活性污泥体系中聚糖菌的富集与鉴定

活性污泥体系中,聚糖菌(GAOs)在厌氧环境下与聚磷菌(PAOs)形成对底物的竞争关系,对聚糖菌的研究对于优化生物除磷工艺有重要意义。以葡萄糖为惟一碳源,在磷限制条件下,利用特殊运行方式对活性污泥进行驯化培养出了稳定的聚糖菌颗粒污泥,厌氧阶段磷释放量与有机物吸收量浓度(mg/L)比从7.4%下降为0.25%。从培养好的活性污泥反应器中分离获得2株聚糖菌,经菌落PCR和16S rRNA序列分析确定了所得聚糖菌菌株G1和菌株G2分别是枯草芽孢杆菌(Bacillus subtilis)和解鸟氨酸克雷伯氏菌(Klebsiella ornithinolytica)。     

一株高效脱硫脱氨氮菌的分离、鉴定及系统发育分析

从运行稳定的能同步脱臭的曝气生物滤池中采集样品,富集分离获得一株能高效脱硫脱氨氮的菌株TS-1。对分离菌株进行形态观察、生理生化试验及16S rRNA基因序列分析,结果表明,该菌株TS-1为革兰氏阳性菌,杆状;菌落在营养琼脂培养基上呈圆形,表面光滑,乳白色半透明;V-P试验阴性,能水解淀粉和明胶,利用柠檬酸盐生长;对菌株进行16S rRNA的PCR扩增,扩增产物测序结果表明分离菌株TS-1与巨大芽孢杆菌（Bacillus megaterium）同源性达到99%;以16S rRNA同源性为基础构建了包括24株相关种属细菌在内的系统发育树,在系统发育树中,分离菌株TS1与Bacillus megaterium在同一分支。结合形态观察、生理生化试验及16S rRNA基因序列分析结果,将其初步鉴定为巨大芽孢杆菌（Bacillus megaterium）。在常温（30±2）℃、转速为150 r/min的条件下,处理pH 7、S^2-和NH⁺₄-N分别为80 mg/L和88 mg/L的水样40 h,硫化物和氨氮的脱除率分别为91.8%和96.6%。     

对氨基苯磺酸降解菌的分离及其特性研究

从温州地区受污染的河水中分离到一株能降解对氨基苯磺酸的菌株WZR-3,该菌株能以对氨基苯磺酸为惟一碳源、能源生长。经对其形态特征、生理生化以及16S rDNA序列分析,该菌株初步鉴定为人苍白杆菌(Ochrobactrum anthropi)。该菌株利用对氨基苯磺酸生长时最适生长温度和pH值分别为30℃和7。该菌在10 g/L对氨基苯磺酸时仍能生长,最适生长浓度为300 mg/L对氨基苯磺酸。降解底物广谱性测试表明,该菌株还能降解多种芳香类化合物。     

一株异养脱氮菌的脱氮性能及其影响因素研究

研究异养硝化过程中分离得到一株具有异养脱氮性能的菌株,经16S rRNA鉴定后为Providencia rettgeri菌属,命名为Providencia rettgeri strain YL。在实验室条件下,初步探讨了不同碳氮源、温度、pH值、摇床转速和微量元素对Providencia rettgeri strain YL菌株脱氮作用的影响,研究结果显示,Providencia rettgeri strain YL菌株在葡萄糖和氯化铵为碳氮源的培养基中生长最好,温度为30℃、pH值为7、摇床转速为120 r/min时具有最佳的脱氮效果,微量元素Mg²⁺最有利于Providencia rettgeri strain YL菌株的生长和脱氮。     

一株反硝化细菌的分离鉴定及其反硝化特性简

从处理城镇污水的移动床生物膜反应器中分离获得一株反硝化细菌D₃,并进一步研究该菌株的系统发育地位及反硝化特性。采用16S rDNA序列分析对菌株进行初步鉴定,探讨了基质浓度、起始pH和温度对菌株反硝化活性的影响。根据形态学特征、生理生化特性及16S rDNA序列测定分析,初步鉴定菌株属于寡养单胞菌属。该菌能利用硝酸钠或亚硝酸钠进行反硝化作用,最佳电子受体是硝酸盐氮,反硝化速率最大为19.86 mg/（L·h）,最适生长pH为7.36,最适生长温度为33.5℃。菌株D₃在初始硝态氮浓度为140 mg/L,以乙酸钠为惟一碳源,pH为7.36,温度33.5℃的最优生长条件下,培养10 h进入对数生长期,倍增时间为9.9 h,48 h内硝酸盐还原率达95%。     

畜禽养殖污水中高效氨氮降解菌的筛选、鉴定及生长条件研究

从养殖污水中分离纯化得到1株高效氨氮降解菌,对其进行形态特征、生理生化、16S rDNA序列分析以及最佳生长条件研究,并将菌株投入养殖污水降解污水中的氨氮。结果表明：AN4菌株在NH4＋-N初始质量浓度为50 mg/L的条件下,24 h内的氨氮降解率为92.5%;初步鉴定该菌株为苍白杆菌属（Ochrobactrum sp.）,菌株的16S rDNA序列在Gen-Bank的登录号为GU345782;AN4菌株在装液量为60.64 mL,pH为7.06,葡萄糖为6.0 g/L的条件下培养,菌株的降解率可以达到94.28%;菌株对养殖污水中氨氮的降解率为12.3%。苍白杆菌菌属能够降畜禽养殖污水中的氨氮还未见报道,AN4菌株的筛选获得为生物降解养殖污水中的氨氮又提供了一种新型菌株。     

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。     

自养反硝化菌对硝酸盐氮去除动力学及影响因素研究

为更经济有效地去除污水中的硝酸盐,从兼性污泥中分离获得6株能氧化单质硫和还原硝酸盐的自养反硝化菌。根据各菌株的降解曲线筛选出优势菌种N-I,并研究影响菌株N-I降解性能的环境因素,如pH、温度、碳源及硝酸盐的降解动力学。实验表明,菌株N-I对硝酸盐的降解符合一级反应动力学方程,反应的半衰期t_1/2为1.42 h,反应速率常数为0.488 h^-1。最佳反应pH=7,最佳反应温度为30℃,最佳NaHCO₃浓度为大于或等于2.5 g/L。     

一株高效脱硫菌的分离鉴定和脱硫特性研究

从污水净化厂二沉池回流污泥中分离到一株硫杆菌菌株dj-5,该菌株是一种兼性厌氧菌,通过生理生化特性鉴定,并结合16S rDNA序列分析及鉴定,该菌株可以确定为脱氮硫杆菌（Thiobacillus denitrificans）。该菌株的生长曲线表明菌体生长迅速,延滞期约为8 h,然后细菌进入对数生长期,这一阶段持续时间大约20～24 h,稳定期较短,细菌很快进入衰亡期。通过脱硫工艺实验考察了pH值、温度和搅拌速度对脱硫率和菌体生长的影响,结果表明,在进气量180 L/h、pH=6～8、温度为25～35℃和搅拌速度为270 r/min时,该菌株能正常生长,对进气浓度高达2 500 mg/m³的硫化氢脱除率在91%以上。     

活性污泥中高效除臭菌株的驯化和筛选

以复合生物滤池作为生物除臭反应器,活性污泥作为菌源,进行氧化S^2-和NH⁺₄的活性驯化,2周后驯化成功,去除率达80%以上。通过平板划线分离,共分离出5株菌,其中细菌、酵母菌平板上各2株,霉菌平板上1株。将分离出的5株菌分别接种至含S^2-、含NO^-₂和含NH⁺₄的3类液体培养基中进行摇床培养2周,通过对菌株氧化能力的实验研究,筛选出2株净化H₂S的高效菌株,2株净化NH₃的高效菌株,能利用恶臭气体作为基质,具有潜在的应用价值。     

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。     

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一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CMl,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16SrDNA序列分析鉴定该菌株属于耐硼赖氨酸芽孢杆菌（Lysinibacillusb oronitolerans）。通过研究温度和pH值对菌株CMl降解MC—RR能力的影响,发现菌株CMl在60h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。CMl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,12h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。     

Bacterial diversity in paclobutrazol applied agricultural soils

The aim of this study was to investigate the bacterial communities on paclobutrazol [(2RS, 3RS)-1-(4-Chlorophenyl)-4, 4-dimethyl-2-(1H-1,2,4-triazol-1-yl) pentan-3-ol]–applied agricultural soils by denaturing gradient gel electrophoresis (DGGE) of polymerase chain reaction (PCR) amplified 16S rDNA gene fragments. Three different agricultural soil samples were collected from paclobutrazol applied mango and waxapple orchards, peanut fields and untreated rice fields as a control for DGGE analysis. The DGGE pattern of PCR- generated 16S rDNA gene fragments indicated that the bacterial populations from four paclobutrazol–applied soils of peanut fields were closely related to each other and two paclobutrazol–applied soils of mango and waxapple orchards harbored closely related bacterial communities. But, paclobutrazol–free agricultural soils comprised relatively a different bacterial group. However, the bacterial populations of mango and waxapple orchard are completely different from the bacterial communities of peanut field. Further purification and sequence analysis of 40 DGGE bands followed by phylogenetic tree assay showed similar results that soil bacteria from paclobutrazol applied mango and waxapple orchard are phylogenetically related. Based on the phylogenetic analysis, the clone M-4 was clad 100 % (bootstrap value) with Mycobacterium sp. The Mycobacterium sp. has been proved to degrade the phenolic compounds such as phenol, 4-chlorphenol, 2,4-dichlorophenol and paclobutrazol molecule containing chlorobenzene ring.     

高温好氧反硝化菌的分离鉴定及脱氮特性

从太原市某污水处理厂SBR活性污泥中分离纯化得到一株高温(50℃)好氧反硝化菌,命名为XF3。通过生理生化特性鉴定及16S rDNA序列分析,初步鉴定为波茨坦短芽孢杆菌。通过单因子实验考察碳源、C/N、pH及接种量对该菌株的生长情况与反硝化性能的影响。结果表明,菌株XF3最适碳源为琥珀酸钠,最佳C/N为12∶1,最佳pH为7,最适接种量10%(体积分数)。同时该菌株具有良好的异养硝化能力,48 h可以将73 mg/L氨氮几乎全部降解。     

红三叶草根际区石油降解菌的筛选及降解性能

从石油污染的土壤红三叶草（nifoliumrepensLinn）根际修复区中分离筛选得到4株以原油作为惟一碳源和能源进行生长繁殖的高效石油降解菌。通过菌落形态、显微镜个体形态观察、生理生化鉴定以及菌株16SrDNA序列分析,初步鉴定4株优势降解菌分别为动性杆菌、藤黄微球菌、蜡状芽孢杆菌和短小芽孢杆菌。采用气相色谱／质谱（GC／MS）法分析4株混合菌对石油烃的降解性能。结果表明：在摇床培养条件下,混合菌54d对总石油烃的生物降解率达到90．50％,较对照高67．72％。随着生物降解时间的延长,石油组分中的正构烷烃、异构烷烃及环烷烃相对总量均呈减小趋势,而芳香烃和其他醇类、醛和酸类的相对含量则有所增加。     

一种未见报道的厌氧氨氧化菌分子生物学鉴定

以(NH4):SO4和NaNO2作为基质,富集厌氧氨氧化污泥.提取厌氧氨氧化污泥中细菌总DNA,纯化后使用特异性引物对厌氧氨氧化菌16S rDNA进行PCR扩增.扩增产物连接到pMD19-T载体,将载体转化到感受态细胞大肠杆菌JM109中,并对其16S rDNA基因进行测序.将测序结果进行系统发育树分析,发现富集得到的厌氧氨氧化菌与Candidatus Anammoxoglobus propionicus进化关系比较接近,是一种尚未见报道的厌氧氨氧化菌.     

Isolation and characterization of a newly isolated pyrene-degrading Acinetobacter strain USTB-X

The pryene-degradation bacterium strain USTB-X was newly isolated from the polycyclic aromatic hydrocarbon (PAH)-contaminated soil in Beijing Coking Plant, China. The strain was identified as Acinetobacter with respect to its 16S rDNA and morphological and physiological characteristics. The strain was Gram-negative, non-mobile, non-acid-fast, and non-spore-forming, short rods in young culture and 0.8–1.6 μm in diameter and 1.2–2.5 μm long in the stationary phase of growth. Strain USTB-X could utilize pyrene, naphthalene, fluorene, phenanthrene, benzene, toluene, ethylbenzene, ethanol, methanol, and Tween 80 as sole source of carbon and energy. The strain could produce biosurfactants which enhanced the removal of pyrene and could remove 63 % of pyrene with an initial concentration of 100 mg·L^?1 in 16 days without other substrates. Based on the intermediates analyzed by gas chromatography-mass spectrometry, we also deduced the possible metabolic pathway of strain USTB-X for pyrene biodegradation. Results indicated that the strain USTB-X had high potential to enhance the removal of PAHs in contaminated sites.     

有机碳源条件下厌氧氨氧化SBBR中的微生物多样性

研究有机碳源对SBBR厌氧氨氧化菌群等微生物的影响。采用16S rDNA序列与PCR-DGGE分析技术相结合的方法,对稳定运行的反应器内的活性污泥和生物膜样品,进行细菌多样性图谱分析,同时采用巢式PCR-DGGE技术对浮霉状菌属(Planctomycetes)细菌进行分析。结果表明,在有机碳源反应系统细菌条带数和多样性指数均高于无机系统,与活性污泥相比,生物膜表尤为明显。当进水不含有机碳源时,氨氧化细菌(ammonia oxidizing bacteria,AOB),厌氧氨氧化菌(anaerobic ammonia oxidizing bacteria,ANAMMOX)为优势功能菌;当进水含有机碳源时,系统中存在的AOB以亚硝化单胞菌(Nitrosomonas sp.)为优势菌群,同时存在反硝化菌,如索氏菌(Thauera sp.)以及厌氧氨氧化菌,它们共同作用完成N的去除。此外,与无机碳源系统相比,有机碳源的存在,有利于浮霉状菌的积累,但压缩了ANAMMOX的生存空间。本研究可为厌氧氨氧化工艺处理低C/N比有机废水提供了理论依据。     

Biodegradation of nicotine by a novel Strain Shinella sp. HZN1 isolated from activated sludge

The nicotine-degrading bacterium HZN1 was isolated from activated sludge and identified as Shinella sp. based on its physiological characteristics and analysis of 16S rDNA gene. Strain HZN1 is capable of using nicotine as the sole carbon source in the mineral salts medium. The optimum temperature and pH for strain HZN1 growth and nicotine degradation were 30°C and 7.0, respectively. It could degrade approximately 100 % of 0.5 g L^?1 of nicotine within 9 h. Three intermediate metabolites were produced by the strain HZN1 and identified as cotinine, myosmine and nicotyrine using gas chromatography-mass spectrometry. This is the first report of nicotine-degrading strain from the genus of Shinella. The results showed that strain HZN1 could be potentially employed in bioremediation of nicotine. Our findings would provide a new insight into the biodegradation of nicotine.     

Isolation and characterization of lead-tolerant Ochrobactrum intermedium and its role in enhancing lead accumulation by Eucalyptus camaldulensis

In this study, the potential of rhizospheric bacteria in promoting the growth and Pb accumulation by the woody plant Eucalyptus camaldulensis under hydroponic conditions was investigated for the first time. Three Pb-tolerant bacteria were isolated from the rhizosphere of E. camaldulensis grown in Pb-contaminated soils in the Bo Ngam Pb mine, Thailand. Based on analysis of partial 16S rRNA gene sequence, the three isolates were identified as Microbacterium paraoxydans BN-2, Ochrobactrum intermedium BN-3, and Bacillus fusiformis BN-4. Among these strains, O. intermedium BN-3 showed the highest tolerance to not only Pb but also Cd and Zn. After growth in the presence of Pb, the membranes of O. intermedium BN-3 cells exhibited an increase in unsaturated fatty acid levels but a decrease in fluidity. In hydroponic studies, inoculation of O. intermedium BN-3 significantly increased the biomass and Pb accumulation by E. camaldulensis compared to the uninoculated control. The results suggested the role of the natural rhizospheric bacteria localized to the root surface of E. camaldulensis in promoting Pb accumulation and plant growth. Our results indicate that O. intermedium BN-3 and other indigenous rhizospheric bacteria have the potential to improve the efficiency of phytoremediation of Pb-contaminated sites.