Reductive dissolution of Pu(IV) by Clostridium sp. under anaerobic conditions

An anaerobic, gram positive, spore-forming bacterium Clostridium sp., common in soils and wastes, capable of reduction of Fe(III) to Fe(II), Mn(IV) to Mn(II), Tc(VII) to Tc(IV), and U(VI) to U(IV), reduced Pu(IV) to Pu(III). Addition of 242Pu (IV)-nitrate to the bacterial growth medium at pH 6.4 resulted in the precipitation of Pu as amorphous Pu(OH)4 due to hydrolysis and polymerization reactions. The Pu (1 x 10(-5) M) had no effect upon growth of the bacterium as evidenced by glucose consumption; carbon dioxide and hydrogen production; a decrease in pH of the medium from 6.4 to 3.0 due to production of acetic and butyric acids from glucose fermentation; and a change in the Eh of the culture medium from +50 to -180 mV. Commensurate with bacterial growth, Pu was rapidly solubilized as evidenced by an increase in Pu concentration in solution which passed through a 0.03 microm filtration. Selective solvent extraction of the culture by thenoyltrifluoroacetone (TTA) indicated the presence of a reduced Pu species in the soluble fraction. X-ray absorption near edge spectroscopic (XANES) analysis of Pu in the culture sample at the Pu LIII absorption edge (18.054 keV) showed a shift of -3 eV compared to a Pu(IV) standard indicating reduction of Pu(IV) to Pu(III). These results suggestthat, although Pu generally exists as insoluble Pu(IV) in the environment, under appropriate conditions, anaerobic microbial activity could affect the long-term stability and mobility of Pu by its reductive dissolution.     

Inactivation of Ascaris suum and poliovirus in biosolids under thermophilic anaerobic digestion conditions

There is considerable interest in the United States in production of Class A (low pathogen content) biosolids from the treatment of municipal wastewater sludge. Current requirements imposed by the U.S. Environmental Protection Agency make it difficult for thermophilic anaerobic digestion, in its simplest process configurations, to achieve Class A status. In particular, the time-temperature requirements necessitate long batch treatment times at temperatures associated with thermophilic anaerobic digestion. The time-temperature requirements are meant to ensure extensive inactivation of helminth eggs and enteric viruses, considered to be the most heat-resistant of the relevant pathogen classes. However, data on inactivation kinetics of these pathogens at precisely controlled and well-characterized temperatures are scarce. We measured inactivation of vaccine-strain poliovirus and eggs from the helminth Ascaris suum at temperatures from 49 to 55 degrees C in a lab-scale batch reactor containing biosolids from a continuous-flow thermophilic anaerobic digester. Both microbes were inactivated rapidly, with Ascaris more resistant to inactivation than poliovirus, and the relationships between inactivation rate and temperature were steep. The Arrhenius correlation between inactivation rate and temperature over the range 49-53 degrees C is consistent with protein denaturation as the inactivation mechanism for both microbes. The least stringent of the EPA time-temperature equations for thermal processes requires batch treatment times more than 2 orders of magnitude greater than would be required for three-log reduction of Ascaris at the rates we measured, suggesting an overly conservative regulatory approach. Such a grossly conservative approach can hinder full-scale implementation of thermophilic anaerobic digestion.     

The anaerobic conversion of methanol under thermophilic conditions: pH and bicarbonate dependence

The thermophilic (55 degrees C) anaerobic conversion of methanol was studied in an unbuffered medium (pH 4+/-0.2) and in a phosphate buffered medium (pH 6.4+/-0.1), in both cases without bicarbonate addition. Our cultivated sludge consortium was unable to degrade methanol under acidic conditions. During the 160 d of continuous operation of an up-flow anaerobic sludge blanket (UASB) reactor (R1), at an organic loading rate (ORL) of 6 gCOD/(l.d) and pH around 4, only 5% of the applied methanol load was consumed and no methane (CH4) was detected. However, hydrogenotrophic methanogens were found to be resistant to exposure to such conditions. At the end of the trial, the hydrogenotrophic methanogenic activity of the sludge was 1.23+/-0.16 gCOD/(gVSS.d) at neutral pH. With methanol as the test substrate, the addition of bicarbonate led to acetate accumulation. A second reactor (R2) was operated for 303 d at OLRs ranging from 5.5 to 25.4 gCOD/(l.d) in order to assess the conversion of methanol at neutral pH (phosphate buffered) in a bicarbonate deprived medium. The reactor performance was poor with a methanol-COD removal capacity limited to about 9.5 gCOD/(l.d). The system appeared to be quite susceptible to any type of disturbance, even at low OLR. The fraction of methanol-COD converted to CH4 and acetate was found to be unaffected by the OLR applied. At the end of the trial, the outcome of the competition was about 50% methanogenesis and 50% homoacetogenesis.     

糖蜜废水在两相厌氧消化工艺中的应用

针对糖蜜废水排放量大、有机负荷高、悬浮物浓度高和成分复杂等特点,结合两相厌氧消化工艺具有产酸相与产甲烷相分离的特性,在处理高浓度有机废水和能源回收利用上有独特优势进行讨论。并例举了糖蜜废水在两相厌氧消化工艺中的应用实例和特性研究,指出了两相厌氧消化工艺存在的问题和未来的发展趋势。     

Apparent thermal resistance of Bacillus stearothermophilus spores recovered under anaerobic conditions

 The effect of recovering Bacillus stearothermophilus spores under anaerobic conditions on their apparent thermal resistance was studied. Spores were suspended in bidistilled water as a reference medium, heated at 115, 117, 119, 121, 123 and 125°C and recovered under aerobic and anaerobic conditions. D values (decimal reduction time) obtained following recovery under anaerobic conditions were lower than those obtained under aerobic conditions. Reductions of between 31 and 48% were found for all the temperatures studied. When spores were suspended in mushroom extract and recovered under anaerobic conditions the apparent heat resistance was much lower than that obtained under aerobic conditions (D _121°C was 4.3 min and 1.7 min, under aerobic and anaerobic conditions, respectively). Heating the spores in mushroom extract and recovering the spores under anaerobic conditions produced an additive effect, decreasing the apparent heat resistance of the B. stearothermophilus spores. Received: 1 July 1997     

Apparent thermal resistance of Bacillus stearothermophilus spores recovered under anaerobic conditions

The effect of recovering Bacillus stearothermophilus spores under anaerobic conditions on their apparent thermal resistance was studied. Spores were suspended in bidistilled water as a reference medium, heated at 115, 117, 119, 121, 123 and 125°C and recovered under aerobic and anaerobic conditions. D values (decimal reduction time) obtained following recovery under anaerobic conditions were lower than those obtained under aerobic conditions. Reductions of between 31 and 48% were found for all the temperatures studied. When spores were suspended in mushroom extract and recovered under anaerobic conditions the apparent heat resistance was much lower than that obtained under aerobic conditions (D _121°C was 4.3 min and 1.7 min, under aerobic and anaerobic conditions, respectively). Heating the spores in mushroom extract and recovering the spores under anaerobic conditions produced an additive effect, decreasing the apparent heat resistance of the B. stearothermophilus spores.     

一株高产Mn-SOD菌发酵条件的优化

对一株产Mn-SOD的耐碱芽孢杆菌Bacillus sp.110-3的发酵条件进行了优化.通过对培养基碳源组成,氮源组成,碳氮比例以及培养基始初pH和金属离子对菌株Bacillus sp.110-3产Mn-SOD的影响进行研究,确定了培养基的最佳组成为:玉米粉4%,蛋白胨1%,酵母膏0.5%,氯化钠1%,Mn2100μmol/L,pH6.0～8.0.应用正交试验设计结合SPSS软件分析,确定了各因素对产酶影响的显著程度,为实际生产中最佳工艺的确定提供了理论基础.     

糖蜜发酵裂殖壶菌生产二十二碳六烯酸

对糖蜜作为碳源发酵裂殖壶菌生产二十二碳六烯酸(docosahexaenic acid,DHA)进行了初步研究:选择不同来源的甘蔗糖蜜与粗糖蜜培养裂殖壶菌生产DHA,考察了糖蜜来源、糖蜜添加比例对裂殖壶菌发酵生物量、菌体油脂含量与DHA含量的影响。结果表明,甘蔗糖蜜中的杂质(灰分、胶体)对菌体生长与油脂积累存在明显的抑制作用,即使大幅降低糖蜜添加比例效果也不理想,限制了其作为替代碳源在实际生产上的应用。在此基础上考察了杂质含量相对较低的粗糖蜜的情况,在添加比例低于80%的情况下,其培养效果与对照葡萄糖非常接近,可以部分替代葡萄糖用于裂殖壶菌发酵生产。     

利用UASB反应器厌氧发酵糖厂废水生物制氢的研究进展

介绍了糖厂废水利用上流式厌氧污泥床（UASB）反应器进行厌氧发酵制氢的原理、工程控制参数,重点阐述了其研究现状和发展趋势.利用UASB反应器进行生物制氢,在理论研究方面已取得许多成果,但仍然没有实现大规模工业化生产,因为在启动和运行中仍存在一些问题,本文提出了一些解决策略和建议,以期对今后的研究有一定的指导意义.     

一株产中温淀粉酶芽孢杆菌的培养条件优化

从土壤中分离到1株产淀粉酶的芽孢杆菌,通过单因素和正交试验对其产酶条件进行了优化。结果表明,该菌株在1.5%玉米淀粉、1%牛肉膏、0.15%硫酸镁、0.1%氯化钙培养基中,接种量为2%（v/v）、初始pH值为7.0、装液量10%、发酵温度37℃、180r/min振荡培养24h时酶活力最高,其酶活达到了2155.45U/mL。     

Purification and characterization of an extracellular protease from alkaliphilic and thermophilic Bacillus sp. PS719

An alkaline protease was purified to apparent homogeneity from culture supernatants of Bacillus sp. PS719, a novel alkaliphilic, thermophilic bacterium isolated from a thermal spring soil sample, by ammonium sulfate precipitation followed by DEAE-cellulose and alpha-casein agarose column chromatographies. The purified enzyme migrated as a single protein band of 42 kDa during both denaturing and nondenaturing gel electrophoresis, suggesting that it consists of a single polypeptide chain. Its isoelectric point was approximately 4.8. The protease exhibited maximum activity towards azocasein at pH 9.0 and at 75 degrees C. The enzyme activity was stimulated by Ca2+, but was inhibited in the presence of Fe2+ or Cu2+. The enzyme was stable in the pH range 8.0 to 10.0 and up to 80 degrees C in the absence of Ca2+. Since phenylmethylsulfonyl fluoride (PMSF) and 3,4-dichloroisocoumarin (DCI) in addition to N-alpha-p-tosyl-L-lysine chloromethyl ketone (TLCK) completely inhibited the activity, this enzyme appears to be a trypsin-like serine protease. Among the various oligopeptidyl-p-nitroanilides tested, the protease showed a preference for cleavage at arginine residues on the carboxylic side of the scissile bond of the substrate, liberating p-nitroaniline from N-carbobenzoxy (CBZ)-L-arginine-p-nitroanilide with the K(m) and V(max) values of 0.6 mM and 1.0 micromol.min(-1).mg protein(-1), respectively.     

Thermostable collagenolytic activity of a novel thermophilic isolate, Bacillus sp. strain NTAP-1

We isolated an acidophilic thermophile belonging to the genus Bacillus, strain NTAP-1, which secreted a thermostable collagenolytic activity into the culture medium. The collagenolytic activity exhibited an optimum pH for Azocoll hydrolysis of pH 3.9 and was not completely inhibited by 10 mM ethylenediaminetetraacetic acid (residual activity, 63%), suggesting that Bacillus NTAP-1 produces a novel acid proteinase with highest activity for collagen. The collagenolytic activity was thermostable; more than 80% of the original activity was retained after incubation of the culture supernatant at pH 4.0 and 60 degrees C for 4 h.     

嗜热菌Geobacillus sp.PZH1产木聚糖酶发酵条件的优化

对嗜热菌Geobacillus sp.PZH1发酵产嗜热耐碱木聚糖酶的培养条件进行了优化研究。对碳源、氮源、初始pH、接种量以及发酵温度五个因素进行了单因素实验,在此基础上对碳氮比、初始pH以及接种量进行了正交实验。结果表明,该菌株在发酵培养7d时有最大产酶量,Geobacillus sp.PZH1发酵产木聚糖酶最佳发酵条件为:桦木木聚糖为碳源,牛肉膏为氮源,碳氮比2∶3,初始pH7.0,接种量4%,发酵温度50℃,发酵时间7d。在最佳产酶条件下进行发酵,木聚糖酶活力可达2.56IU/mL,是未优化前酶活的1.44倍。     

糖蜜酒精废液的脱色实验研究

以糖蜜酒精废液为研究对象,探讨糖蜜酒精废液的脱色处理.实验先用聚合氯化铝(PAC)和聚丙烯酰胺(PAM)以5∶1复配对糖蜜酒精进行絮凝预处理,可使浊度下降95%,COD去除90%,色度也有一定的下降.然后以活性炭、强氯精及活性炭-强氯精复配对废液进行脱色研究,结果表明单独采用活性炭吸附,脱色效果最好,且在投加量为34 g/L,pH值为4,反应时间为180min,脱色率达95%.     

利用甘蔗糖蜜厌氧发酵产丁二酸的研究

对甘蔗糖蜜作为廉价碳源厌氧发酵制备丁二酸进行初步研究.实验证明,Actinobacillus succinogenes NJ113能够利用葡萄糖、果糖、蔗糖等碳源,制备丁二酸为菌体利用廉价甘蔗糖蜜作碳源进行丁二酸发酵提供可行性依据.通过比较不同的糖蜜预处理方法,得出经硫酸处理的糖蜜发酵效果最好,丁二酸浓度37.73g/L,比未经处理的糖蜜所产丁二酸浓度高12.6%.考察不同的糖蜜添加量对发酵结果的影响表明,初始总糖浓度为65g/L时,丁二酸的产量最高为49.63g/L;在3L罐中进行放大实验,丁二酸产量46.91g/L,质量收率为72.2%,分别比混合糖(含蔗糖、果糖、葡萄糖)的发酵结果高9.8%、10%.     

Removal of estrogens in municipal wastewater treatment under aerobic and anaerobic conditions: consequences for plant optimization

The removal of estrogens (estrone E1, estradiol E2, and ethinylestradiol EE2) was studied in various municipal wastewater treatment processes equipped for nutrient removal. A biological degradation model is formulated, and kinetic parameters are evaluated with batch experiments under various redox conditions. The resulting model calculations are then compared with sampling campaigns performed on differenttypes of full-scale plant: conventional activated-sludge treatment, a membrane bioreactor, and a fixed-bed reactor. The results show a > 90% removal of all estrogens in the activated sludge processes. (Due to the analytical quantification limit and low influent concentrations, however, this removal efficiency represents only an observable minimum.) The removal efficiencies of 77% and > or = 90% for E1 and E2, respectively, in the fixed-bed reactor represent a good performance in view of the short hydraulic retention time of 35 min. The first-order removal-rate constant in batch experiments observed for E2 varied from 150 to 950 d(-1) for a 1 gSS L(-1) sludge suspension. The removal efficiency of E1 and EE2 clearly depends on the redox conditions, the maximum removal rate occurring under aerobic conditions when E1 was reduced to E2. Sampling campaigns on full-scale plants indicate that the kinetic values identified in batch experiments (without substrate addition) for the natural estrogens may overestimate the actual removal rates. Although this paper does not give direct experimental evidence, it seems that the substrate present in the raw influent competitively inhibits the degradation of E1 and E2. These compounds are therefore removed mainly in activated sludge compartments with low substrate loading. Theoretical evaluation leads us to expect that diffusive mass transfer inside the floc (but not across the laminar boundary layer) appreciably influences the observed degradation rates of E1 and E2, but not of EE2.     

芽孢杆菌Bacillus sp.nov.SK006产纤溶酶发酵条件的研究

利用实验室自行从亚洲传统发酵食品虾酱中筛选到一株产纤溶酶能力较强的芽孢杆菌(Bacillus sp.nov.SK006)进行发酵,考察培养基组成及培养条件对该菌种产纤溶酶能力的影响.结果表明该菌株产酶最佳的组分(质量浓度)为:葡萄糖20 g/L,胰蛋白胨30 g,L,Na2HPO4·12H2012 g/L,NaH1PO4.2H20 1.3 g/L,Mgs04·7H20 1.0 g/L;培养条件:种龄18 h,接种量3%,发酵周期24 h,初始pH为7.0,摇床转速180 r/min,发酵温度37℃,在此条件下发酵液纤溶活性(以Plasmin为标准)达2.63 U/mL,优化后纤维蛋白溶解酶的产量是优化前0.70 U/mL的3.76倍.     

绿色木霉发酵强化糖蜜酒精废液活性炭脱色效果的研究

采用绿色木霉菌株对糖蜜酒精废液进行生物处理,并对处理后的废液进行活性炭脱色试验.考察不同活性炭吸附处理条件和不同绿色木霉培养条件对糖蜜酒精废液生物法脱色的影响,并进行了正交实验.结果表明,在绿色木霉接种量为1%(v/v),培养时间为36h,培养温度为26℃,活性炭投加量为5%的条件下糖蜜酒精废液脱色效果最好,透光率达91.57%.     

Anaerobic treatment performance and microbial population of thermophilic upflow anaerobic filter reactor treating awamori distillery wastewater

Distillery wastewater from awamori making was anaerobically treated for one year using thermophilic upflow anaerobic filter (UAF) reactors packed with pyridinium group-containing nonwoven fabric material. The microbial structure and spatial distribution of microorganisms on the support material were characterized using molecular biological methods. The reactor steadily achieved a high TOC loading rate of 18 g/l/d with approximately 80% TOC removal efficiency when non-diluted wastewater was fed. The maximum TOC loading rate increased to 36 g/l/d when treating thrice-diluted wastewater. However, the TOC removal efficiency and gas evolution rate decreased compared with that when non-diluted wastewater was used. Methanogens closely related to Methanosarcina thermophila and Methanoculleus bourgensis and bacteria in the phyla Firmicutes and Bacteroidetes were predominant methanogens and bacteria in the thermophilic UFA reactor, as indicated by 16S rRNA gene clone analysis. Fluorescence in situ hybridization (FISH) results showed that a large quantity of bacterial cells adhered throughout the whole support, and Methanosarcina-like methanogens existed mainly in the relative outside region while Methanoculleus cells were located in the relative inner part of the support. The support material used proved to be an excellent carrier for microorganisms, and a UAF reactor using this kind of support can be used for high-rate treatment of awamori/shochu distillery wastewater.     

糖蜜酒精废水处理方法的研究

综述了糖蜜酒精废水的治理技术,分析了农田灌溉法、浓缩处理法、厌氧法,包括UASB、IC、ABR、UMAR、好氧法、厌氧—好氧处理法、氧化处理法,包括WAO、SCWO、其他方法,如EM、微氧技术的优缺点.认为资源化结合治理方法是处理糖蜜酒精废水的发展方向.