五株微囊藻毒素降解菌的分离与鉴定

有毒蓝藻产生的蓝藻毒素导致地表水的污染已是全球普遍的现象,最常见的毒素是能导致肝癌的肝毒素微囊藻毒素(MC),迄今为止尚无适用的MC驱除技术,微生物降解MC可能是一条有效的途径．本研究以铜绿微囊藻(Microcystis aeruginosa PCC 7806)所产生的MC为底物,从多个形成微囊藻水华的水体中分离获得能显著降解MC的细菌18株,相对降解能力从18．4％到48．2％．对其中5株MC相对降解能力较强的菌,通过形态特征和生理生化特征分析,鉴定出5株菌分别为:不动细菌属(Acinetobacter)、肠杆菌属(Enterobacter)、微杆菌属(Microbacterium)、芽孢杆菌属(Bacillus)和弗拉特氏菌属(Frateuria),均不同于已报道的属种．     

太湖微囊藻毒素时空分布特征及与环境因子的关系

采用高效液相色谱法(HPLC)对太湖水体中溶解态微囊藻毒素(MC-LR;MC-RR)浓度进行检测,探讨微囊藻毒素浓度的时空变化及其与氮、磷、总有机碳、蓝藻生物量以及温度等湖泊环境因子之间的关系.结果表明:微囊藻毒素表底层浓度基本一致;在一天之内的变化没有明显规律;冬季微囊藻毒素浓度较高;位于竺山湾和西部沿岸地区的两个点...     

2013年洱海水华期间微囊藻毒素和浮游植物动态变化

2013年秋季,洱海再次出现大规模水华,周边几十万居民的饮用水安全引起当地政府的极大关注.为了解水华发生期间洱海水体微囊藻毒素的含量和浮游植物的种类及丰度,分别在2013年9月和10月两次在洱海采集水样,测定其中两种主要微囊藻毒素MC-LR和MC-RR含量,探讨微囊藻毒素含量对洱海周边水源地的安全影响.同时也对水华期间的浮游植物特别是蓝藻进行研究,从而分析微囊藻毒素与浮游植物种类和丰度的关系.结果表明,洱海水体中微囊藻毒素含量偏低,暂不会对人类健康产生威胁,但在洱海的水鸟体内仍然检测到微囊藻毒素的富集,显示洱海蓝藻水华对水生生物的潜在危害不容忽视.浮游植物的研究发现,洱海水华的优势种属为微囊藻(Microcystis)和暗丝藻(Psephonema aenigmaticus).9月份暗丝藻更占优势,10月份则是微囊藻更占优势.研究结果显示处于富营养化初期的洱海水体中湖湾微囊藻毒素的含量与浮游植物中产毒蓝藻的种类和丰度密切相关.     

巢湖微囊藻毒素异构体组成的时空分布特征及影响因子

有害蓝藻释放微囊藻毒素（MCs）,严重威胁饮用水源地用水安全.为了解巢湖MCs污染状况及其异构体组成对水质的影响,于2012年夏季（8月）和秋季（11月）,2013年冬季（2月）和春季（5月）进行采样分析,研究了巢湖水体中胞内微囊藻毒素（IMCs）和胞外微囊藻毒素（EMCs）异构体的时空分布及其与环境因子的关系.结果发现,IMCs和EMCs的平均浓度变化范围分别为0.12~6.45 μg/L和0.69~1.92 μg/L.在3种常见的异构体中,MC-RR和MC-LR比例较高,MC-YR最低,MC-RR和MC-LR是巢湖水体中MCs的主要异构体类型.IMCs和EMCs的异构体浓度及其比例呈现不同的时空分布特征.微囊藻生物量、水温、总磷浓度是影响IMCs和EMCs异构体浓度及其组成变化的关键环境因子.本研究表明巢湖富营养化严重的西湖区夏季能合成更多的MC-RR异构体,而秋、冬季节偏向于释放生理毒性更强的MC-LR异构体.了解MCs异构体组成变化及其关键影响因素,有助于预测预警水体MCs污染状况和评估饮用水源地MCs风险.     

铜绿微囊藻(Microcystis aeruginosa)次生代谢物对普通小球藻(Chlorella vulgaris)生长及有效量子产率的影响

浮游植物间的交互化感作用被认为是自然水体中浮游生物群落演替及优势种转换的主要因素之一.铜绿微囊藻(Microcystis aeruginosa)是富营养化湖泊中的常见蓝藻,其产毒品系相较非产毒品系具有较高的竞争优势,其产生的主要次生代谢产物微囊藻毒素-LR(MC-LR)通常被认为是重要的化感物质.但是到目前为止其在水生生态系中的生物学功能尚不明确.为探究并区分MC-LR及其他次生代谢产物对浮游植物的化感效应,本文研究了MC-LR与高浓度普通小球藻(Chlorella vulgaris)生长及有效量子产率的剂量—效应关系;比较了能够产生显著抑制效应的MC-LR、能够产生同等浓度MC-LR的产毒品系铜绿微囊藻滤液以及细胞粗提液对小球藻生长及有效量子产率的抑制效应.结果显示,MC-LR能够对小球藻生长及有效量子产率产生抑制作用,并且抑制率随着MC-LR浓度的增加而增大.在用200μg/L及以上浓度的MC-LR感染24 h后,叶绿素a浓度为1500μg/L的小球藻的生长与有效量子产率均比对照组显著降低.能够产生相同浓度MC-LR的产毒品系铜绿微囊藻细胞滤液对其没有产生显著影响,而其细胞粗提液却能够产生更强的抑制效应.与MCLR处理组相比,粗提液处理组小球藻的生长及有效量子产率分别降低了14%和3%.以上结果表明,MC-LR对普通小球藻的化感作用具有剂量依赖性;MC-LR之外的其他次生代谢物也能够产生抑制效应.     

微囊藻毒素生物学功能研究进展

我国淡水水体蓝藻水华的频繁发生已成为我国目前和今后长时期内面临的重大水环境问题.微囊藻(Microcystis)因其产生对人类健康具有危害的微囊藻毒素而尤其受到重视.研究者对构成蓝藻水华的主要种类、微囊藻毒素的毒性、毒理等方面的认识逐渐明晰,但对微囊藻毒素生物学功能的了解还相对较少.本文对微囊藻毒素的产生,尤其是近年来随着技术的发展在微囊藻毒素生物学功能方面的研究开展讨论:总结了微囊藻毒素可能作为化感物质、参与光合作用、在微囊藻的越冬或复苏中可能发挥的作用以及可能参与微囊藻群体形成及维持等4个方面的生物学功能;分析了微囊藻毒素作为信号分子的研究现状;探讨了全球气候变化及实验室模拟条件下微囊藻毒素产毒与无毒株的竞争,并对微囊藻毒素未来的研究热点及组学技术可能在其研究中的应用进行了展望.     

氮及氮磷比对附着藻类及浮游藻类的影响

2005年5月至11月,在河北省YH水库选取5个具有代表性的样点,进行了微囊藻毒素-LR的调查,同时,测定了相应各样点的TN,TP,NH4-N,NO3-N和PO4-P.结果显示,水库中微囊藻毒素-LR(MC-LR)随季节发生变化,其中,7-10月期间相对较高,且大部分超过了生活饮用水地表水标准限值(1 μg/L);MC-LR与水体中的N、P之间的相互关系表明,5-7月期间,水库MC-LR与TP呈正相关,与NH4-N和TN/TP呈负相关;8-9月期间,水库中TP逐渐降低,但其它降低速率低于TN,造成TN/TP明显降低,MC-LR与TP和TN/TP呈正相关,与NH4-N呈负相关;10-11月期间,大量藻细胞死亡,释放到水体中MC-LR也逐渐下降,这时,大量外源营养盐也进入水库,造成微囊藻毒素-LR的变化与TN、NH4-N、NO3-N呈显著或极显著负相关.这说明在不同季节下,微囊藻毒素-LR与营养盐的关系不同,必须视实际情况而定.     

水华蓝藻产毒的生物学机制及毒素的环境归趋研究进展

本文介绍并评述了蓝藻水华中最常见的毒素——微囊藻毒素的产生途径和环境归趋的国内外研究进展.主要内容包括:微囊藻毒素的来源、结构和一般特性;微囊藻毒素的分子合成机制、分布、产生规律及其功能;以及微囊藻毒素的环境归趋.重点介绍了在毒素环境归趋研究方面的重要突破,指出了该领域研究中存在的问题和今后研究的重点方向.     

微囊藻毒素和温度对萼花臂尾轮虫(Brachionus calyciflorus)形态学特征的影响

近年来,随着全球变暖和水体富营养化程度的日趋严重,淡水湖泊藻类水华现象频发;由此引发的微囊藻毒素对水生生物的生态毒性效应备受关注.本实验探究了4个温度(16、20、24和28℃)和3个微囊藻毒素MC-LR浓度(2.0、4.0和6.0μg/ml)对4个品系萼花臂尾轮虫(Brachionus calyciflorus)形态特征的影响.通过2种DNA分类方法——自动条形码查询系统和Yule溯祖模型将萼花臂尾轮虫划分为3个进化种;4个品系轮虫的形态学参数总体随温度升高呈减小趋势.在16℃下,轮虫的形态特征在各品系间未见显著差异;20℃下的昆明品系KMC23形态参数最小,而西宁XN2品系最大;在24℃下,西宁XN2品系的形态参数显著增大;兰州LZB1和西宁XN2品系对温度变化的响应特征相似且在28℃下最小,这可能与两者属于同一进化种有关.4个品系轮虫的卵特征参数对温度变化的响应各有不同.可见,品系和隐种的不同是造成4个轮虫克隆形态学参数对温度差异响应的重要内源性因素.各品系轮虫中,兰州LZB1品系受温度、MC-LR及两者交互作用的影响最为显著,形态可塑性最强.在高温胁迫下,兰州LZB1品系的所有形态学参数均显著减小,但MC-LR浓度对该品系轮虫的形态学参数并未产生显著影响;经低温胁迫后,低浓度的MC-LR没有显著改变兰州LZB1品系轮虫的形态学参数,而高浓度的MC-LR显著降低了轮虫的大部分形态学参数.微囊藻毒素对轮虫形态可塑性的影响受到了内源性因素和外源性因素的综合制约.     

河蟹养殖池塘微囊藻水华毒性及其光合作用活性特征

苏州市吴中区一河蟹养殖池塘在2013年7月和8月发生了严重的微囊藻水华.采用单一和双重PCR扩增微囊藻毒素合成酶基因,用以检测微囊藻水华是否产毒,结果显示为阳性.同时,采用高效液相色谱测定微囊藻水华的毒性大小.结果表明:7月和8月微囊藻水华的胞内微囊藻毒素浓度分别为1.49和0.88μg/L,胞外微囊藻毒素的浓度分别为0.75和1.09μg/L.另外,采用浮游植物荧光仪Phyto-PAM测定河蟹养殖水体形成水华的微囊藻的光合作用活性.结果显示:7月和8月水华微囊藻的最大光量子产量Fv/Fm分别为0.48和0.44,实际光量子产量ΦPSII分别为0.38和0.32,表明形成水华的微囊藻有较高的生长潜力.非光化学荧光淬灭值NPQ分别为0.28和0.36.从快速光响应曲线RLC的特征参数来看,7月水华微囊藻的光合作用活性和光能利用效率高于8月.本研究结果表明,河蟹养殖池塘水体受到微囊藻水华和微囊藻毒素的污染,进而可能对河蟹食品安全构成潜在威胁.     

Bacterial transport experiments in fractured crystalline bedrock

The efficiency of contaminant biodegradation in ground water depends, in part, on the transport properties of the degrading bacteria. Few data exist concerning the transport of bacteria in saturated bedrock, particularly at the field scale. Bacteria and microsphere tracer experiments were conducted in a fractured crystalline bedrock under forced-gradient conditions over a distance of 36 m. Bacteria isolated from the local ground water were chosen on the basis of physicochemical and physiological differences (shape, cell-wall type, motility), and were differentially stained so that their transport behavior could be compared. No two bacterial strains transported in an identical manner, and microspheres produced distinctly different breakthrough curves than bacteria. Although there was insufficient control in this field experiment to completely separate the effects of bacteria shape, reaction to Gram staining, cell size, and motility on transport efficiency, it was observed that (1) the nonmotile, mutant strain exhibited better fractional recovery than the motile parent strain; (2) Gram-negative rod-shaped bacteria exhibited higher fractional recovery relative to the Gram-positive rod-shaped strain of similar size; and (3) coccoidal (spherical-shaped) bacteria transported better than all but one strain of the rod-shaped bacteria. The field experiment must be interpreted in the context of the specific bacterial strains and ground water environment in which they were conducted, but experimental results suggest that minor differences in the physical properties of bacteria can lead to major differences in transport behavior at the field scale.     

太湖梅梁湾水体及沉积物中微囊藻毒素含量垂向分布特征

为探究太湖梅梁湾水体及沉积物中微囊藻毒素（MC-LR、MC-RR、MC-YR）含量的垂向分布特征,于2018年5月采集梅梁湾6个点位表层水、上覆水、混合水、间隙水以及柱状沉积物样品,并采用超高效液相色谱-串联质谱法分析样品中微囊藻毒素的含量.分析结果表明：水体中（表层水、上覆水、混合水以及间隙水）MC-LR、MC-RR、MC-YR的浓度范围分别为11.80~1297.14、2.50~818.40、1.80~176.00 ng/L,表层水、上覆水以及混合水中MC-LR的浓度高于MC-RR和MC-YR,MC-RR和MC-YR之间差别较小,而间隙水中MCs三种异构体浓度大小顺序为：MC-LR > MC-RR > MC-YR;垂向分布上,间隙水中MCs异构体（MC-LR、MC-RR、MC-YR）浓度均远高于表层水、上覆水以及混合水,表层水MCs异构体浓度略高于上覆水,混合水MCs异构体浓度介于表层水和上覆水之间.对沉积物的研究发现,1~10 cm表层沉积物中MC-LR、MC-RR、MC-YR含量范围分别为0.60~26.95、0~0.90、0~8.10 ng/g,且1~10 cm层中MCs三种异构体平均含量大小顺序为：MC-LR > MC-YR > MC-RR,其中MC-LR、MC-RR、MC-YR的检出率分别为100%、70%、92%;垂向分布上,MC-RR含量较低且变化不大,而MC-YR和MC-LR含量均随沉积物深度的增加先升高后降低.相关性分析结果表明,表层水和混合水中MCs与总磷浓度呈显著正相关,而与总氮浓度无显著相关性;上覆水、间隙水以及沉积物中MCs与总氮、总磷浓度均呈显著正相关.     

高原富营养化湖泊杞麓湖微囊藻毒素分布特征、相关因子及其健康风险分析

杞麓湖作为典型的富营养化高原湖泊,其藻华暴发引发的微囊藻毒素(microcystins, MCs)污染问题会危害水生态安全并造成人体健康风险。为了评估杞麓湖流域MCs的秋冬季节污染现状以及健康风险情况,对湖内和7条主要入湖河流入湖口的表层水进行采样检测,分析MCs主要异构体的时空分布特征,研究MCs与水质和浮游植物等环境因子的关系,并通过人体非致癌健康风险评价MCs的风险等级。结果发现,MCs浓度自秋季到冬季呈下降趋势,秋季湖内胞内MCs(IMCs)占比超过99%,冬季IMCs与胞外MCs(EMCs)浓度接近。MC-RR和MC-LR是主要的MCs异构体类型,其中MC-RR在秋季浓度占比高于MC-LR和MC-YR,而冬季MC-LR最高。MCs与微囊藻密度等生物因子呈极显著正相关关系,与TN、TP和NH₃-N营养盐呈显著负相关关系。杞麓湖EMCs污染的风险指数范围为0.004~0.110,处于极低或低风险,冬季风险略高于秋季,因南岸入湖EMCs污染以及湖内IMCs释放带来的风险需要进一步关注。     

蓝藻水华暴发期间太湖贡湖湾某水厂水源水及出厂水中微囊藻毒素污染分析及健康风险评价

水体富营养化导致的有害蓝藻水华仍是目前全世界普遍面临的水环境问题,而有害蓝藻水华所引起的饮用水安全问题亦受到人们的广泛关注.为了解太湖水源地水源水及自来水厂出厂饮用水中微囊藻毒素(MCs)的污染现状,于2014年8月期间对贡湖湾某水厂水源水及出厂水中浮游植物胞内及胞外MCs浓度进行了调查,并同时检测了相关的理化指标.结果表明,水源水中胞内MCs总浓度平均值为7165.5 ng/L,以MC-LR和MC-RR为主,平均浓度分别为3408.7和3398.8 ng/L,其中MC-RR占总MCs比例的平均值为56.1%;而胞外溶解性MCs浓度相对较低,平均浓度为142.6 ng/L,最高浓度仅为512.8 ng/L.水厂出厂水中胞内MCs的检出浓度(平均值为0.77 ng/L)和检出频率都很低,去除率达99.8%以上;而胞外溶解性MCs的检出浓度(平均值为21.71 ng/L)和检出频率相对较高,但浓度仍远低于国家标准1.0μg/L,其去除率相对较低,仅为62.9%~81.8%.数据分析发现,水源水中胞内与胞外MCs浓度之间呈显著正相关,胞内MCs浓度与总氮(TN)浓度、铵态氮(NH+4-N)浓度、总磷(TP)浓度、高锰酸盐指数(CODMn)和浊度呈显著相关,而胞外MCs浓度与TN浓度、TP浓度、CODMn、浊度和叶绿素a浓度呈显著正相关;逐步回归结果显示,TP对胞内MCs浓度变化的解释率最高,而胞外MCs浓度变化主要与胞内MCs浓度相关.最终,通过对出厂饮用水中MCs浓度非致癌风险指数的计算发现,出厂饮用水对人类健康的威胁较小,但致癌风险相对较高.     

Biodegradation by Co‐inoculated Bacteria and Fungi Alleviates Adverse Effects of Red‐S3B on Growth and Nitrogen Uptake of Wheat

Textile wastewater contains huge quantities of nitrogen (N)‐containing azo‐dyes. Irrigation of crops with such wastewater adds toxic dyes into our healthy soils. One of the ways to prevent their entry to soils could be these waters after the dyes' biodegradation. Therefore, the present study was conducted to evaluate the impact of textile dyes on wheat growth, dye degradation efficiency of bacteria‐fungi consortium, and alleviation of dye toxicity in wheat by treatment with microbial consortium. Among dyes, Red‐S3B (3.19% N) was found to be the most toxic to germination and growth of seven‐day‐old wheat seedlings. Shewanella sp. NIAB‐BM15 and Aspergillus terreus NIAB‐FM10 were found to be efficient degraders of Red‐S3B. Their consortium completely decolorized 500 mg L^?1 Red‐S3B within 4 h. Irrigation with Red‐S3B‐contaminated water after treatment with developed consortium increased root length, shoot length, root biomass, and shoot biomass of 30‐day‐old wheat seedlings by 47, 18, 6, and 25%, respectively, than untreated water. Moreover, irrigation after microbial treatment of dye‐contaminated water resulted in 20 and 51% increase in shoot N content and N uptake, respectively, than untreated water. Thus, co‐inoculation of bacteria and fungi could be a useful bioremediation strategy for the treatment of azo‐dye‐polluted water.     

Accumulation and biodegradation of phenanthrene and fluoranthene by the algae enriched from a mangrove aquatic ecosystem

This study focused on the accumulation and biodegradation of two typical polycyclic aromatic hydrocarbons (PAHs), phenanthrene (PHE) and fluoranthene (FLA), by the diatoms enriched from a mangrove aquatic ecosystem in the Jiulong River estuary, China. After separation, purification and culture, Skeletonema costatum (Greville) Cleve and Nitzschia sp. were exposed to different concentrations of PHE, FLA, and a mixture of the two. The results showed that the tolerance of S.costatum to PHE and FLA was greater than that of Nitzschia sp., and that the toxic effect of FLA on S. costatum and Nitzschia sp. was higher than that of PHE. The microalgal species S. costatum and Nitzschia sp. were capable of accumulating and degrading the two typical PAHs simultaneously. The accumulation and degradation abilities of Nitzschia sp. were higher than those of S. costatum. Degradation of FLA by the two algal species was slower, indicating that FLA was a more recalcitrant PAH compound. The microalgal species also showed comparable or higher efficiency in the removal of the PHE-FLA mixture than PHE or FLA singly, suggesting that the presence of one PAH stimulated the degradation of the other.     

Phenanthrene-degrading pathway of Agrobacterium sp. Phx1

The metabolic pathway of phenanthrene-degrading strain Agrobacterium sp. Phx1 was investigated. Phx1 almost was able to transform 100 υg/mL of phenanthrene completely in 1 day in broth media of beef extract-peptone (BP), Luria-Bertani (LB) and mineral salts media (MS), and LB and BP could promote the growth and degradation efficiency of Phx1. The GC-MS was employed to analyze the metabolites of the 1st, 3rd, 7th days of phenanthrene degradation in MS. As a result, the 1-Hydroxy-2-naphthoic acid (1H2N) and 1-naphthol (NOL) were detected in the metabolites of the 1st day. Only NOL was observed on the 3rd day and it disappeared on the 7th day. The accumulated NOL did not pertain to the defined pathway of phenanthrene degradation by bacteria. The further HPLC study confirmed the finding in GC-MS analysis and found the production of catechol (CAT) from o-phthalic acid (OPA) in the phenanthrene metabolizing, which has never been reported in the defined degrading pathways. This production was also evidenced by the production of CAT using OPA as substrate. All of our results showed that the Agrobacterium sp. Phx1 had a novel phenanthrene-degrading pathway.     

固定化氮循环细菌修复城市湖泊水体脱氮效果及N2O排放

利用从镇江金山湖天然水体中筛选分离出的土著氨化、亚硝化、硝化和反硝化细菌,将氮循环细菌固定化后在金山湖示范工程区进行水体脱氮净化效果研究.结果表明,运行一段时间后水质明显得到改善,氨氮、亚硝酸盐氮、硝酸盐氮以及总氮浓度均显著下降,氨氮浓度指标达到国家水质Ⅰ类标准,总氮浓度指标达到水质Ⅱ类标准.同时,对湖泊水体的N_2O气体排放通量进行了检测,结果显示,随着实验的进行,金山湖区的N_2O气体排放通量逐渐升高,4月份的平均值为23,51μg/(m~2·h)、5月份的平均值为29.52μg/(m~2·h)、6月份的平均值为59.10μg/(m~2·h).水质氮素指标以及N_2O气体排放通量数据说明利用微生物固定化载体强化净化技术对城市湖泊水质净化具有显著效果和重要意义.     

Isolation and characterization of alkane degrading bacteria from petroleum reservoir waste water in Iran (Kerman and Tehran provenances)

Petroleum products spill and leakage have become two major environmental challenges in Iran. Sampling was performed in the petroleum reservoir waste water of Tehran and Kerman Provinces of Iran. Alkane degrading bacteria were isolated by enrichment in a Bushnel–Hass medium, with hexadecane as sole source of carbon and energy. The isolated strains were identified by amplification of 16S rDNA gene and sequencing. Specific primers were used for identification of alkane hydroxylase gene. Fifteen alkane degrading bacteria were isolated and 8 strains were selected as powerful degradative bacteria. These 8 strains relate to Rhodococcus jostii, Stenotrophomonas maltophilia, Achromobacter piechaudii, Tsukamurella tyrosinosolvens, Pseudomonas fluorescens, Rhodococcus erythropolis, Stenotrophomonas maltophilia, Pseudomonas aeruginosa genera. The optimum concentration of hexadecane that allowed high growth was 2.5%. Gas chromatography results show that all strains can degrade approximately half of hexadecane in one week of incubation. All of the strains have alkane hydroxylase gene which are important for biodegradation. As a result, this study indicates that there is a high diversity of degradative bacteria in petroleum reservoir waste water in Iran.