一株高效邻苯二甲酸二丁酯降解菌的筛选、鉴定及其降解特性研究

从活性污泥中分离出1株以邻苯二甲酸二丁酯(DBP)为碳源和能源生长的高效降解菌DP-2,经形态观察、生化鉴定及16S rDNA序列分析,鉴定该菌株为不动杆菌(Acinetobacter sp.).采用单因素试验研究了不同试验条件(接种量、DBP浓度、NaCl浓度和碳源)对菌株DBP降解特性的影响,结果表明:接种量大于10%时,菌株DP-2在3 d内对初始浓度为10 mg·L~(-1)的DBP降解率可达到90%以上;DBP初始浓度为5—50 mg·L~(-1)时,菌株在6 d内对DBP降解率均能达到90%以上,但高浓度DBP会影响菌株DP-2生长,DBP浓度为1000 mg·L~(-1)时,DBP降解率仅为26.88%;菌株降解DBP的最佳NaCl浓度范围为0—20 g·L~(-1);此外,醋酸钠、蔗糖、葡萄糖添加对于菌株降解DBP均有一定的促进作用,其中葡萄糖效果最为明显.在此基础上,采用响应曲面法优化了菌株降解DBP的培养条件并进行了试验验证,在盐度为5 g·L~(-1),接种量为17.14%,底物浓度为9.81 mg·L~(-1),菌株对DBP的降解率为85.86%.     

测定无机阴离子和低分子量有机酸的新型阴离子交换柱

AS1 8柱是氢氧化物选择性阴离子交换柱 ,用于测定无机阴离子和低分子量的有机酸 ,包括F- ,乙酸根 ,甲酸根 ,Cl- ,NO-2 ,SO2 -4 和PO3-4 等 .AS1 8柱可用于氢氧化物等度淋洗或梯度淋洗 .建议与氢氧化钾淋洗液在线淋发生器EG40或EG5 0结合使用 .AS1 8柱的高容量和选择性为优化阴离子的分离提供了灵活性 .AS1 8柱用于以氢氧化物为淋洗液 ,等度快速分离简单基体样品中的常见无机阴离子是很理想的 .AS1 8柱容量高 ,可使用氢氧化物梯度淋洗液以及大体积进样来测定复杂基体样品中低浓度级别的无机阴离子 ,包括饮用水和废水 .美国EPA水质…     

饮用水氯胺法消毒过程中-氯胺的水杨酸分光光度法测定

本文建市了一种水杨酸分光光度法,用于一氯胺的测定.结果表明,在水杨酸为1.0ml,硝普钠为0.10ml,pH值为11.8和反应时间为60min时,方法效果最好,吸光度与一氯胺具有很好的线性关系(R2=0.9999).一氯胺的回收率为100±5%.水中其它共存离子,如SO,,4>2-,PO43-,ca2+,Mg2+等在常见浓度范围内均不影响一氯胺的定量.方法检出限为0.02mg·1-1,对不同浓度标样的测定表明,相对标准偏差<10%,自来水本底中的加标回收率为96.0%-103.5%.     

硫氧化细菌的分离鉴定及降解特性

从浙江华海药业污水处理系统中分离得到一株硫氧化细菌T3,基于形态特征、生理生化、16S rRNA基因序列系统学分析和Biolog鉴定系统分析,鉴定该菌株为根瘤菌属.摇瓶实验结果表明,T3生物降解最适生长温度为30℃,最适pH值为8.0,外加氯化铵、碳源对菌株生长及硫化钠降解有促进作用,驯化后的硫氧化细菌对硫化钠有很强的耐受能力,最优生长条件下,2 d内菌株T3能将400 mg/L以下浓度的硫化钠降解彻底,是一株有应用前景的硫氧化细菌.通过测定代谢过程中各种物质的含量,确定该菌株对硫化钠的去除机理为S2-→S2O32-/S0→SO32-→SO42-.图8表1参18     

饮用水氯胺法消毒过程中一氯胺的水杨酸分光光度法测定

本文建立了一种水杨酸分光光度法,用于一氯胺的测定.结果表明,在水杨酸为1.0ml,硝普钠为0.10ml,pH值为11.8和反应时间为60min时,方法效果最好,吸光度与一氯胺具有很好的线性关系(R~2=0.9999).一氯胺的回收率为100±5%.水中其它共存离子,如SO_4~(2-),PO_4~(3-),Ca(2 ),Mg(2 )等在常见浓度范围内均不影响一氯胺的定量.方法检出限为0.02mg·l~(-1),对不同浓度标样的测定表明,相对标准偏差<10%,自来水本底中的加标回收率为96.0%-103.5%.     

天祝高寒草地三线镰孢的分离鉴定及β-tubulin基因序列分析

按30 m的梯度将海拔2 880～3 360 m分为16个采样点,在同一海拔高度按照"Z"字形取样,共取5个点,对16个采样点的土样混匀后以根残体分离法在镰刀菌选择性培养基上进行分离,对分离得到的镰刀菌菌落进行纯化和单孢分离后,以形态学为基础,参照Nelson分类系统进行鉴定.结果表明:在分离到的212个镰刀菌菌株中,有5个菌株为三线镰孢(Fusarium tricinctum).随机选取2株三线镰孢菌株进行β-tubulin基因序列分析,将PCR产物回收测序后在Genbank上比对,菌株TZh-4-3-7和TZh-11-5-2与GenBank上登记的6个新西兰菌株的亲缘关系最近,同源性达96%.利用DNAStar软件绘制其系统发育树状图,结果显示,菌株TZh-4-3-7和TZh-11-5-2与以上6个F.tricinctum菌株均位于系统发育树的同一分支,聚为一类.本研究采用的β-tubulin基因序列分析结果验证了利用形态学鉴定镰刀菌的准确性;将β-tubulin基因序列分析应用于三线镰孢的鉴定中,拓宽了鉴定途径,提高了鉴定的准确性;并了解了三线镰孢在天祝高寒草原不同海拔高度的分布,丰富了我国草原镰刀菌资源库,为进一步探讨镰刀菌与草原退化的关系奠定了基础.     

极端嗜热厌氧纤维素菌的分离鉴定、系统发育分析及其酶学性质的研究

从云南高温温泉、油井等热源地区采集的大量样品中,获得了一株特殊的极端嗜热厌氧纤维素分解菌B2.分离菌株直杆,革兰氏阴性(G-),未观察到孢子,细胞单个或成对出现.菌体大小为0.4μm×(2-4)μm,严格厌氧,生长温度范围50-70℃,最适生长温度65℃.pH范围4-8,最适pH 7.0.在纤维素粉琼脂上菌落直径2-4 mm,乳白色.分离菌株能利用纤维二糖、葡萄糖、蔗糖、松子糖、淀粉、覃糖等作为碳源,分离菌株还可利用纤维素滤纸、纤维素粉、微晶纤维素、纤维素粉MN300和甘蔗渣、水稻秸杆.发酵纤维素产生乙醇、乙酸.在菌株B2的纤维素酶系中,C1酶、Cx酶和β-葡萄糖苷酶的最适温度分别为80℃、80℃和70℃,其比值为1:9:10,同时发现Cx酶具有较高的热稳定性.部分长度的16S rDNA序列分析表明,分离菌株B2与Thermoanaerobacter ethanalicus具有99.8％相似性.分离菌株B2为Thermoanaerobacter属.图5表3参21     

温度和溶菌酶敏感菌株的L-谷氨酸合成

用温度敏感型菌株发酵生产L-谷氨酸不存在生物素亚适量问题,因此该方法在国际上被广泛使用.通常采用对出发菌株进行传统诱变的方法获得温度敏感型菌株.以谷氨酸棒杆菌CICC 10226为出发菌株,先克隆其ItsA基因,然后通过基因敲除的方法构建了突变菌株Corynebacterium glutamicum WT ΔL,该菌株同时具有温度敏感性和溶菌酶敏感性.经透射式电子显微镜观察发现,于38℃培养的突变株细胞与在30℃培养的同一种细胞相比,细胞明显增大,而出发菌株无该现象.发酵试验表明,在生物素过量的情况下,在发酵进入细胞产酸期后通过将发酵温度从原来的30℃提高到38℃,温度敏感突变株的产酸量增加近5倍.如果在发酵培养基巾添加适量的琥珀酸和乙酸,该菌株与不添加的在30℃培养的对照相比,产酸量增加近6.5倍.对于野生型出发菌株而言,在生物素过量的情况下,无论是否采用变温发酵方法都几乎不产酸.说明温度敏感突变株即使在生物素过量的情况下也能通过变温发酵诱导其合成并分泌产生L-谷氨酸.图10表1参13     

苯甲酸类化合物的微生物降解研究

用富集培养法从工业污水中分离到5个能以苯甲酸为唯一碳源和能源而生长的细苗菌株：不动杆菌（Acinetobactersp.）BJ1，无色杆菌（Achromobactersp.）BY1，假单胞苗（Pseudomonasspp.）SJ1、SY1和SH1、测定了这5个菌株的底物特异性和抗菌素抗性其中BJ1菌株含有1个大质粒和2个小质位在最适培养条件下BJ1菌株对苯甲酸的降解率达98％以上．     

一株耐盐嗜热菌Aneurinibacillus thermoaerophilus H7的分离及其油脂降解特性

针对高盐、高油餐厨垃圾高温堆肥功能菌株缺乏的问题,以大豆油为唯一碳源,通过测定生物量、脂肪酶活性和油脂降解率,从餐厨垃圾堆积处的土壤样品中分离筛选出一株嗜热油脂降解菌H7.通过形态特征、生理生化特征和16SrDNA序列分析,对筛选的菌株进行鉴定,考察其耐盐能力、油脂降解和生长特性.结果表明,菌株H7为嗜热嗜气解硫胺素芽孢杆菌(Aneurinibacillus thermoaerophilus),最高耐盐浓度为30 g/L.菌株H7在油脂浓度为15 g/L的发酵培养基中发酵72 h,油脂降解率为60.11%,菌体浓度OD600为1.88,脂肪酶活性为11.65 U/mL.菌株H7可生长的温度为40-60℃,pH值为5-8,摇床转速为120-240 r/min,最适生长的温度为50℃,pH值为6,摇床转速为220 r/min.本研究获得了具有耐盐性和能降解高浓度油脂的嗜热菌株H7,可为高盐高油脂含量的餐厨垃圾堆肥提供微生物菌种资源.(图8表2参34)     

Bioassay of biotin and its distribution in the sea

A simple, rapid and sensitive method of microbiological assay for the determination of biotin in seawater has been developed by using a marine bacterium, Achromobacter sp. (Strain yH-51) as an assay organism. When the organism was incubated at 25°C for 48 h in a synthetic medium with constant shaking, a standard reference curve of bioassay was obtained. The organism is sensitive in the range 0.1 to 8.0 ng/l biotin. The distribution of biotin has been determined in the East China Sea, the North Pacific Ocean, and the bays and inlets along the Pacific coast of Japan. Biotin concentrations in surface waters were in the range from less than 0.1 ng/l up to 57.9 ng/l; generally high in coastal waters and low in deep open waters. The concentrations were generally high in surface waters, and rapidly decreased with depth down to 50 m, below which values were generally low. The vertical distribution of biotin followed closely the concentration of chlorophyll a, with high concentrations of biotin restricted to the euphotic layer of high chlorophyll a contents at most stations observed. Biotin concentrations in surface waters were generally high during summer, but in autumn and winter the concentrations decreased markedly.     

不同植被对工业污染区盐渍土的改良效果

以包头市西郊工业污染区盐渍化弃耕地为对象，研究了不同植被对土壤水盐动态和体积质量的影响。研究表明，造林后使林地内草本植物得到较快恢复，且归还土壤的枯落物比木本植物多，改良盐渍土壤和促进脱盐的作用较好；随植被盖度增加，盐渍土土壤含盐量和体积质量降低，盐渍土性状逐步改善。因此，在实施以耐盐树种造林为先导的工业污染区盐渍土恢复植被工程时，应适当降低造林密度，以利于林地内草本植物的恢复和建立林草复合植被，提高工业污染区盐渍土的生物治理效果。     

芹菜赖氨酸加苏氨酸抗性细胞系的选择

用４ｍｍｏｌ－／Ｌ浓度的赖氨酸加苏氨酸作为选择压，经相同浓度液体和固体选择培养基连续选择，从未经诱变处理的芹菜胚性愈伤组织筛选出一个稳定的赖氨酸加苏氨酸抗性细胞系并得到再生植株。抗性细胞系再生植株整株的氨基酸含量比对照组植株高１０倍左右。但是地上部分氨基酸含量，抗性细胞系再生植株比对照植株有所降低。两种植株叶片的细胞色素氧化酶同工酶谱有明显差异，抗性细胞系再生植株有４条谱带，而对照植株只有两条谱带     

Biologische Tests von Bodenextrakten zur Erkennung von Bodenkontaminationen Einstiegswerte und Test-Fensterbreite

Initial values and windows of competence together with a biological test system are introduced to identify soil contaminations. Theinitial values are defined as the amount of extract equivalent to a defined amount of soil (gramm soil equivalent) and the same amount of test medium. This is equivalent to a complete replacement of the test medium with the soil being tested. Theeffect limits should be at least the double standard deviation of the blank values. Based on the values found when testing uncontaminated soils, a higher threshold value has to be established for some test systems. Should no relationship be found to a standard test medium (e.g. agar as a matrix in the Ames assay), the so-calledwindow of competence is defined. Within this window no natural response is found when ex amining uncontaminated soil. For mutagenicity tests, the double spontaneous reversion rate (reversion coefficient of 2) as related o the blank sample is evaluated as the effect limit.     

人工景观生态湖滨净化带植物的遴选

对水生植物处理生活污水的效果等展开了一些初步的研究。利用盆栽试验,比较了香蒲、美人蕉等8种水生植物的净化效果,所选的8种植物对污水均有不同程度的去除效果。挺水植物对污水的净化效果和耐污能力均优于沉水植物。利用水生植物处理生活污水,5 d的停留时间可以使污水达到较好的去除效果。挺水植物中美人蕉和水竹对污水的净化效果比香蒲和灯心草更好;沉水植物中伊乐藻的净化效果最好。植物对污水的净化效率有明显的季节差异,冬季污水净化效率低于夏秋两季,沉水植物在不同季节脱氮除磷效果有显著的差异。     

6种观赏植物吸收甲醛能力比较研究

城市住宅的室内空气污染十分严重,甲醛已经成为中国目前室内空气中的首要污染物.在已有的报道中,盆栽观赏植物吸收甲醛的效果并未排除盆土的净化能力,由此植物净化效果显然缺乏科学性.选用6种常用室内观赏植物,将盆土与茎叶部分隔开,放入自行研制的甲醛熏蒸箱中,对植物进行熏蒸实验.测得熏蒸前后甲醛的变化量和植物叶面积,以净吸收率及单位叶面积甲醛减少量来比较6种植物吸收甲醛能力的大小.主要结论如下:(1)甲醛质量浓度是国际标准(0.08 mg·m~(-3))23倍情况下,6种观赏植物均能吸收空气中的甲醛,且因种类不同吸收能力大小不一.吸收甲醛能力排序为:广东万年青(Aglaonema modestum)>绿萝(Epipremnum aureum)>垂叶榕(Ficus benjamina)>虎尾兰(Sansevieria trifasciata)>龟背竹(Monstera deliciosa)>四季秋海棠(Begonia semperflorens-hybr.).单位叶面积植物吸收甲醛量依次为:广东万年青绿萝>虎尾兰>龟背竹>垂叶榕>四季秋海棠.(2)甲醛质量浓度是国际标准(0.08mg·m~(-3))57倍情况下,6种观赏植物吸收甲醛能力排序为:垂叶榕>虎尾兰>绿萝>广东万年青>龟背竹>四季秋海棠.单位叶面积植物吸收甲醛量依次为:虎尾兰>垂叶榕>龟背竹>广东万年青>绿萝>四季秋海棠.(3)根据净吸收率及单位叶面积吸收甲醛量的差异,将植物的吸收能力分为两类,吸收甲醛能力较强的植物有:广东万年青、绿萝、虎尾兰、龟背竹、垂叶榕(与空白组相比有显著性差异(P<0.05));较低的植物有:四季秋海棠(与空白组相比无显著性差异).     

Applying chemical sedimentation process in drinking water treatment plant to address the emergent arsenic spills in water sources

Arsenic (As) spills occurred more frequently and sometimes polluted water sources in recent years in China. It is as urgent need to develop emergency treatment technologies to address the arsenic threat for large-scale water treatment plants. In response, we developed a chemical sedimentation technology to remove arsenic contaminants for water treatment plants. Bench-scale experiments were conducted to investigate the efficiency of arsenic removal and the influencing factors of the chemical sedimentation treatment process. The influencing factors included the choice and dosage of coagulants, the valence of arsenic and pH value of solution. The As(V) contaminants can be almost completely removed by ferric or alum coagulants. The As(III) contaminants are more recalcitrant to chemical sedimentation, 75% for ferric coagulant and 40% for alum coagulant. The quantitative results of arsenic removal load by different ferric or alum coagulants were presented to help determine the parameters for arsenic treatment technology. The dominant mechanism for arsenic removal is static combination, or adsorption of negative arsenic species onto positive ferric hydroxide or alum hydroxide flocs. The efficiency of this treatment technology has also been demonstrated by a real production test in one water treatment plant with arsenic-rich source water and one emergency response. This technology was verified to be quick to set-up, easy to operate and highly efficient even for high concentration of arsenic.     

用改进的TRAP法测定树木端粒酶活性

TRAP法是一种常用的测定端粒酶活性的方法,但并不太适合测定木本植物的端粒酶活性.因此以油松针叶为实验材料,对TRAP法进行改进.参照前人的研究通过对比实验证明,在端粒酶提取过程中将PEG8000加入上清液,可获得高效的端粒酶.在此基础上,对PCR先导引选择TS、模板浓度选择100 ng,最终获得油松针叶高质量的端粒酶PCR产物.应用SYBR Green I替代银染液对电泳凝胶进行染色,获得条带清晰、重复性好的电泳结果.通过对油松针叶、银杏叶片、胡杨愈伤组织、沙冬青悬浮细胞等4种木本植物实验材料进行测定,证明该改进的技术体系可能是适合于木本植物端粒酶活性测定的稳定的、灵敏度高的可行方法.图5表1参17     

Antimicrobial activities of extracts from tropical Atlantic marine plants against marine pathogens and saprophytes

Studies investigating disease resistance in marine plants have indicated that secondary metabolites may have important defensive functions against harmful marine microorganisms. The goal of this study was to systematically screen extracts from marine plants for antimicrobial effects against marine pathogens and saprophytes. Lipophilic and hydrophilic extracts from species of 49 marine algae and 3 seagrasses collected in the tropical Atlantic were screened for antimicrobial activity against five ecologically relevant marine microorganisms from three separate kingdoms. These assay microbes consisted of the pathogenic fungus Lindra thalassiae, the saprophytic fungus Dendryphiella salina, the saprophytic stramenopiles, Halophytophthora spinosa and Schizochytrium aggregatum, and the pathogenic bacterium Pseudoaltermonas bacteriolytica. Overall, 90% of all species surveyed yielded extracts that were active against one or more, and 77% yielded extracts that were active against two or more assay microorganisms. Broad-spectrum activity against three or four assay microorganisms was observed in the extracts from 48 and 27% of all species, respectively. The green algae Halimeda copiosa and Penicillus capitatus (Chlorophyta) were the only species to yield extracts active against all assay microorganisms. Among all assay microorganisms, both fungi were the most resistant to the extracts tested, with less than 21% of all extracts inhibiting the growth of either L. thalassiae or D. salina. In contrast, over half of all lipophylic extracts were active against the stramenopiles H. spinosa and S. aggregatum, and the bacterium P. bacteriolytica. Growth sensitivity to hydrophilic extracts varied considerably between individual assay microorganisms. While 48% of all hydrophilic extracts were active against H. spinosa, 27% were active against P. bacteriolytica, and only 14% were active against S. aggregatum. Overall, more lipophilic extracts inhibited microbial growth than hydrophilic extracts. The variability observed in the antimicrobial effects of individual extracts against each assay microorganism reflects the importance of choosing appropriate test microbes in assays from which ecologically relevant information is sought. Results from this survey demonstrate that antimicrobial activities are prevalent among extracts from marine algae and seagrasses, suggesting that antimicrobial chemical defenses are widespread among marine plants.     

人工湿地植物量及其对净化效果影响的研究

人工湿地污水处理系统中,植物对污水净化效果起着非常重要的作用,而植物量是植物生长的关键参数之一。为了明确植物量对污水处理效果的影响,在2002—2008年期间,详细测定了芦苇(Ph.australis Trin)、再力花(Thalia dealbata)、荻(M.sacchariflorus)和美人蕉(Canna indica)4种典型湿地植物不同年度的生物量、不同生长阶段的生物量、生物量年产率,以及同期湿地COD、BOD5、TN和TP等的去除率。结果表明:湿地植物在栽植成活、稳定生长以后,至少要再经过1个以上的生长周期才能完全适应湿地环境,达到生物量的最大值,其中芦苇根系和茎叶生物量显著高于其他植物;同种植物的植物量与COD、BOD5、TN去除率呈显著正相关,与TP相关性不强;植物量随生长周期的变化对湿地净化能力的影响显著,确定合理收割期有利于湿地的稳定运行;不同种类植物生物量年产率与各种污染物的去除率均显著相关,芦苇、再力花和美人蕉均具有较高的生物量年产率,污染物去除能力较好。在试验范围内,芦苇湿地的运行稳定性最好,再力花湿地具有最强的脱氮能力,美人蕉湿地能快速形成规模、实现稳定运行,这些为构建不同特点和不同需要的湿地提供了重要参考。