深层灰岩裂隙含水层的微生物种群特征及多样性分析

淄博深层灰岩裂隙含水层遭受石油污染已有30年,含水层中微生物降解对污染物的去除起到重要的作用,目前有关深层灰岩裂隙含水层中微生物群落结构和多样性的相关研究较少。本文采用高通量测序技术对研究区灰岩岩溶裂隙介质表面及地下水中的微生物种群结构进行了研究,对石油降解菌的种群特征及多样性进行了探讨,并利用RDA对典型石油降解菌与理化因子的相关性进行了分析。结果表明,含水层中的菌群主要属于变形菌门(Proteobacteria)和子囊菌门(Ascomycota),且地下水比灰岩岩溶裂隙介质表面呈现出更高的生物多样性;含水层中石油降解细菌主要属于脱硫弧菌属(Desulfovibrio)、类诺卡氏菌属(Nocardioides)、假黄色单胞菌属(Pseudoxanthomonas)等,石油降解真菌主要来自煤炱目(Capnodiales)、座囊菌目(Dothideales)、银耳目(Tremellales)中的菌种;地下水中的石油降解细菌、真菌与总石油烃(TPH)、全磷(TP)和全氮(TN)的相关性比灰岩岩溶裂隙介质更高。     

Assessing the bioremediation potential of arsenic tolerant bacterial strains in rice rhizosphere interface

The arsenic tolerant bacterial strains Staphylococcus arlettae(NBRIEAG-6), Staphylococcus sp.(NBRIEAG-8) and Brevibacillus sp.(NBRIEAG-9) were tested for their roles in enhancing plant growth and induction of stress-related enzymes in rice(Oryza sativa L. cv. NDR-359) plants at two different concentrations, 30 and 15 mg/kg of As(V) and As(III), respectively. An experiment was conducted to test the effect of these strains on plant growth promotion and arsenic uptake. We found 30%–40% reduction in total As uptake in bacteria-inoculated plants, with increased plant growth parameters compared to non-inoculated plants. Moreover, the bacteria-inoculated plants showed reduced activity of total glutathione(GSH) and glutathione reductase(GR) compared to their respective controls, which suggests the bacteria-mediated reduction of oxidative stress in plants. Thus, these strains were found to be beneficial in terms of the biochemical and physiological status of the plants under arsenic stress conditions.Furthermore, one-way ANOVA and principal component analysis(PCA) on enzymatic and non-enzymatic assays also revealed clear variations. The results support the distinction between control and treatments in both shoots and roots. Therefore, this study demonstrates the potential of rhizobacteria in alleviating arsenic stress in rice plants.     

固定化混合菌Z1对制革废水Cr3+和OPnEO的同步去除及其响应面法优化

以微紫青霉菌B5和醋酸钙不动杆H1按照1∶1配置成混合菌Z1,并分别以聚乙二醇、改性秸秆、海藻酸钠对混合菌Z1进行包埋,探究其对Cr~(3+)和OPnEO的同步去除处理情况,结果得到OPnEO的去除率分别为53.42%、46.29%、55.14%;而Cr~(3+)的去除率分别达到58.41%、54.24%、60.95%,确定海藻酸钠为Z1的最佳包埋材料.以Cr~(3+)去除率为响应值,利用Plackett-Burman实验确定Cr~(3+)的初始浓度、pH值、温度为影响混合菌Z1去除Cr~(3+)的3个主要因素;运用最陡爬坡实验设计,结合Box-Behnken实验设计及响应面法分析,确定海藻酸钠固定化混合菌Z1在初始pH 7.64、Cr~(3+)初始浓度856.34 mg·L~(-1)、温度30.39℃、OPnEO初始浓度950 mg·L~(-1)、外加2 m L胰蛋白胨条件下培养7 d,Cr~(3+)的去除率达到77.69%,比单因素实验条件下Cr~(3+)的去除率提高7%.     

一株穗花狐尾藻内生菌的分离鉴定及其溶磷特征研究

通过组织表面消毒、切面培养和共生培养的方法从健康的穗花狐尾藻植株体内分离获得了一株具有较强溶磷能力的内生菌,系统发育分析和VITEK 2系统鉴定结果表明,内生菌与沙雷氏菌的亲缘关系最近,因此,命名为Serratia sp.SP5.纯培养实验表明,菌株SP5通过产生酸性物质溶磷,在培养96 h后溶磷能力达到最大,溶解性磷酸根的浓度高达269.60 mg·L-1,培养液的p H值下降至4.32.氮源通过降低菌株SP5的产酸能力而间接影响了其对磷的释放能力,菌株SP5溶磷的氮源优先利用顺序为铵根离子硝酸根离子尿素.适宜菌株SP5生长的p H值范围为5~9,酸性条件下菌株的溶磷能力大于碱性条件,而在p H值为7时与对照相比菌株对磷的相对释放量最大.p H值达到9时,菌株无明显溶磷能力.共存高浓度的金属离子时菌株SP5的溶磷效果明显降低,影响程度的大小顺序为MnFeMg.菌株SP5溶磷的最佳碳氮比为35∶1,在相同碳氮比下,不同的碳氮量会明显影响溶磷效果.底泥模拟培养条件下,菌株SP5可通过释放底泥中的闭蓄态磷(O-P)和钙磷(Ca-P)而明显提高水相中的磷酸根离子浓度,与对照相比,底泥中这两种磷形态分别下降了15.21%和34.87%,而水相中溶解性磷酸根离子是对照的1.5~2.0倍.研究结果为研究富营养化水体中磷的生物地球化学过程提供了新的科学数据.     

饥饿对两级SBR反应器内活性污泥的影响

为探究饥饿期内活性污泥中微生物活性的衰减速率、物理性质的变化及恢复情况,研究了好氧〔ρ(DO)=5.50 mg/L〕/缺氧〔ρ(DO)≤0.03 mg/L〕饥饿期对稳定运行的两级SBR反应器内污泥活性的影响,同时考察了污泥的恢复能力及理化性质的变化. 结果表明:30 d的饥饿使COD_Cr去除率下降了20.8%,r_COD(异养菌活性)由111.8 mg/(L·h)降至59.2 mg/(L·h);NH₄+-N去除率下降了59.2%;COD_Cr去除率、r_COD、NH₄+-N去除率分别在94、97、95 d时恢复. 饥饿期内AOB(氨氧化细菌)的衰减速率(k_AOB)为0.029 d-1,NOB(亚硝酸盐氧化细菌)的衰减速率(k_NOB)为0.021 d-1. 恢复初期AOB的恢复速率(k_AOB′=0.125 d-1)大于NOB的恢复速率(k_NOB′=0.069 d-1),导致NO₂--N的累积. 此外,饥饿期ρ(MLSS)下降了42.6%;SVI(污泥指数)由71.2 mL/g升至135.1 mL/g;w(EPS)(EPS为胞外聚合物)由37.9 mg/g(以VSS计)降至18.5 mg/g,其中主要由w(PN)(PN为蛋白质)的减少所致;各物理指标的变化均可在恢复期内恢复.      

以SRB颗粒污泥为载体的硫酸盐型厌氧氨氧化的启动研究

采用复合式厌氧折流板反应器(HABR),研究先驯化硫酸盐还原菌(SRB)颗粒污泥、再以之为载体进行硫酸盐型厌氧氨氧化的启动,通过NH_4~+-N、SO_4~(2-)、COD等指标的变化探讨启动的效能。在7pH8.5、温度为(32±1)℃的条件下,采用低负荷启动方式,以CH_3COONa为有机碳源,通过逐步缩短HRT提高进水负荷来驯化培养硫酸盐还原菌颗粒污泥。结果表明,SO_4~(2-)与COD去除效果逐步达到稳定,最高去除率分别为86.2%和68.8%,S0全程积累并趋于稳定,经过60 d的驯化,SRB颗粒污泥平均粒径达到3 mm,硫酸盐还原反应启动成功。之后以驯化成熟的SRB颗粒污泥为载体,保持COD为50 mg/L,通过提高进水中NH+4-N和SO2-4负荷的方式启动硫酸盐型厌氧氨氧化。结果表明,NH_4~+-N和SO_4~(2-)去除效果逐步上升并稳定在50%以上,最高分别达到52.5%与53.7%。硫酸盐型厌氧氨氧化成功启动。     

底泥中红霉素耐受菌群的多样性分析

为了正确评估抗生素残留的环境风险,了解长期受低浓度红霉素污染条件下,水体底泥中红霉素耐受菌群结构的变化特点,采用传统的微生物培养法和分子生物学方法对长期受低浓度红霉素污染的水体底泥中红霉素耐受菌群结构的多样性进行了分析.结果表明:采用传统的培养方法,从底泥中可分离筛选出3株对红霉素有耐受能力的菌株,根据其形态学特征及16S rDNA序列分析,初步鉴定为赖氨酸芽孢杆菌(Lysinibacillussp.)、土壤芽孢杆菌(Solibacillus silvestris)以及蜡样芽孢杆菌(Bacillus cereus),其中敏感菌蜡样芽孢杆菌经低浓度红霉素诱导对红霉素表现出一定的耐药性.进一步构建底泥耐药菌群16S rDNA克隆文库发现,底泥中红霉素耐受菌群可分为三大类群,其中未获培养的细菌(Uncultured bacterium)在整个文库中比例最大,占65.06%,其次为芽孢杆菌纲(Bacilli)和梭菌纲(Clostridia),分别占文库比例33.73%和1.20%.文库中可培养的耐药菌优势类群为芽孢杆菌纲,与传统培养方法得到的结果相一致.上述研究表明,在长期受低浓度红霉素污染的环境中,红霉素耐药菌的存在具有一定的广泛性和潜在性,将可能威胁人类健康和生态系统,应当重视其生态风险评价与管理.     

UV light tolerance and reactivation potential of tetracycline-resistant bacteria from secondary effluents of a wastewater treatment plant

Tetracycline-resistant bacteria (TRB) are of concern as emerging microbial contaminants in reclaimed water. To understand the effects of UV disinfection on TRB, both inactivation and reactivation profiles of TRB, as well as 16 tetracycline-resistant isolates from secondary effluent, were characterized in this study. The inactivation ratio of TRB was significantly lower (3.0-log) than that of heterotrophic bacteria (> 4.0-log) in the secondary effluent. Additionally, the proportion of TRB significantly increased from 1.65% to 15.51% under 20 mJ/cm² ultraviolet (UV) exposure. The inactivation rates of tetracycline-resistant isolates ranged from 0.57/s to 1.04/s, of which tetracycline-resistant Enterobacter-1 was the most tolerant to UV light. The reactivation of TRB, tetracycline-resistant isolated strains, as well as heterotrophic bacteria commonly occurred in the secondary effluent even after 20 mJ/cm² UV exposure. The colony forming ability of TRB and heterotrophic bacteria reached 3.2-log and 3.0-log under 20 mJ/cm² UV exposure after 22 hr incubation. The final inactivation ratio of tetracycline-resistant Enterobacter-1 was 1.18-log under 20 mJ/cm² UV exposure after 22 hr incubation, which is similar to those of TRB (1.18-log) and heterotrophic bacteria (1.19-log). The increased proportion of TRB and the reactivation of tetracycline-resistant enterobacteria in reclaimed water could induce a microbial health risk during wastewater reuse.     

固定化SRB处理低浓度含铬废水

主要选用海藻酸钠和聚乙烯醇等为包埋剂,采用包埋法对SRB细菌进行固定化,并且以固定化小球对铬的去除率为主要参考指标,从失重率、传质性等方面综合考虑,通过正交试验确定固定化小球的最佳配比,同时采用这种新型的固定化小球处理含低浓度铬废水。试验结果表明,固定化SRB小球的最佳包埋条件为:聚乙烯醇8%,二氧化硅2%,海藻酸钠0.2%,活性炭3%,菌液含量30%,饱和硼酸中氯化钙2%,交联时间24 h。固定化小球处理低浓度含铬废水的最佳条件为:p H为6,温度为30℃,初始铬离子浓度为1 mg/L,在250 min内小球表面的活性位点趋于饱和,此时Cr去除率为92%。     

反硝化聚磷菌的筛选及脱氮除磷特性

为进一步研究反硝化聚磷菌的机理和性能,对经过SBR反应器富集驯化后的污泥进行实验研究。采用吸磷实验、硝酸盐还原产气实验、革兰氏染色及异染颗粒染色等方法,筛选得到N10、N17和N20 3株菌株;经过16S r DNA的测序鉴定,建立系统发育树,确定3株菌株均为气单胞菌属。绘制3株菌株的生长曲线,并进行厌氧/缺氧和厌氧/好氧静态实验,发现N10和N17菌株在两组实验中均具有较好的脱氮除磷效果,O2和硝酸盐均可作为电子受体;N20菌株仅在厌氧/缺氧实验中表现出脱氮除磷效果,仅可利用硝酸盐作为电子受体。