术,研究连续6年N素添加对地上植被群落数量特征、土壤微生物群落结构的影响。结果表明:①藏嵩草沼泽化草甸群落生物量、枯枝落叶对施肥处理无明显响应,且莎草科植物对土壤氮素的吸收和利用率较低。②施肥增加了0-10 cm土壤微生物类群PLFAs丰富度尤其细菌和革兰氏阳性菌PLFAs,降低了10-20 cm PLFAs丰富度;③磷脂脂肪酸饱和脂肪酸/单烯不饱和脂肪酸、细菌PLFAs/真菌PLFAs的比值随土壤层次增加而增加;④0-10 cm土层革兰氏阳性菌、真菌PLFAs含量与pH、土壤速效磷、速效氮、土壤有机质显著正相关(P0.05或P0.01);10-20 cm土层,细菌、革兰氏阳性菌、真菌和总PLFAs含量与土壤有机质含量显著正相关(P0.05或P0.01)。表明藏嵩草沼泽化草甸微生物PLFAs含量和丰富度对施肥的响应存在明显的土层梯度效应,土壤微生物PLFAs含量和丰富度主要受表层土壤初始养分含量的影响。     

Inhibition of microbial adhesion to silicone rubber treated with biosurfactant from Streptococcus thermophilus A

Microbial adhesion of four bacterial and two yeast strains isolated from explanted voice prostheses to silicone rubber before and after conditioning with a biosurfactant obtained from the probiotic bacterium Streptococcus thermophilus A was investigated in a parallel plate flow chamber. The silicone rubber with and without an adsorbed biosurfactant layer was characterized using contact angle measurements. Water contact angles indicated that the silicone rubber surface with adsorbed biosurfactant was more hydrophilic (58 degrees) than bare silicone rubber (109 degrees). The results obtained showed that the biosurfactant was effective in decreasing the initial deposition rates, and the number of bacterial cells adhering after 4 h, for all microorganisms tested. A decrease in the initial deposition rate was observed for Rothia dentocariosa GBJ 52/2B and Staphylococcus aureus GB 2/1 from 1937+/-194 to 179+/-21 microorganisms cm(-2) s(-1) and from 1255+/-54 to 233+/-26 microorganisms cm(-2) s(-1), respectively, accounting for an 86% reduction of the initial deposition rate for both strains. The number of bacterial cells adhering to the silicone rubber with preadsorbed biosurfactant after 4 h was further reduced by 89% and 97% by the two strains, respectively. The two yeast strains tested showed less reduction in adhesion after 4 h, to values between 67% and 70%. Such a pretreatment with surface-active compounds may constitute a promising strategy to reduce the microbial colonization rate of silicone rubber voice prostheses.     

Bacterial dominance in subseafloor sediments characterized by methane hydrates

The degradation of organic carbon in subseafloor sediments on continental margins contributes to the largest reservoir of methane on Earth. Sediments in the Andaman Sea are composed of ~ 1% marine-derived organic carbon and biogenic methane is present. Our objective was to determine microbial abundance and diversity in sediments that transition the gas hydrate occurrence zone (GHOZ) in the Andaman Sea. Microscopic cell enumeration revealed that most sediment layers harbored relatively low microbial abundance (10(3)-10(5) cells cm(-3)). Archaea were never detected despite the use of both DNA- and lipid-based methods. Statistical analysis of terminal restriction fragment length polymorphisms revealed distinct microbial communities from above, within, and below the GHOZ, and GHOZ samples were correlated with a decrease in organic carbon. Primer-tagged pyrosequences of bacterial 16S rRNA genes showed that members of the phylum Firmicutes are predominant in all zones. Compared with other seafloor settings that contain biogenic methane, this deep subseafloor habitat has a unique microbial community and the low cell abundance detected can help to refine global subseafloor microbial abundance.     

Degradation of waste waters from olive oil mills by Yarrowia lipolytica ATCC 20255 and Pseudomonas putida

Waste waters from olive oil processing may cause severe pollution in the Mediterranean area, since they have a high level of chemical oxygen demand (COD) (100–200 g/l) and contain other organic and inorganic compounds. In all olive oil producing countries, the reduction of pollution in olive oil mill waste waters at reasonable costs and using techniques suitable for most industrial applications is an unsolved problem. For this paper, the yeast Yarrowia lipolytica ATCC 20255 was grown on waste waters from an olive oil mill in a 3.5 1 fermenter under batch culture conditions. The results showed that the yeast was capable of reducing the COD value by 80% in 24 h. In this way, a useful biomass of 22.45 g/l as single cell protein (SCP) and enzyme lipase were produced. During this process, most of the organic and inorganic substances were consumed, only aromatic pollutants were still present in the fermentation effluents. Therefore, we used a phenol degrader, namely Pseudomonas putida, to reduce phenolic compounds in the fermentation effuents after removing Yarrowia lipolytica cells. P. putida was effective in reducing phenols in only 12 h.     

Physical and chemical factors affecting microbial biomass and activity in contaminated subsurface riverine sediments

Over 80 years of direct discharge of industrial effluents into the Mahoning River, located in northeastern Ohio, USA, has led to the accumulation of a wide variety of pollutants within its sediments. This study examined the physical and chemical parameters, including lipophilic pollutants, affecting microbial activity and biomass in subsurface (10-40 cm horizon) sediments. Microbial biomass was higher in anthropogenically contaminated sediments, and step-wise linear regression showed that approximately 82% of the variation in microbial biomass could be explained by total hexane extractable hydrocarbons, sediment particle size, and water content. There was no correlation between microbial activity and biomass. Independent variables influencing anaerobic activity were temperature and water holding capacity. The results of this study indicate that freshwater, sedimentary anaerobic microbial communities respond to a range of environmental parameters, many of which influence subsurface river sediments, and that lipophilic pollutants, when present, can cause increases in total microbial biomass.     

瓦屋山扁刺栲-中华木荷常绿阔叶次生林土壤有机碳组分特征

次生林在全球碳循环中占有重要地位,为了研究中国中亚热带次生林土壤有机碳组分特征,以四川瓦屋山中山段扁刺栲-中华木荷常绿阔叶次生林为对象,通过挖取土壤剖面分层(0—10、10—40、40—70 cm和70—100 cm)取样方式,研究土壤各有机碳组分特征。结果表明:土壤有机碳、微生物生物量碳、可浸提溶解性有机碳和易氧化碳含量均随土层深度增加而减小,0—10 cm土层有机碳含量为121.89 g/kg,高于已报道的亚热带其他常绿阔叶林和四川各类森林;0—10 cm层微生物生物量碳含量为1931.82 mg/kg,可浸提溶解性有机碳含量为697.42 mg/kg,易氧化碳含量为20.98 g/kg,高于已报道的许多相似天然林和人工林活性碳含量。土壤有机碳储量为154.87 t/hm2,在四川省各类森林中处于中等水平。研究表明瓦屋山扁刺栲-中华木荷常绿阔叶次生林活性碳含量较大,微生物活动和养分流动较为活跃,凋落物层转化为土壤碳的潜力较大,这类生态系统可能会在区域碳循环过程中扮演更为重要的角色。     

Biodegradation of xylene and butyl acetate using an aqueous-silicon oil two-phase system

A stable microbial population, consisting of seven bacterial strains and three yeast strains, was selected in batch cultures on a mixture of ortho and meta-xylene and butyl acetate as the sole source of carbon and energy. This population can completely degrade up to 10 g/L of a mixture of these xenobiotics (70% xylene and 30% butyl acetate wt/wt) in a two-phase aqueous-silicone oil system (70%/30% vol/vol) within 96 h, while for the usual one-phase system very low growth degradation rates were observed. Further organic solvents were tested and finally, silicon oil was selected as the best organic phase for such a two-phase system. With periodical pH adjustments to 6.0 in fed-batch mode, the culture showed a global degradation rate of 63 mg L^-1 h^-1.     

Control of Lactobacillus contaminants in continuous fuel ethanol fermentations by constant or pulsed addition of penicillin G

The addition of penicillin G to combat microbial contamination in continuous fuel alcohol fermentations was performed using both continuous and pulsed addition regimes. In continuous fermentations where both Saccharomyces cerevisiae and Lactobacillus paracasei were present, the mode of addition of penicillin G determined final numbers of viable L. paracasei. When the same overall average concentration of penicillin G was added in both pulsed and continuous modes, the initial viable number of L. paracasei (8.0 x 10(9) cfu ml(-1)) decreased to a greater degree (1.02 x 10(5) cfu ml(-1) L. paracasei) when penicillin G was pulsed at 6 h frequencies at an overall average concentration of 2,475 U/l than when penicillin G was added continuously at 2,475 U/l (2.77 x 10(5) cfu ml(-1) L. paracasei). Pulsed additions over longer frequencies at 2,475 U/l were not as effective in reducing viable bacteria. Viable yeasts increased during both treatment conditions by more than 2-fold. The two addition regimes also eliminated the 40% decrease in ethanol concentration caused by the intentional bacterial infection. Although there was 3 times more bacterial death with 6 h pulsed additions compared to continuous additions of penicillin G at 2,475 U/l, there was, by that point, no practical difference in either final ethanol concentration or relative ethanol recovery.     

Measurement of biologically available naphthalene in gas and aqueous phases by use of a Pseudomonas putida biosensor

Genetically constructed microbial biosensors for measuring organic pollutants are mostly applied in aqueous samples. Unfortunately, the detection limit of most biosensors is insufficient to detect pollutants at low but environmentally relevant concentrations. However, organic pollutants with low levels of water solubility often have significant gas-water partitioning coefficients, which in principle makes it possible to measure such compounds in the gas rather than the aqueous phase. Here we describe the first use of a microbial biosensor for measuring organic pollutants directly in the gas phase. For this purpose, we reconstructed a bioluminescent Pseudomonas putida naphthalene biosensor strain to carry the NAH7 plasmid and a chromosomally inserted gene fusion between the sal promoter and the luxAB genes. Specific calibration studies were performed with suspended and filter-immobilized biosensor cells, in aqueous solution and in the gas phase. Gas phase measurements with filter-immobilized biosensor cells in closed flasks, with a naphthalene-contaminated aqueous phase, showed that the biosensor cells can measure naphthalene effectively. The biosensor cells on the filter responded with increasing light output proportional to the naphthalene concentration added to the water phase, even though only a small proportion of the naphthalene was present in the gas phase. In fact, the biosensor cells could concentrate a larger proportion of naphthalene through the gas phase than in the aqueous suspension, probably due to faster transport of naphthalene to the cells in the gas phase. This led to a 10-fold lower detectable aqueous naphthalene concentration (50 nM instead of 0.5 micro M). Thus, the use of bacterial biosensors for measuring organic pollutants in the gas phase is a valid method for increasing the sensitivity of these valuable biological devices.     

亚热带树种转换对林地土壤微生物群落结构和功能的影响

通过分析杉木采伐迹地营造阔叶树种尾巨桉和固氮树种黑木相思人工林后土壤微生物群落组成和酶活性,探讨造林树种转换对于改善杉木林地土壤微生物特性的影响.结果表明: 树种转换对土壤微生物群落组成和酶活性的影响主要局限于0～10 cm土壤层.杉木转换为固氮树种黑木相思后,显著提高了0～10 cm土壤层总脂肪酸含量、真菌、革兰氏阳性细菌、革兰氏阴性细菌和放线菌生物量.主成分分析表明,黑木相思人工林土壤微生物群落组成与杉木和尾巨桉人工林具有显著差异,土壤中革兰氏阳性细菌、阴性细菌和放线菌丰度显著提高.在0～10 cm土壤层,黑木相思人工林土壤纤维素水解酶、乙酰氨基-葡萄糖苷酶和酸性磷酸酶活性均显著高于杉木和尾巨桉人工林.研究表明,杉木转变为固氮树种黑木相思后会显著提高微生物生物量和酶活性,有助于土壤有机质的恢复,加快养分循环过程.     

Activity,Distribution, and Diversity of Sulfate Reducers and Other Bacteria in Sediments above Gas Hydrate (Cascadia Margin,Oregon)

Cold seep environments such as sediments above outcropping hydrate at Hydrate Ridge (Cascadia margin off Oregon) are characterized by methane venting, high sulfide fluxes caused by the anaerobic oxidation of methane, and the presence of chemosynthetic communities. Recent investigations showed that another characteristic feature of cold seeps is the occurrence of methanotrophic archaea, which can be identified by specific biomarker lipids and 16S rDNA analysis. This investigation deals with the diversity and distribution of sulfate-reducing bacteria, some of which are directly involved in the anaerobic oxidation of methane as syntrophic partners of the methanotrophic archaea. The composition and activity of the microbial communities at methane vented and nonvented sediments are compared by quantitative methods including total cell counts, fluorescence in situ hybridization (FISH), bacterial production, enzyme activity, and sulfate reduction rates. Bacteria involved in the degradation of particulate organic carbon (POC) are as active and diverse as at other productive margin sites of similar water depths. The availability of methane supports a two orders of magnitude higher microbial biomass (up to 9.6 2 10 10 cells cm m 3 ) and sulfate reduction rates (up to 8 w mol cm m 3 d m 1 ) in hydrate-bearing sediments, as well as a high bacterial diversity, especially in the group of i -proteobacteria including members of the branches Desulfosarcina/Desulfococcus , Desulforhopalus , Desulfobulbus , and Desulfocapsa . Most of the diversity of sulfate-reducing bacteria in hydrate-bearing sediments comprises seep-endemic clades, which share only low similarities with previously cultured bacteria.