硫自养高氯酸盐还原菌培养与驯化

考察了以污水处理厂的好氧污泥作为接种污泥,以S0作为电子供体培养驯化硫自养高氯酸盐还原菌的过程,并利用PCR-DGGE技术对不同阶段的污泥样品进行群落分析。结果表明,接种污泥在经过短暂的适应期后能具有较高的还原ClO-4的性能,随着驯化时间的延长,降解速率显著提高。培养驯化过程中微生物种群结构发生了演替变化,菌种Dechloromonas sp.CL、Quadrisphaera granulorum、Comamonadaceae bacterium 32-4、Acidovorax caeni、filamentous bacterium Plant1Iso8、Candidatus Nitrospira defluvii存在整个驯化培养阶段。随着培养驯化时间的延长,菌种Bacteroidetes bacterium S22-33、Herbaspirillum huttiense逐渐消失,出现新菌种Methyloversatilis universalis、beta proteobacterium HTCC379、beta proteobacterium CDB21和Clostridium bifermentans,并逐渐成为优势菌种。     

Kinetics for a membrane reactor reducing perchlorate.

The major objectives of this work were to operate and construct an autohydrogenotrophic reactor and estimate perchlorate degradation kinetics. The results show that autohydrogenotrophic bacteria were cultured in the reactor and capable of removing 3.6 mg/d of perchlorate in the presence of excess hydrogen (99% removal). The reactor was successful in treating the average influent perchlorate concentration of 532 microg/L to the level of 3 microg/L. A first-order relationship was obtained between the concentration of active biomass in the reactor and the hydraulic retention time for the given amount of substrate. During the kinetic loading study, perchlorate removal ranged from 100 to 50%. The kinetic rate of perchlorate degradation observed in this study was 1.62 hr(-1). The significant degradation of perchlorate in these samples indicates the ubiquity of perchlorate-reducing organisms. Additionally, nitrate was simultaneously removed during water treatment (greater than 90% removal). Because of the excess levels of hydrogen, simultaneous removal of nitrate was not believed to significantly affect perchlorate removal. The area of concern was the lack of complete control over biological treatment. The growth of sulfate-reducing organisms in the reactor negatively affected perchlorate removal efficiency. There were no significant effects observed on the dissolved organic carbon and total suspended solids concentration of the effluent, suggesting that the treatment did not produce a large amount of biomass washout.     

In situ bioremediation of nitrate and perchlorate in vadose zone soil for groundwater protection using gaseous electron donor injection technology.

When present in the vadose zone, potentially toxic nitrate and perchlorate anions can be persistent sources of groundwater contamination. Gaseous electron donor injection technology (GEDIT), an anaerobic variation of petroleum hydrocarbon bioventing, involves injecting electron donor gases, such as hydrogen or ethyl acetate, into the vadose zone, to stimulate biodegradation of nitrate and perchlorate. Laboratory microcosm studies demonstrated that hydrogen and ethanol promoted nitrate and perchlorate reduction in vadose zone soil and that moisture content was an important factor. Column studies demonstrated that transport of particular electron donors varied significantly; ethyl acetate and butyraldehyde were transported more rapidly than butyl acetate and ethanol. Nitrate removal in the column studies, up to 100%, was best promoted by ethyl acetate. Up to 39% perchlorate removal was achieved with ethanol and was limited by insufficient incubation time. The results demonstrate that GEDIT is a promising remediation technology warranting further validation.     

Bioremediation potential of a perchlorate-enriched sewage sludge consortium

The purpose of this work was to explore the reductive bioremediation potential of a perchlorate-enriched facultative anaerobic consortium. Rapid perchlorate reduction and bacterial growth were observed up to 1.84 g l(-1) of perchlorate, but not at 3.82 g l(-1) due to the toxicity. The specific growth rate of the mixed consortium was 0.1 h(-1). The consortium co-reduced perchlorate and nitrate with acetate as e- donor and carbon source. The presence of nitrate slowed down the perchlorate reduction rate. The other e- acceptors utilized include oxygen, chlorate, Cr(VI), and selenate. Over 95% of the 16 mg l(-1) of added Cr(VI) was reduced within 24 h of incubation with a high-density perchlorate-grown consortium. However, the consortium failed to couple growth with reduction of nitrite, sulfate, thiosulfate, and sulfite. During the search for autotrophic perchlorate reduction, many consortia from very diverse natural sources could not use sulfur compounds such as thiosulfate as e- donor.     

Isolation of a selected microbial consortium capable of degrading methyl parathion and p-nitrophenol from a contaminated soil site

A bacterial consortium with the ability to degrade methyl parathion and p-nitrophenol, using these compounds as the only carbon source, was obtained by selective enrichment in a medium with methyl parathion. Samples were taken from Moravia, Medellin; an area that is highly contaminated, owing to the fact that it was used as a garbage dump from 1974 to 1982. Acinetobacter sp, Pseudomonas putida, Bacillus sp, Pseudomonas aeruginosa Citrobacter freundii, Stenotrophomonas sp, Flavobacterium sp, Proteus vulgaris, Pseudomonas sp, Acinetobacter sp, Klebsiella sp and Proteus sp were the microorganisms identified within the consortium. In culture, the consortium was able to degrade 150 mg L?1 of methyl-parathion and p-nitrophenol in 120 h, but after adding glucose or peptone to the culture, the time of degradation decreased to 24 h. In soil, the consortium was also able to degrade 150 mg L?1 of methyl parathion in 120 h at different depths and also managed to decrease the toxicity.     

Symbiotic efficiency of autochthonous arbuscular mycorrhizal fungus (G. mosseae) and Brevibacillus sp. isolated from cadmium polluted soil under increasing cadmium levels

The effect of inoculation with indigenous naturally occurring microorganisms (an arbuscular mycorrhizal (AM) fungus and rhizosphere bacteria) isolated from a Cd polluted soil was assayed on Trifolium repens growing in soil contaminated with a range of Cd. One of the bacterial isolate showed a marked PGPR effect and was identified as a Brevibacillus sp. Mycorrhizal colonization also enhanced Trifolium growth and N, P, Zn and Ni content and the dually inoculated (AM fungus plus Brevibacillus sp.) plants achieved further growth and nutrition and less Cd concentration, particularly at the highest Cd level. Increasing Cd level in the soil decreased Zn and Pb shoot accumulation. Coinoculation of Brevibacillus sp. and AM fungus increased shoot biomass over single mycorrhizal plants by 18% (at 13.6 mg Cd kg(-1)), 26% (at 33.0 mg Cd kg(-1)) and 35% (at 85.1 mg Cd (kg(1)). In contrast, Cd transfer from soil to plants was substantially reduced and at the highest Cd level Brevibacillus sp. lowered this value by 37.5% in AM plants. Increasing Cd level highly reduced plant mycorrhization and nodulation. Strong positive effect of the bacterium on inocula, are important in plant Cd tolerance and development in Cd polluted soils.     

一株亚硝酸盐降解菌的分离鉴定与系统发育分析

从水产养殖污泥中分离筛选了一株优良的亚硝酸盐降解菌AQ-3,其在1.0×107 cfu/mL时对50mg/L亚硝酸盐的去除率高达99.47%。通过细菌自动鉴定系统(API ID32GN系统)以及16SrDNA系统发育分析,菌株AQ-3被鉴定为鲍曼氏不动杆菌(Acinetobacter baumannii)(JF751054),其16SrDNA序列与基因库中不动杆菌属菌株的16SrDNA序列有99%的同源性,而且与鲍曼氏不动杆菌菌株14(FJ907197)的亲缘关系最近。此外,菌株AQ-3同时含有nirS基因、nirK基因和norB基因,这些基因对AQ-3高效降解亚硝酸盐具有重要的意义。     

用于处理冲厕海水的固定化生物硅藻土小球的制备

选用聚乙烯醇（PVA）和海藻酸钠（SA）混合物作为包埋载体，对筛选出的耐盐复合菌群固定化制备方法进行了研究。通过正交实验分别研究了包埋载体不同配比、包菌量、硅藻土投加量和交联时间对固定化生物硅藻土小球的性能及其对冲厕污水中COD去除效果的影响。研究结果表明，固定化生物硅藻土小球最佳包埋条件是：聚乙烯醇9％，海藻酸钠0．5％，包菌量l：1，硅藻土20g／L，交联时间为24h。在该条件下，小球成球效果较好，机械强度高，对海水冲厕污水中COD的去除率达86．95％。     

Biodegradation of atrazine by Rhodococcus sp. BCH2 to N-isopropylammelide with subsequent assessment of toxicity of biodegraded metabolites

Atrazine is a persistent organic pollutant in the environment which affects not only terrestrial and aquatic biota but also human health. Since its removal from the environment is needed, atrazine biodegradation is achieved in the present study using the bacterium Rhodococcus sp. BCH2 isolated from soil, long-term treated with atrazine. The bacterium was capable of degrading about 75 % atrazine in liquid medium having pH 7 under aerobic and dark condition within 7 days. The degradation ability of the bacterium at various temperatures (20–60 °C), pH (range 3–11), carbon (glucose, fructose, sucrose, starch, lactose, and maltose), and nitrogen (ammonium molybdate, sodium nitrate, potassium nitrate, and urea) sources were studied for triumph optimum atrazine degradation. The results indicate that atrazine degradation at higher concentrations (100 ppm) was pH and temperature dependent. However, glucose and potassium nitrate were optimum carbon and nitrogen source, respectively. Atrazine biodegradation analysis was carried out by using high-performance thin-layer chromatography (HPTLC), Fourier transform infrared spectroscopy (FTIR), and liquid chromatography quadrupole time-of-flight (LC/Q-TOF-MS) techniques. LC/Q-TOF-MS analysis revealed formation of various intermediate metabolites including hydroxyatrazine, N-isopropylammelide, deisopropylhydroxyatrazine, deethylatrazine, deisopropylatrazine, and deisopropyldeethylatrazine which was helpful to propose biochemical degradation pathway of atrazine. Furthermore, the toxicological studies of atrazine and its biodegraded metabolites were executed on earthworm Eisenia foetida as a model organism with respect to enzymatic (SOD and Catalase) antioxidant defense mechanism and lipid peroxidation studies. These results suggest innocuous degradation of atrazine by Rhodococcus sp. BCH2 in nontoxic form. Therefore the Rhodococcus sp.BCH2 could prove a valuable source for the eco-friendly biodegradation of atrazine pesticide.     

Middle-thermophilic sulfur-oxidizing bacteria Thiomonas sp. RAN5 strain for hydrogen sulfide removal

Hydrogen sulfide (H2S) is one of the most toxic and offensively odorous gases and is generated in anaerobic bioreactors. A middle-thermophilic sulfur-oxidizing bacterium (SOB), Thiomonas sp. strain RAN5, was isolated and applied for H2S removal from both artificial and anaerobically digested gas. When a bioreactor containing medium inoculated with RAN5 was aerated continuously with artificial gas (containing 100 ppm H2S) at 45 degrees C for 156 hr, the H2S concentration in the vented gas was reduced by 99%. This was not affected by the presence of other microbes in the bioreactor The H2S removal efficiency of the RAN5 bioreactor for anaerobically digested gas was greater than 99% at influent H2S concentrations ranging from 2 to 1800 ppm; the efficiency decreased to 90% at influent H2S concentrations greater than 2000 ppm. Thiomonas sp. strain RAN5 cannot survive at room temperature, and thus its leakage from a wastewater treatment plant would not damage sewage systems. These data suggest that Thiomonas sp. strain RAN5 may be a useful microorganism for H2S removal.     

Characterisation and optimisation of three potential aerobic bacterial strains for kraft lignin degradation from pulp paper waste

Eight aerobic bacterial strains were isolated from pulp paper mill effluent sludge. Out of eight through nutrient enrichment technique three potential aerobic bacterial strains ITRC S(6), ITRC S(7) and ITRC S(8) were found capable to effectively degrade the kraft lignin (KL), a major byproduct of the chemical pulping process and main contributor to the colour and toxicity of effluent. Further, these potential strains (ITRC S(6), ITRC S(7) and ITRC S(8)) were biochemically characterised as Gram variable small rod, Gram negative rod and Gram positive rod respectively. Subsequently, 16S rRNA sequencing showed 95% base sequence homology and it was identified as Paenibacillus sp. (AY952466), Aneurinibacillus aneurinilyticus (AY856831), Bacillus sp. (AY952465) for ITRC S(6), IITRC S(7) and ITRC S(8), respectively. In batch decolourization experiments Bacillus sp. ITRC S(8) reduced the colour of lignin amended mineral salt medium, pH 7.6 by 65% after 6th d, at 30 degrees C, A. aneurinilyticus ITRC S(7) by 56% and Paenibacillus ITRC S(6) 43%. Under these conditions the three strains degraded the KL by 37%, 33% and 30%, respectively while the mixed culture of these three bacteria reduced colour by 69%, lignin by 40% and total substrate by 50% under same conditions. Biodegradation of the KL was not affected by low (<0.2 mg l(-1)) dissolved oxygen content; thus oxygen inhibition is more likely to be a metabolism-dependent event. Initially with 48 h incubation the decolourization was slow with decreased pH. Further incubation there was rapid decolourization with slight increase in pH at 6d compared with initial pH by increasing culture optical density. The lignin analysis from medium with HPLC indicated complete degradation rather than biotransformation with complete loss of absorbance peak at 280 nm.     

Brine reuse in ion-exchange softening: salt discharge, hardness leakage, and capacity tradeoffs

Ion-exchange water softening results in the discharge of excess sodium chloride to the aquatic environment during the regeneration cycle. In order to reduce sodium chloride use and subsequent discharge from ion-exchange processes, either brine reclaim operations can be implemented or salt application during regeneration can be reduced. Both result in tradeoffs related to loss of bed volumes treated per cycle and increased hardness leakage. An experimentally validated model was used to compare concurrent water softening operations at various salt application quantities with and without the direct reuse of waste brine for treated tap water of typical midwestern water quality. Both approaches were able to reduce salt use and subsequent discharge. Reducing salt use and discharge by lowering the salt application rate during regeneration consequently increased hardness leakage and decreased treatment capacity. Single or two tank brine recycling systems are capable of reducing salt use and discharge without increasing hardness leakage, although treatment capacity is reduced.     

Mineralisation of 2,6-dichlorobenzamide (BAM) in dichlobenil-exposed soils and isolation of a BAM-mineralising Aminobacter sp

Mineralisation of the groundwater contaminant 2,6-dichlorobenzamide (BAM), a metabolite from the herbicide 2,6-dichlorobenzonitrile (dichlobenil), was studied in soil samples obtained from 39 locations previously exposed to dichlobenil. Rapid BAM mineralisation was detected in samples from six locations with 5.2-64.6% of the added BAM mineralised within 48-50 days. From one location rapid BAM mineralisation was observed in soil samples down to a depth of 2 m below the surface. One location with fast BAM mineralisation showed significant dichlobenil degradation activity with 25.5% of the added dichlobenil being mineralised within 50 days. By inoculating soil showing the fastest mineralisation of BAM into a mineral medium with BAM as the only carbon and nitrogen source an enrichment culture was established. Community analysis based on extracted DNA revealed a change of the bacterial community but without any clear indication of key members within the BAM-mineralising culture. Parallel cultivation resulted for the first time in the isolation of a BAM-mineralising bacterium, identified as an Aminobacter sp.     

氯氰菊酯降解菌CY22-7的分离鉴定及降解特性研究

从采集的多种土壤样品中.获得了19株在3种筛选平板上均生长良好的候选氯氰菊酯降解菌.通过生理生化特性鉴定和16S rDNA序列的测定及比对.将其中的氯氰菊酯降解菌CY22-7鉴定为中华根瘤菌属(Sinorhizobium sp.).同时,对氯氰菊酯降解菌CY22-7的降解特性研究结果表明:(1)氯氰菊酯降解菌CY22-7以5%(体积分数)的接种鼍接种到氯氰菊酯起始质量浓度为100 mg/L的氯氰菊酯乙醇培养基后,于200 r/min、30℃摇床中培养.经过6 d的培养,氯氰菊酯降解菌CY22-7降解了约60%的氯氰菊酯.(2)加人外源营养物质有利于促进氯氰菊酯的降解.其中,葡萄糖和酵母提取物的促进作用最为明显.(3)氯氰菊酯降解菌CY22-7降解氯氰菊酯的最适温度为30℃,最佳pH为6.0.     

Isolation and characterization of mesotrione-degrading Bacillus sp. from soil

Dissipation kinetics of mesotrione, a new triketone herbicide, sprayed on soil from Limagne (Puy-de-Dôme, France) showed that the soil microflora were able to biotransform it.Bacteria from this soil were cultured in mineral salt solution supplemented with mesotrione as sole source of carbon for the isolation of mesotrione-degrading bacteria. The bacterial community structure of the enrichment cultures was analyzed by temporal temperature gradient gel electrophoresis (TTGE). The TTGE fingerprints revealed that mesotrione had an impact on bacterial community structure only at its highest concentrations and showed mesotrione-sensitive and mesotrione-adapted strains. Two adapted strains, identified as Bacillus sp. and Arthrobacter sp., were isolated by colony hybridization methods.Biodegradation assays showed that only the Bacillus sp. strain was able to completely and rapidly biotransform mesotrione. Among several metabolites formed, 2-amino-4-methylsulfonylbenzoic acid (AMBA) accumulated in the medium. Although sulcotrione has a chemical structure closely resembling that of mesotrione, the isolates were unable to degrade it.     

氮源对克雷伯氏菌NⅢ2分泌絮凝剂特性的影响

实验研究了蔗糖为碳源,硝酸钠、脲、蛋白胨、硫酸铵和氯化铵等氮源对NⅢ2发酵产絮凝剂的影响。结果表明,发酵液起始pH值为7.50,以硝酸钠为氮源,发酵液pH会上升,升至7.60～8.34时,NⅢ2菌株开始大量分泌微生物絮凝剂,发酵72 h,产量可达7.5 g/L,该产量是目前报道的克雷伯氏菌产絮凝剂的最高值。脲为氮源,pH则下降,降至5.04～6.49时,大量分泌絮凝剂,发酵72 h产量达5.2 g/L。蛋白胨、氯化铵和硫酸铵等为氮源时,pH下降十分明显,pH小于3.71时有絮凝剂分泌,发酵72 h产量约2.0 g/L或更小。以硝酸钠和脲为氮源时,发酵液中有黄色物质分泌,该黄色物质出现或黄色逐渐加深,是NⅢ2菌高产絮凝剂的标志。除硫酸铵外,其他氮源发酵所产絮凝剂为O-糖蛋白。当以硝酸钠、脲、蛋白胨、硫酸铵和氯化铵为氮源时,絮凝剂中蛋白的含量分别为9.55%、33.28%、19.39%、13.81%和15.51%,且蛋白含量越高,絮凝剂活性越大。     

脱氮副球菌YF1微生物燃料电池生物阴极脱氮和产电

以脱氮副球菌YF1构建纯种生物阴极微生物燃料电池(microbial fuel cell,MFC)进行脱氮和产电机理的研究。研究结果发现,阴极碳氮比、pH值对产电和脱氮效率有明显影响。当MFC的阴极运行条件pH值为8.0,碳氮比为20时,运行时间15 h时,脱氮率高达100%,输出电压为150 mV。上述结果表明,微生物燃料电池运行过程中,细菌降解硝酸根的机理为将硝酸根还原为N2或者直接将其作为自身的营养物质而利用。循环伏安(CV)与扫描电镜(SEM)的结果表明,在微生物燃料电池运行中,副球菌YF1通过接触导电作为产电的电子供体。     

A long-term performance test on an autotrophic denitrification column for application as a permeable reactive barrier

The long-term performance of a sulfur-based reactive barrier system was evaluated using autotrophic denitrification in a large-scale column. A bacterial consortium, containing autotrophic denitrifiers attached on sulfur particles, serving as an electron donor, was able to transform 60mgNL(-1) of nitrate into dinitrogen. In the absence of phosphate, the consortium was unable to remove nitrate, but after the addition of phosphate, nitrate removal was readily evident. Once the column operation had stabilized, seepage velocities of 1.0x10(-3) and 0.5x10(-3)cms(-1), corresponding to hydraulic residence times of 24 and 48h, respectively, did not affect the nitrate removal efficiency, as determined by the nitrate concentration in the effluent. However, data on the nitrate, nitrite and sulfate distribution along the column indicated differential transformation patterns with column depths. Based on the dinitrogen concentration in the total gas collected, the denitrification efficiency of the tested column was estimated to be more than 95%. After 500d operation, the hydrodynamic characteristics of the column slightly changed, but these changes did not inhibit the nitrate removal efficiency. Data from a bacterial community analysis obtained from four parts of the column demonstrated the selective a spatial distribution of predominant species depending on available electron acceptors or donors.     

Biological denitrification in drinking water treatment using the seaweed Gracilaria verrucosa as carbon source and biofilm carrier.

Chemical and microbiological aspects were investigated with regard to biological denitrification of drinking water using the seaweed Gracilaria verrucosa as the carbon and energy substrate and as physical support for the microbial flora in semibatch, fixed-bed reactors. Complete removal of nitrate (100 mg/L) was readily achieved without accumulation of nitrite. Microbiological analysis indicated that the effluent of the reactor contained high numbers of bacteria (>10(6)/mL total count). Among the 44 bacterial strains isolated directly from the samples or isolated after enrichment at 37 degrees C, 25 different fatty acid profiles were found, indicating a complex microflora, including potential pathogens.     

Tolerance to trichlorophenols in microorganisms from a polluted and a pristine site of a river

The effect of 2,4,5- and 2,4,6-trichlorophenol on the microbiota from a polluted and a pristine site of a river was studied. Bacterial metabolic activity measurements by epifluorescence microscopy showed that the polluted site contained more metabolically active cells than the pristine site. Total culturable bacterial counts and tolerant bacterial counts from both sites were not affected by incubation (for up to 5 days) with 200 ppm of chlorophenols. However, the incubation with 500 ppm of 2,4,5-trichlorophenol prevented detection of total and tolerant bacterial counts in the pristine site, and inhibited tolerants in the polluted site. None of 250 bacterial colonies directly isolated from these samples was able to grow on chlorophenols. However, bacteria able to grow on 2,4,6-trichlorophenol, were obtained by enrichment of water and sediments samples.