Decolourization of azo dye methyl red by Saccharomyces cerevisiae MTCC 463

Saccharomyces cerevisiae MTCC 463 decolourizes toxic azo dye, methyl red by degradation process. Methyl red (100mgl(-1)) is degraded completely within 16min in plain distilled water under static anoxic condition, at the room temperature. Effect of physicochemical parameters (pH of medium, composition of medium, concentration of cells, concentration of dye, temperature and agitation) on methyl red decolourization focused the optimal condition required for decolourization. Biodegradation (fate of metabolism) of methyl red in plain distilled water was found to be pH dependent. Cells of Saccharomyces cerevisiae could degrade methyl red efficiently up to 10 cycles in plain distilled water. Analysis of samples extracted with ethyl acetate from decolourized culture flasks in plain distilled water (pH 6.5) and at pH 9 using UV-VIS, TLC, HPLC and FTIR confirm biodegradation of methyl red into several different metabolites. A study of the enzymes responsible for the biodegradation of methyl red in the control and cells obtained after decolourization in plain distilled water (pH 6.5) and at pH 9 showed different levels of the activities of laccase, lignin peroxidase, NADH-DCIP reductase, azoreductase, tyrosinase and aminopyrine N-demethylase. A significant increase in the activities of lignin peroxidase and NADH-DCIP reductase was observed in the cells obtained after decolourization in plain distilled water (pH 6.5), however cells obtained at pH 9 shows increased activities of azoreductase, tyrosinase, lignin peroxidase and NADH-DCIP reductase. High efficiency to decolourize methyl red in plain distilled water and low requirement of environmental conditions enables this yeast to be used in biological treatment of industrial effluent containing azo dye, methyl red.     

Reduction of selenite to red elemental selenium by moderately halotolerant Bacillus megaterium strains isolated from Bhitarkanika mangrove soil and characterization of reduced product

Two Gram (+) bacterial strains, BSB6 and BSB12, showing resistance and potential for Se(IV) reduction among 26 moderately halotolerant isolates from the Bhitarkanika mangrove soil were characterized by biochemical and 16S rDNA sequence analyses. Both of them were strictly aerobic and able to grow in a wide range of pH (4-11), temperature (4-40 °C) and salt concentration (4-12%) having an optimum growth at 37 °C, pH ∼7.5 and 7% salt (NaCl). The biochemical characteristics and 16S rDNA sequence analysis of BSB6 and BSB12 showed the closest phylogenetic similarity with the species Bacillus megaterium. Both the strains effectively reduced Se(IV) and complete reduction of selenite (up to 0.25 mM) was achieved within 40 h. SEM with energy dispersive X-ray and TEM analyses revealed the formation of nano size spherical selenium particles in and around the bacterial cells which were also supported by the confocal micrograph study. The UV-Vis diffuse reflectance spectra and XRD of selenium precipitates revealed that the selenium particles are in the nanometric range and crystalline in nature. These bacterial strains may be exploited further for bioremediation process of Se(IV) at relatively high salt concentrations and green synthesis of selenium nanoparticles.     

硝化菌的分离及特性分析

从联合脱硫脱氮填料塔中的陶粒上分离出两株硝化菌,命名为NS2和NS5,这2种菌均为革兰氏阴性菌。硝化能力测试结果表明:NS2和NS5能利用亚硝酸盐作为氮源,将亚硝酸盐转化成硝酸盐,硝化速率都达到60%以上。部分长度的16S rDNA序列同源性分析表明,NS2和NS5与Delftia sp.的同源性高达99%,并用MEGA程序对该菌株进行了系统发育进化分析。结合形态观察和生理生化鉴定,初步鉴定NS2,NS5为戴尔福特菌(Delftia sp.)。     

Influence of redox mediators and metal ions on synthetic acid dye decolourization by crude laccase from Trametes hirsuta

In this paper, the effect of redox mediators on synthetic acid dye decolourization (Sella Solid Red and Luganil Green) by laccase from Trametes hirsuta cultures has been investigated. All the redox mediators tested, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 1-hydroxybenzotriazole (HBT) and Remazol Brilliant Blue R (RBBR), led to higher activities than those obtained without mediators addition showing the suitability of the laccase/mediator system (LMS) in the decolourization of acid dyes. HBT was by far the most effective mediator, showing a decolourization percentage of 88% in 10 min for Sella Solid Red and of 49% in 20 min for Luganil Green. On the other hand, the stability of laccase against several metal ions, normally found in textile wastewater, was assessed. Laccase was stable at a concentration of 1mM for 7d against all the metal ions tested except for Zn+2, CrO4(-2), Cd+2, Cr2O7(-2), Fe+2, Cu+2 and especially Hg+2. When the concentration was increased to 10mM laccase stability decreased against all the metals assayed, in particular against Fe+2. In addition, the effect of metal ions on the decolourization process was also studied. It was found that Hg+2 inhibited the dye decolourization process, being the presence of HBT absolutely required for dye decolourization.     

Decolourization and removal of some organic compounds from olive mill wastewater by advanced oxidation processes and lime treatment

Background Olive mill wastewater (OMW) generated by the olive oil extracting industry is a major pollutant, because of its high organic load and phytotoxic and antibacterial phenolic compounds which resist biological degradation. Mediterranean countries are mostly affected by this serious environmental problem since they are responsible for 95% of the worldwide oliveoil production. There are many methods used for OMW treatment, such as adsorption, electro coagulation, electro-oxidation, biological degradation, advanced oxidation processes (AOPs), chemical coagulation, flocculation, filtration, lagoons of evaporation and burning systems, etc. Currently, there is no such economical and easy solution. The aim of this study was to evaluate the feasibility of decolourization and removal of phenol, lignin, TOC and TIC in OMW by UV/H₂O₂ (AOPs). The operating parameters, such as hydrogen peroxide dosage, times, pH, effect of UV and natural sunlight were determined to find the suitable operating conditions for the best removal. Moreover, there is no study reported in the literature related to the use of UV/H₂O₂ and lime together in OMW treatment. Methods OMW was obtained from an olive-oil producing plant (Muğla area of Turkey) which uses a modern production process. No chemical additives are used during olive oil production. This study was realised by using two different UV sources, while taking the time and energy consumption into consideration. These two sources were mercury lamps and natural sunlight. Before starting AOPs experiments, one litre of OMW was treated by adding lime until a pH of 7.00. Then, 100 ml was taken from each sample, and 1 to 10 ml of a 30% H₂O₂ (Riedel-deHaen) solution was added. These solutions in closed vessels were laid in the natural sunlight for a week and their compositions and colour changes were analysed daily by UV-Vis spectrophotometer. At the end of the one-week period, they were treated with lime. In this study, the effect of changes in the initial pH, times and H₂O₂ concentrations on removal was investigated. At the end of all experiments, changes in colour, phenol, lignin, TOC and TIC concentrations were analysed according to standard methods. Results and Discussion In the samples exposed to natural sunlight and having an H₂O₂/OMW ratio of 3 ml/100 ml, a significant colour removal was achieved approximately 90% of the time at the end of 7 days. When the same samples were treated with lime (pH: up to 7), 99% efficiency was achieved. When phenol and lignin removals were examined in the same concentration, phenol and lignin removal were found 99.5%, 35%, respectively. However, for maximum lignin removal, more use of H₂O₂ (10 ml H₂O₂/100 ml OMW) was found to be necessary. Under these conditions, it was found that lignin can be removed by 70%, but to 90% with lime, at the end of a seven-day period. Rate constants obtained in the experiments performed with direct UV were found to be much higher than those of the samples exposed to natural sunlight (k^a _lignin = 0.3883 ≫ k^b _lignin = 0.0078; k^a _phenol = 0.5187 ≫ k^b _phenol = 0.0146). Moreover, it should be remembered in this process that energy consumption may induce extra financial burden for organisations. Conclusions It was found, in general, that colour, lignin, total organic carbon and phenol were removed more efficiently from OMW by using H₂O₂ UV and lime OMW. Moreover, in the study, lime was found to contribute, both initially and after radical reactions, to the efficiency to a great extent. Recommendations and Perspectives Another result obtained from the study is that pre-purification carried out with hydrogen peroxide and lime may constitute an important step for further purification processes such as adsorption, membrane processes, etc.     

白腐菌J-6对五类化学结构染料的脱色性能

从白腐菌中筛出了一株能对五类不同化学结构的染料进行高效脱色的菌株——J-6。通过采用“批式”摇床培养法,测定染料脱色率,从而确定该菌株能高效脱色的最佳培养时间和连续处理染料的能力。发酵培养96h的菌体可使活性艳蓝R(A)SP在5h内脱色率达95％以上;活性翠蓝G、活性黑GRP、混合染料在9h内脱色率达90％以上;孔雀绿、Po1yR-478在24h内脱色率分别达95％、90％以上。另外,J-6对不同染料最佳脱色时间和最终脱色效率与菌体本身发酵培养的时间长短和染料的种类紧密相关。J6处理能力强,连续处理4次染料后仍保持较高的脱色率。     

Biodegradation of azo dyes by the yeast Candida zeylanoides in batch aerated cultures

A number of simple azo dyes was degraded in liquid aerated batch cultures by a strain of the yeast Candida zeylanoides. The standard decolorization medium contained glucose as a carbon and energy source, and its pH was either controlled to 5.0-5.2, or allowed to decrease to 3.2-2.8, in the course of microorganism growth. The extent of colour removal in the culture medium was assessed through the decrease in dye absorbance of the supernatants. The extent of colour removal ranged from 44 to 90%, after 7 days, for 5 out of 6 dyes studied in shaked cultures, without pH control, and from 46 to 67%, after 22 hours, for 6 out of 8 dyes in batch experiments, at controlled pH.     

不适宜生长条件对混茵降解苯胺的影响

分别从台州和衢州某化工厂的好氧池中分离筛选得到2株苯胺降解菌TZl和JH1,经16SrDNA测序鉴定为Comamonassp．TZ1和Pseudomonassp．JH1,均具有较强的苯胺降解能力,培养24h后,可使初始浓度为800mg／L的苯胺去除率达到96．4％～98．4％。在此基础上,按体积比l：1将2株菌液进行混合构建了混合菌体系,进而对比考察了苯胺初始浓度、pH、盐度和重金属等环境因子对单一菌和混合菌生长量及降解苯胺效果的影响,重点探讨混合菌对不适宜生长环境的适应性及其对苯胺的降解特性。通过单一菌和混合菌对比实验发现,在适宜苯胺初始浓度、pH和盐度条件下,混合菌的生长量略高于单一菌;在不适宜生长的高浓度苯胺、pH和盐度条件下,混合菌也表现出了更强的适应性和苯胺矿化能力。Zn2＋和Cr6＋耐受实验则表明,对于Cr6＋混合菌表现出了更强的耐受能力,而对于zn2＋并没有表现出更强的耐受能力。     

Decolourization of synthetic dyes by Trametes hirsuta in expanded-bed reactors

The present paper studies the decolourization of different synthetic dyes (Indigo Carmine, Bromophenol Blue, Methyl Orange and Poly R-478) by the white-rot fungus Trametes hirsuta at bioreactor scale under solid-state conditions, operating with ground orange peelings as a support-substrate. Dye decolourization was performed in both batch and continuous mode. Batch cultivation led to high decolourization percentages in a short time (100% for Indigo Carmine in 3h and 85% for Bromophenol Blue in 7 h). As for continuous cultivation, different hydraulic retention times (HRT) were studied (0.8, 1, 1.5 and 3d). The highest decolourization percentages were obtained operating at a HRT of 3d, especially for the dyes Methyl Orange and Poly R-478 (81.4% and 46.9%, respectively). This is a very interesting result, since there are few studies dealing with the continuous decolourization of dyes at bioreactor scale by fungal laccases.     

不适宜生长条件对混菌降解苯胺的影响

分别从台州和衢州某化工厂的好氧池中分离筛选得到2株苯胺降解菌TZ1和JH1,经16S rDNA测序鉴定为Comamonas sp.TZ1和Pseudomonas sp.JH1,均具有较强的苯胺降解能力,培养24 h后,可使初始浓度为800 mg/L的苯胺去除率达到96.4%~98.4%。在此基础上,按体积比1∶1将2株菌液进行混合构建了混合菌体系,进而对比考察了苯胺初始浓度、pH、盐度和重金属等环境因子对单一菌和混合菌生长量及降解苯胺效果的影响,重点探讨混合菌对不适宜生长环境的适应性及其对苯胺的降解特性。通过单一菌和混合菌对比实验发现,在适宜苯胺初始浓度、pH和盐度条件下,混合菌的生长量略高于单一菌;在不适宜生长的高浓度苯胺、pH和盐度条件下,混合菌也表现出了更强的适应性和苯胺矿化能力。Zn2+和Cr6+耐受实验则表明,对于Cr6+,混合菌表现出了更强的耐受能力,而对于Zn2+并没有表现出更强的耐受能力。     

液氨预处理强化复合菌系WSC-6降解稻秆的性能

高效、经济的预处理技术对提高稻秆的生物降解效率,实现稻秆资源的工业化利用具有重要意义。为确定液氨对稻秆生物降解性的影响,以复合菌系WSC-接种液氨预处理的稻秆,研究其生物降解性能。结果表明,用液氨预处理后的稻秆作为碳源,由于营养结构的改变,复合菌系WSC-6需要经过3d的适应期进入对数增长期,氮源利用率和微生物浓度均显著高于未处理的对照组。复合菌系WSC-6对其生物降解性得到明显提高,7d后的稻秆失重率达到62．7％,纤维素、半纤维素和木质素的降解率分别达到57．5％、85．6％和36．3％。液氨预处理对发酵系统的pH值影响不大,复合菌系WSC-对稻秆降解的pH值仍维持在6．5～8．6之间,有利于进行丁酸型发酵,这对后续利用乙酸高效生产沼气具有重要意义。     

Remazol Brilliant Blue R decolourization by the laccase from Trametes trogii

The decolourization of the recalcitrant dye RBBR by the culture filtrate of Trametes trogii and its isolated laccase was investigated. Both filtrates from Cu-induced cultures as well as purified laccase decolourized the dye RBBR. The purified laccase decolourized the dye down to 97% of 100 mg l(-1) initial concentration of RBBR when only 0.2U ml(-1) of laccase was used in the reaction mixture. The effects of different physicochemical parameters were tested and optimal decolourization rates occurred at pH 5 and at a temperature of 50 degrees C. Decolourization of RBBR occurred in the presence of metal ions which could be found in textile industry effluents. Of all the metal ions tested, FeCl2 was the most inhibiting for the decolourization. HBT was shown to have no effect on the decolourization of RBBR at low concentration, while at a concentration of 5 mM it slightly inhibited decolourization. The presence of aromatic compounds was found to be inhibiting for the decolourization at a concentration of 10 mM, but not at 0.1 mM, while at 1 mM only ortho-diphenols were inhibiting. Probing the effect of methanol it was found that higher concentrations caused a decrease in the decolourization rate of RBBR. The effect of laccase inhibitors on the decolourization of RBBR was tested with L-cysteine, SDS and EDTA. It was demonstrated that L-cysteine was the most inhibiting substrate for the decolourization while SDS was only inhibiting at 10 mM concentration and ETDA was not inhibiting at all tested concentrations.     

Biodegradation of hydrogen sulphide by inoculated Rhodococcus sp.zw11 in a pilot-scale biotrickling filter

A strain of autotrophic micro-organism, Rhodococcus sp.zw11, was isolated from pharmaceutical wastewater containing hydrogen sulphide (H₂S). The shape, physiological and biochemical characteristics and oxidation capacity of Rhodococcus sp.zw11 were studied, and the effect of inlet concentration and volumetric loading of H₂S on the removal efficiency was evaluated by the biotrickling filter inoculated with Rhodococcus sp.zw11. The results suggested that the optimal temperature of Rhodococcus sp.zw11 (aerobic bacilli, short rod and gram-negative) was from 20°C to 28°C and the optimal pH was from 5.5 to 6.5. The criteria necessary for a scale-up design of the biotrickling filter were established, and pressure drops at the start and end of the experiment were investigated. The optimal inlet loading could be noted as 180 g/m³h, corresponding to H₂S removal efficiency close to 100%. Furthermore, the inoculated biotrickling filter had good ability to resist shock loading, which was a potential industrialisation method to control H₂S emissions.     

新生MnO2对水中酸性媒介深黄GG的吸附脱色作用

以化学法合成的新生MnO2作吸附，对水中酸性媒介深黄GG进行吸附脱色研究，探讨了影响吸附的因素，结果表明，该吸附剂在PH1．5以下，投加量为0.3mg/L，温度为15℃条件下，饱和吸附量达1320mg/g，脱色率达96％以上，具有很高的吸附脱色能力。PH值是能力吸附能力的关键因素，温度，染料浓度和MnO2投加量影响程度较小。     

Biodegradation of s-triazine herbicide atrazine by Enterobacter cloacae and Burkholderia cepacia sp. from long-term treated sugarcane-cultivated soils in Kenya

In this study soils from sugarcane-cultivated fields were screened for bacterial species capable of atrazine (6-chloro-N²-ethyl-N⁴-isopropyl-1,3,5-triazine-2,4-diamine) degradation due to long exposure of the soils to this herbicide. To enrich for atrazine degraders, Minimal Salt Medium containing atrazine as the sole N source and glucose as the C source was inoculated with soils impacted with this herbicide and incubated. Bacterial growth was monitored by measuring optical density. The degradation of atrazine was followed by measuring residual atrazine in liquid cultures over a given time period by high performance liquid chromatography. Bacterial strains isolated from the enrichment cultures were characterized by biochemical tests and identified by 16S rRNA gene sequencing. Two bacterial strains coded ISL 8 and ISL 15 isolated from two different fields were shown to have 94 and 96% 16S rRNA gene sequence similarity to Burkholderia cepacia respectively. Another bacterial sp., ISL 14 was closely related to Enterobacter cloacae with a 96% 16S rRNA gene sequence similarity. There was not much difference between the extents of atrazine degradation by the enrichment cultures with communities (79–82% applied amount) from which pure strains were isolated and the pure strains themselves in liquid cultures that showed a degradation of 53–83% of applied amount. The study showed existence of bacterial strains in different sugarcane-cultivated fields which can use atrazine as a nitrogen source. The bacterial strains isolated can be used to enhance the degradation of atrazine in contaminated soils where atrazine is still considered to be recalcitrant.     

Biodegradation of s-triazine herbicide atrazine by Enterobacter cloacae and Burkholderia cepacia sp. from long-term treated sugarcane-cultivated soils in Kenya

In this study soils from sugarcane-cultivated fields were screened for bacterial species capable of atrazine (6-chloro-N2-ethyl-N?-isopropyl-1,3,5-triazine-2,4-diamine) degradation due to long exposure of the soils to this herbicide. To enrich for atrazine degraders, Minimal Salt Medium containing atrazine as the sole N source and glucose as the C source was inoculated with soils impacted with this herbicide and incubated. Bacterial growth was monitored by measuring optical density. The degradation of atrazine was followed by measuring residual atrazine in liquid cultures over a given time period by high performance liquid chromatography. Bacterial strains isolated from the enrichment cultures were characterized by biochemical tests and identified by 16S rRNA gene sequencing. Two bacterial strains coded ISL 8 and ISL 15 isolated from two different fields were shown to have 94 and 96% 16S rRNA gene sequence similarity to Burkholderia cepacia respectively. Another bacterial sp., ISL 14 was closely related to Enterobacter cloacae with a 96% 16S rRNA gene sequence similarity. There was not much difference between the extents of atrazine degradation by the enrichment cultures with communities (79-82% applied amount) from which pure strains were isolated and the pure strains themselves in liquid cultures that showed a degradation of 53-83% of applied amount. The study showed existence of bacterial strains in different sugarcane-cultivated fields which can use atrazine as a nitrogen source. The bacterial strains isolated can be used to enhance the degradation of atrazine in contaminated soils where atrazine is still considered to be recalcitrant.     

Biodegradation of crystal violet by a Shewanella sp. NTOU1

A bacterial isolate, strain NTOU1, originally isolated from the cooling system in an oil refinery could decolorize and detoxify crystal violet under anaerobic conditions. The strain was characterized and identified as a member of Shewanella decolorationis based on Gram staining, morphology characters, biochemical tests, the 16S rRNA gene and the gyrase subunit beta gene (gyrB). The optimum pH value and temperature for decolorization of crystal violet by this strain under anaerobic conditions were pH 8-9 and 30-40 degrees C, respectively. Formate (20 mM) was the best electron donor. Addition of ferric citrate did not inhibit decolorization of crystal violet, the addition of thiosulfate, ferric oxide, or manganese oxide slightly decreased decolorization, while addition of nitrite (20 mM) inhibited the decolorization of crystal violet. By supplementing the medium with formate and ferric citrate and cultivating it under optimum pH and temperature, this strain could remove crystal violet, at a concentration of 1500 mg l(-1), at the rate of 298 mg l(-1) h(-1) (during decolorization the OD(600) of the cell culture increased from approximately 0.6 to approximately 1.2). GC/MS analysis of the degradation products of crystal violet detected the presence of N,N'-bis(dimethylamino) benzophenone (Michler's Ketone), [N,N-dimethylaminophenyl] [N-methylaminophenyl] benzophenone, N,N-dimethylaminobenzaldehyde, N,N-dimethylaminophenol, and 4-methylaminophenol. These results suggest that crystal violet was biotransformed into N,N-dimethylaminophenol and Michler's Ketone prior to further degradation of these intermediates. This paper proposes a probable pathway for the degradation of crystal violet by this Shewanella sp. Cytotoxicity and antimicrobial tests showed that the process of decolorization also detoxify crystal violet.     

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.     

Acid and aluminum effects on the survival of littoral macro-invertebrates during acute bioassays

Six common macro-invertebrates were exposed to soft water at pH 4.5, with or without 200 microg liter(-1) Al added. Survivals were determined at 6, 12, 24 and 48 h and compared with neutral pH, Al-free controls. The order of acid-sensitivity among the test animals, from greatest to least (with mean 24/48 h survivals in the pH 4.5, low Al treatment in parentheses), was: Caenis sp. (2%) > Hyalella azteca (12%) > Enallagma sp. (20%) > Gyraulus sp. (55%) > Chironomidae (94%) > Hydracarina (99%). Aluminum significantly reduced the survivals of Gyraulus, Hyalella and Chironomidae. The latter group experienced no significant mortality at pH 4.5 except when Al was present. In contrast, the Hydracarina were unaffected by both acid and acid plus Al exposure, and the survivals of Enallagma and Caenis at low pH were enhanced by Al. These differential responses to the treatments indicate that both acid and Al stress may control the structure of the littoral macroinvertebrate community in acid lakes.     

造纸黑液酸析法回收木质素

研究了加酸沉淀回收碱法草浆黑液中木质素的工艺,探讨了酸浓度,温度,ｐＨ值及加酸速度对木质素回收量的影响,得出的最佳条件是：硫酸浓度为５０％,温度为４０℃￣５０℃,ｐＨ值为３．０左右,加酸速度以控制不使产生大量泡沫为准。在上述条件下,木质素的回收效果最佳。