一株高效降酚菌的分离鉴定及降酚特性研究

以苯酚为唯一碳源,采用逐量分批的方法从被酚类物质污染的土壤中驯化、筛选、分离降酚菌,利用形态学、培养特性、生理生化反应以及分子手段鉴定所得菌株,研究了该菌株生长与降酚的关系以及其降酚能力,并设计引物检测降酚酶基因。结果表明,获得的一株高效降酚菌GDYW-0027经鉴定为不动杆菌属(Acinetobacter),它的生长速率与苯酚的降解速率基本同步,48h内对高质量浓度(2.2g/L)苯酚的降解率为86.73%;在菌株GDYW-0027中检测到苯酚羟化酶及邻苯二酚2,3双加氧酶基因片段,推测其采用间位开环方式打开苯环。     

降氰菌的分离及培养条件初探

从温州某电镀厂的活性污泥中分离纯化、筛选获得降氰菌16株,其中11菌株能耐受2400mg/L的氰化钠.同时,考察了实验条件对11菌株的生长影响,结果表明,11菌株最佳生长pH为8.0,4～18h为对数生长期;果糖对11菌株的生长和降氰有促进作用;50mL含氰液体培养基中,1mol/L的Fe或Cu对11菌株的生长和降氰均有抑制作用;11菌株的最佳降氰液体培养基pH为6.5,4h内的降氰率可达98.57%;8h后CN-残留质量浓度降至501μg/L.     

高效联苯降解菌的筛选及其特性

焦化废水属高浓度难降解工业有机废水,其中所含的多环芳烃属难降解有机物,且对环境产生毒害作用。为探索生物强化技术去除焦化废水中多环芳烃类化合物,采用选择性培养和多级富集的方法,以联苯为模型化合物,并作为惟一碳源,从焦化厂废水和污泥中分离和筛选得到6株联苯降解菌。通过逐渐提高底物浓度的方法驯化菌株后,从中筛选出降解效率最高的联苯降解菌株WIS-01,在此基础上进行菌属鉴定、细菌生长情况及联苯降解性能的研究。实验结果表明,3 d内菌株WIS-01对联苯的降解率可达99%以上,可耐受联苯的最高质量浓度为2 g/L。通过形态学、生理生化鉴定和16S rDNA序列比对分析,确定菌株WIS-01属于假单胞菌属,同源性达95%,命名为Pseudomonas sp.WIS-01。     

聚合物驱采出水中聚丙烯酰胺的微生物联合降解作用研究

通过对2株细菌的培养降解实验研究聚丙烯酰胺(hydrolyzed polyacrylamide,HPAM)降解菌对水环境下聚丙烯酰胺的降解作用,讨论协同降解机理。2株降解聚丙烯酰胺的菌株假单胞菌CJ419、枯草芽孢杆菌FA16在初始30℃废水样品上培养,定期测量细菌生物量和HPAM降解率。培养30 d后CJ419和FA16对聚合物的降解率最大值分别达到30.4%和25%,而以1∶1比例的混合菌降解率最大值达到80.3%。对2株菌胞外各组分研究表明:混合菌降解HPAM的机理主要由胞外降解酶系水解聚合物侧链基团导致HPAM降解为小分子物质,同时生长过程中降解菌还会释放非蛋白还原性物质引发氧化反应共同参与HPAM降解。     

一株高效MC-RR降解菌的分离鉴定及其降解特性

为了得到一株具有降解微囊藻毒素-RR(MC-RR)特性的产芽孢菌株,采用加热富集芽孢菌的方法,从太湖分离到一株MC-RR降解菌CM1,该菌对MC-RR具有强烈的降解特性。通过形态学特征、生理生化特征及16S rDNA序列分析鉴定该菌株属于耐硼赖氨酸芽孢杆菌(Lysinibacillus boronitolerans)。通过研究温度和pH值对菌株CM1降解MC-RR能力的影响,发现菌株CM1在60 h将MC-RR由12.77μg/mL降解到1.67μg/mL,降解率达86.90%,最适降解温度为37℃,最适pH值为7.0。CM1菌株的胞外物质和胞内物质均能降解MC-RR,但胞内物质具有更强烈的降解特性,12 h可以将7.27μg/mL的MC-RR完全降解。为丰富MC-RR降解菌纯菌种研究以及在去除水体中MC-RR应用研究方面提供了理论基础。     

降酚菌株的固定化细胞处理含酚废水的性能研究

用海藻酸钠作为载体将一株降酚菌株进行固定化包埋。利用正交实验确定了该菌株固定化细胞制备的最优条件。研究表明,该降酚菌株的固定化细胞对苯酚的降解能力和耐受能力均大于游离细胞。降酚菌株固定化细胞降解苯酚的最适温度范围是30~35℃,最适pH值范围为6~8。该菌株的固定化细胞对废水中的COD也具有良好的降解效果。     

利用Rank氧电极筛选焦化废水功能菌的探索

介绍了一种利用Rank氧电极筛选焦化废水功能菌的方法。实验所用焦化废水中含有结构不明的成色有机物，从驯化后的活性污泥中分离到一组对该物质有效的菌株，利用Rank氧电极测定其对该物质的活性大小，筛选出活性较好的菌种用于废水生化处理的进一步研究。对于结构不明的物质降解，这种方法具有快速、简捷、灵敏度高等优点。可广泛应用于功能菌的筛选等方面。     

降酚菌株的固定化细胞处理含酚废水的性能研究

用海藻酸钠作为载体将一株降酚菌株进行固定化包埋.利用正交实验确定了该菌株固定化细胞制备的最优条件.研究表明,该降酚菌株的固定化细胞对苯酚的降解能力和耐受能力均大于游离细胞.降酚菌株固定化细胞降解苯酚的最适温度范围是30～35℃,最适pH值范围为6～8.该菌株的固定化细胞对废水中的COD也具有良好的降解效果.     

油田污染土壤中石油烃类微生物及其降解基因的分布

以原油为唯一碳源,运用传统分离培养法,从中国江苏省淮安市金南油田石油污染土壤中筛选出9株具有原油降解功能的菌株并分别命名为JN1~JN9。对其进行形态观察、生理生化实验和分子生物学鉴定,最终确定其分别隶属于Lysinibacillus、Pseudomonas、Raoultella等5个菌属;在适宜条件下进行降解实验,9株菌对原油的降解率最高可达到32.63%;进一步对9株菌进行质粒和基因组上原油降解基因的分析,结果表明,GSTs基因广泛分布在其中8株菌的基因组上,同时JN1、JN3、JN4的质粒上也检测到了GSTs基因;alk B基因分别在7株菌的基因组中和2株菌的质粒检出;Lm PH基因仅在JN9的基因组上检出。利用分子生物学手段对石油降解菌降解基因的分布的初步研究结果,可为原油污染场地的微生物修复提供理论依据。     

高效降氰菌群的构建及降解特性

从筛选到的降氰菌中构建出了优于单一菌种降氰活性的复配菌群CNR.研究了该复合菌群的生长条件,探讨了温度、pH、接种量、氰化物初始浓度及降解时间等因素对CNR降氰的影响.研究表明,复配菌群CNR适应碱性环境,可降解高浓度氰化物(CN-11 000 mg/L),并对金属氰化物和脂肪族腈具有极强的降解能力.在有氧、pH 11、33 ℃和接种量10%条件下.含CN-11 000 ng/L培养液经60 h降解后,CN-1浓度降为0.49 mg/L,降氰率高达99.96%,达到国家一级排放标准.     

应用络合沉淀-化学氧化组合工艺处理高浓度含氰农药废水

由于氰化物的剧毒性以及农药废水的难生物降解性，含氰农药废水的治理难度较大。从工程应用的角度出发，在小试实验研究的基础上，确定了处理污染物含量高、难生物降解含氰农药废水的组合工艺流程为：FeSO₄络合沉淀→H₂O₂催化氧化→ClO₂深度氧化。组合工艺流程处理效果好，操作方便，出水水质稳定。研究结果表明，当FeSO₄加入量为理论计算值的1.3倍、3％H₂O₂和ClO₂投加量分别为24 mL/L和200 mg/L时，废水中CN^-离子的总去除率高达99.99％以上，COD总去除率达到99%。组合工艺不仅适用于处理高浓度含氰农药废水，而且也能为高浓度、难降解有机废水的治理提供有益的参考。     

Application of Klebsiella oxytoca immobilized cells on the treatment of cyanide wastewater

Klebsiella oxytoca, isolated from cyanide-containing industrial wastewater, has been shown to be able to biodegrade cyanide to non-toxic end products. The technology of immobilized cells can be applied in biological treatment to enhance the efficiency and effectiveness of biodegradation. In this study, potassium cyanide was used as the target compound and both alginate and cellulose triacetate techniques were applied for the preparation of immobilized cells. Results from this study show that KCN can be utilized as the sole nitrogen source by K. oxytoca. The free suspension systems reveal that the cell viability was highly affected by initial KCN concentration and pH. Results show that immobilized cell systems could tolerate a higher level of KCN concentration and wider ranges of pH. In the batch experiments, the maximum KCN removal efficiencies using alginate and cellulose triacetate immobilized beads were 0.108 and 0.101mM h(-1) at pH 7, respectively. Results also indicate that immobilized system can support a higher biomass concentration. Complete KCN degradation was observed after the operation of four consecutive degradation experiments with the same batch of immobilized cells. This suggests that the activity of immobilized cells can be maintained and KCN can be used as the nitrogen source throughout KCN degradation experiments. The maximum KCN removal rates using alginate and cellulose triacetate immobilized beads in continuous-column system were 0.224 and 0.192mMh(-1) with initial KCN concentration of 3mM, respectively. Results indicate that the immobilized cells of K. oxytoca would be applicable to the treatment of cyanide-containing wastewaters.     

好氧活性污泥中自养硝化细菌分离方法研究

分别采用琼脂平板、琼脂糖平板、水洗洋菜平板和硅胶平板分离培养城市污水处理厂好氧活性污泥中的硝化细菌。在相同的培养条件下,琼脂平板和水洗洋菜平板有杂菌生长;硅胶平板制作过程繁琐,培养后期出现失水性干裂;琼脂糖平板没有杂菌生长。通过对分离菌株的表观形态观察和NorB基因检测鉴定分离菌株,证明琼脂糖分离出了目的菌株,适合生长周期长的自养硝化细菌分离培养。     

Aquatic environments polluted with antibiotics and heavy metals: a human health hazard

Aquatic environments often receive wastewater containing pollutants such as antibiotics and heavy metals from hospital sewage, as well as contaminants from soil. The presence of these pollutants can increase the rate of exchange of resistant genes between environmental and pathogenic bacteria, which can make the treatment of various types of bacterial infections in humans and animals difficult, in addition to causing environmental problems such as ecological risk. In this study, two tetracycline-resistant Pseudomonas aeruginosa (EW32 and EW33), isolated from aquatic environments close to industries and a hospital in southeastern Brazil, were investigated regarding the possible association between tetracycline and heavy metal resistance. The isolate EW32 presented a conjugative plasmid with coresistance to tetracycline and copper, reinforcing the concern that antibiotic resistance by acquisition of plasmids can be induced by the selective pressure of heavy metals in the environment.     

TOL plasmid transfer during bacterial conjugation in vitro and rhizoremediation of oil compounds in vivo

Molecular profiling methods for horizontal transfer of aromatics-degrading plasmids were developed and applied during rhizoremediation in vivo and conjugations in vitro. pWW0 was conjugated from Pseudomonas to Rhizobium. The xylE gene was detected both in Rhizobium galegae bv. officinalis and bv. orientalis, but it was neither stably maintained in orientalis nor functional in officinalis. TOL plasmids were a major group of catabolic plasmids among the bacterial strains isolated from the oil-contaminated rhizosphere of Galega orientalis. A new finding was that some Pseudomonas migulae and Pseudomonas oryzihabitans strains harbored a TOL plasmid with both pWW0- and pDK1-type xylE gene. P. oryzihabitans 29 had received the archetypal TOL plasmid pWW0 from Pseudomonas putida PaW85. As an application for environmental biotechnology, the biodegradation potential of oil-polluted soil and the success of bioremediation could be estimated by monitoring changes not only in the type and amount but also in transfer of degradation plasmids.     

Molecular characterization of soil bacteria antagonistic to Rhizoctonia solani,sheath blight of rice

Bacillus pumillus MTCC7615 has been identified as a potent isolate against Rhizocotonia solani, the fungal pathogen causing sheath blight in rice. The study aimed at probing the role of a 23kb size plasmid pJCP07 of Bacillus pumillus MTCC7615 in its fungal antagonism towards Rhizocotonia solani. Plasmid pJCP07 was found to be involved in production of a fungal antagonistic compound as demonstrated by plasmid curing and conjugational transfer experiments. Tn5 insertional studies further confirmed that the plasmid pJCP07 of Bacillus pumillus MTCC7615 carries some of the gene(s) involved in production of compound antagonistic to Rhizocotonia solani. The plasmid pJCP07 is thus a mobilizable medium-sized plasmid carrying genes responsible for antagonism of Bacillus pumillus MTCC7615 towards Rhizocotonia solani.     

Effect of some technical and formulated insecticides on microbial activities in soil

Abstract

The influence of 11 formulated and technical insecticides at 10 μg/g soil on growth and activities of microorganisms was determined. The populations of bacteria and fungi initially decreased with some pesticide treatments but recovered rapidly to levels similar to or greater than those of controls after three weeks. Both formulated and technical chlordane, chlorpyrifos and cypermethrin stimulated fungal growth. No inhibition on nitrification after two wks and sulfur oxidation after three wks was observed in treatments with either grade of insecticide. The effect of different treatments on respiration was equal to or greater than that of control sample. Less effect was observed with technical insecticides than the formulated ones on microbial populations and activities in the soil.     

The effect of polluted and leached snow melt waters on the soil bacterial community-quantitative response

The effect of polluted snow melt waters on the number of soil bacteria was determined using soil cores extracted from an upland catchment in the Cairngorm Mountains, Scotland. Total numbers of viable heterotrophic bacteria and bacterial denitrifiers were determined using plate and MPN counts. Separate soil cores were treated with simulated melt waters representative of either the composition of the first melt fraction from polluted or leached snowpacks. The number of bacteria in the Ah soil horizon (Hodgson, 1974) treated with polluted snow melt (PSM) water decreased significantly by 28-fold, but increased by 11-fold in the BC horizon. Denitrifier numbers decreased by 8-fold in the Ah horizon, but increased by over 2-fold lower down the profile. Overall the bacterial community exposed to simulated leached snow melt (LSM) waters showed little change in the Ah horizon. In the BC horizon (Hodgson, 1974), total viable bacterial numbers decreased by 20-fold, but denitrifiers numbers were unaffected.     

Comparisons of the dust/smoke particulate that settled inside the surrounding buildings and outside on the streets of southern New York City after the collapse of the World Trade Center, September 11, 2001

The collapse of the World Trade Center (WTC) on September 11, 2001, generated large amounts of dust and smoke that settled in the surrounding indoor and outdoor environments in southern Manhattan. Sixteen dust samples were collected from undisturbed locations inside two uncleaned buildings that were adjacent to Ground Zero. These samples were analyzed for morphology, metals, and organic compounds, and the results were compared with the previously reported outdoor WTC dust/smoke results. We also analyzed seven additional dust samples provided by residents in the local neighborhoods. The morphologic analyses showed that the indoor WTC dust/smoke samples were similar to the outdoor WTC dust/smoke samples in composition and characteristics but with more than 50% mass in the <53-microm size fraction. This was in contrast to the outdoor samples that contained >50% of mass above >53 microm. Elemental analyses also showed the similarities, but at lower concentrations. Organic compounds present in the outdoor samples were also detected in the indoor samples. Conversely, the resident-provided convenience dust samples were different from either the WTC indoor or outdoor samples in composition and pH, indicating that they were not WTC-affected locations. In summary, the indoor dust/smoke was similar in concentration to the outdoor dust/smoke but had a greater percentage of mass <53 microm in diameter.     

Performance of different microalgal species in removing nickel and zinc from industrial wastewater

A series of batch experiments was conducted to compare the ability of 11 microalgal species of the same cell density in removing nickel (Ni) and zinc (Zn) from synthetic wastewater. These included Chlorella vulgaris (commercially available), Chlorella sorokiniana and Scenedesmus quadricauda (isolates from polluted water of Wuhan, China), and eight different isolates from Hong Kong. The Wuhan isolate of Scenedesmus removed most Ni, probably due to its large biomass. Nickel concentration was reduced from an initial 30 to 0.9 mg/l after 5 min (97% Ni removal), and further declined to 0.4 mg/l after 90 min of treatment. In wastewater containing 30 mg/l Ni and 30 mg/l Zn, more than 98%, Ni and Zn were removed simultaneously at the end of 5 min treatment, indicating that the presence of Zn in wastewater did not affect Ni removal by this Scenedesmus isolate. The second most effective species for Ni removal was an isolate, tentatively identified as Chlorella miniata, Ni concentration was reduced to 10 mg/l after 90 min, and was only slightly interfered by the presence of Zn. In terms of metal removal per unit biomass or unit surface area of algal cells, C. miniata was the best species in removing Ni and Zn. At the other extreme, one Hong Kong isolate (Synechocystis sp.) did not remove any Ni and only achieved 40% Zn removal. Performance of the other isolates was comparable with the commercial C. vulgaris, less than 50% Ni was removed after 5 h of treatment and Ni removal was significantly reduced by the presence of Zn. All algae tested were found to be viable, showing these 11 species could tolerate a mixture of 30 mg/l Ni and 30 mg/l Zn in wastewater.