微囊藻毒素-LR降解菌的筛选及降解特性研究

从上海市淀山湖表层水体中筛选分离出了1株降解微囊藻毒素-LR(MC-LR)的细菌并研究了其降解特性。根据细胞形态结构、生理生化特征及其16S rDNA基因序列分析,鉴定分离菌株DHU-28(GenBank序列登录号为HM047512)属嗜麦芽寡养单胞菌(Stenotrophomonas maltophilia)。微囊藻毒素降解实验结果表明,该菌株能在以MC-LR为唯一碳源、氮源的无机盐培养基中生长,6 d内可将初始质量浓度为15 mg/L的MC-LR降解为8.12 mg/L,降解效率达到45.9%。菌株DHU-28的最适生长温度是30℃,最适生长pH为7.0。酵母粉、蛋白胨、葡萄糖等营养物质可以明显促进菌株对MC-LR的降解效率,尤其是加入50 mg/L酵母粉后,6 d降解率达到63.2%。     

DSF型群体感应系统的缺失导致嗜根寡养单胞菌溶藻作用的增强

水环境中的微藻与藻际微生物之间关系复杂多变,而微生物的群体感应(QS)作用会影响并调控菌藻的共生关系。以嗜根寡养单胞菌(Stenotrophomonas rhizophila)野生株(WT)和QS扩散信号因子(DSF)合成酶基因-rpfF基因敲除株(△rpfF)为实验菌株,阐明DSF型群体感应是否调控嗜根寡养单胞菌对铜绿微囊藻的溶藻效果。结果表明：嗜根寡养单胞菌野生株和rpfF基因敲除株的溶藻特性相似,均具有针对铜绿微囊藻的溶藻特异性,主要通过分泌溶藻物质间接作用于藻细胞;野生株和敲除株培养48 h后的无菌滤液均有显著的溶藻作用,在投加体积分数10%无菌滤液的条件下,野生株和敲除株7 d溶藻率分别为53%和78%。此外,实验菌株分泌的溶藻活性物质具有热稳定性和酸碱耐受性,且不容易被乙醇沉淀;三维荧光光谱、紫外可见光光谱、傅里叶变换红外光光谱分析表明菌株胞外滤液中主要以类腐殖酸物质为主,芳香化程度较高;通过比较荧光强度和吸光度可知DSF合成酶基因缺失的嗜根寡养单胞菌的无菌滤液中类腐殖酸物质含量显著高于野生株。结果表明,DSF-QS缺失的嗜根寡养单胞菌生物量增多,能分泌更多的溶藻物质,增强...     

一株具有藻毒素降解和溶藻功能细菌的分离鉴定

从太湖水华腐烂蓝藻中富集筛选出一株编号A-8的细菌。该菌株菌落圆形、灰白色,革兰氏染色阴性,氧化酶阴性,接触酶阳性;分子系统进化树显示为溶血不动杆菌。根据以上结果,将此菌株鉴定为溶血不动杆菌。此外,对这株菌降解微囊藻毒素、不同生长时期的细菌的溶藻效果及溶藻专一性进行了研究。结果显示:该菌在3 d时间里将藻毒素降解了30.4%;细菌通过分泌胞外物质溶藻,细菌在衰退期时胞外分泌物的溶藻效果最好;该菌还对瘤状念珠藻、颤藻等9种蓝藻有溶藻作用。     

嗜麦芽寡养单胞菌DHHJ固态发酵羽毛工艺的研究

为了将微生物处理羽毛工艺产业化,利用课题组筛选得到的嗜麦芽寡养单胞菌,在液态发酵羽毛工艺的基础上进行固态发酵工艺的研究。通过发酵条件和培养基成分的优化,确定了嗜麦芽寡养单胞菌固态发酵羽毛的工艺为:初始pH为7.5,培养温度为35℃,培养时间为4 d,培养基组成是麸皮和干酪素,固液比是1∶1.5,照此工艺发酵,发酵液中蛋白质含量为23.78 mg/mL,角蛋白酶活为20.46 U/mL,菌株的产酶能力及降解羽毛能力显著高于液态发酵条件。     

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。     

N-乙烯基吡咯烷酮降解菌的筛选、降解特性及其竹炭固定化

从活性污泥中分离筛选得到1株N-乙烯基吡咯烷酮(NVP)高效降解菌株ZF1,根据菌株ZF1的形态特征、生理生化特性和16S rRNA基因序列同源性分析,将其初步鉴定为产脲节杆菌(Arthrobacter ureafaciens)。菌株ZF1能以NVP为唯一碳、氮源进行生长,并在60 h内完全降解200 mg·L~(-1)的NVP。菌株ZF1降解NVP的最适环境条件为温度30℃,初始pH 7.0,Na Cl浓度7 g·L~(-1)。为了更好地实现其在实际废水中的应用,将菌株ZF1固定到竹炭上,扫描电镜观察表明ZF1能很好的附着在竹炭表面,且固定菌对NVP的降解效率明显高于游离菌。游离菌和固定菌对NVP的降解均符合一级动力学模型。重复利用5次后,固定菌对NVP的降解率仍能达到98%。     

生物表面活性剂合成条件的优化及提取方法

利用油平板筛选法和排油圈法从辽河某油田的土壤样品中分离筛选高效的生物表面活性剂产生菌,通过16S rRNA基因序列分析对所筛选菌株进行鉴定。采用L16(45)正交实验对所筛选菌株合成生物表面活性剂的条件进行优化。比较5种提取方法对发酵液中生物表面活性剂的提取效果。共分离得到17株菌,其中菌株A3、As和Y产表面活性剂的能力最强。16S rRNA基因序列分析表明,这3株菌均为假单胞菌属(Pseudomonas)。菌株A3、As、Y和A3-As混合菌的生物表面活性剂提取量较优化前有明显的提高,其产量分别是优化前的8.4倍、6.4倍、5.6倍和5.7倍,其中,菌株As的生物表面活性剂产量最高,可达20.55 g/L。对于3种纯菌和A3-As混合菌而言,5种提取方法中均以CHCl_3萃取的量最高;而对其他混合菌,5种提取方法的差别并不显著。菌株A3、As、Y均属于高效的生物表面活性剂产生菌,具有很好的应用前景。     

海洋芘降解菌Alteromonas sp.P7的降解特性及基因组初步分析

以芘(PYR)为唯一碳源,在青岛受石油烃污染的码头的海底沉积物中分离到1株PYR海洋降解菌株P7,经过形态学观察、16S rRNA基因序列分析鉴定菌株P7属于交替单胞菌(Alteromonas)。该菌株在20 d内对PYR的降解率可达44.8%。经GC-MS鉴定分析发现,菌株P7降解芘的代谢产物中含有邻苯二甲酸和水杨酸,且细胞代谢过程中产生邻苯二酚2,3-双加氧酶,由此推测出Alteromonas sp. P7对芘的降解中间产物中含有邻苯二酚,且对芘的降解是通过邻苯二甲酸途径和水杨酸途径实现的。通过对分离得到的Alteromonas sp. P7进行测序,得到1条全长4 597 467 bp的基因组,对该基因序列注释,得到编码基因8 164个,平均GC含量为44.26%。重将测序结果与数据库比对并检索,菌株P7含有参与编码邻苯二酚2,3-双加氧酶的基因。     

一株降解纤维素梭形芽胞杆菌的筛选与鉴定

从肥沃的土壤中经过分离获得14株降解纤维素的细菌,以菌株在纤维素刚果红平板中溶菌圈直径与菌落直径的比值为依据进行筛选,获得一株比值为4.5的菌株,编号为X62。采用DNS法对影响该菌产生纤维素酶活力的单因素进行了分析,并通过对其进行形态特征、生理生化特征测定以及16S rDNA序列分析,确定X62为梭形芽胞杆菌（Lysinibacillus fusiformis）,将其命名为Lysinibacillus fusiformis X62。X62的16S rRNA序列GenBank登录号为JX198550。     

具有降解亚硝酸盐活性的解淀粉芽孢杆菌的分离与安全性分析

从养殖污泥中分离筛选了一株优良的亚硝酸盐降解菌YX01,经ARIS 2X细菌自动鉴定系统以及16SrRNA系统发育分析,该菌株被鉴定为解淀粉芽孢杆菌(Bacillus am yloliquefaciens,GenBank登录号为JX649949),其16S rRNA序列与基因库中 芽孢杆菌属菌株的16S rRNA序列有99％～100％的同源性,而且与解淀粉芽孢杆菌菌株LD5(GenBank登录号:GQ853414)的亲缘关系最近.该亚硝酸盐降解菌YX01在浓度为2.0×107～2.0×109cfu/L时对小球藻(Chlorella sp.)生长有促进作用;在浓度大于2.0×1011cfu/L时对小球藻生长具有显著抑制作用;亚硝酸盐降解菌YX01对小球藻的半数抑制浓度(IC50)大于2.0×1011cfu/L.亚硝酸盐降解菌YX01对大型蚤(Daphnia magna)的48h-半数致死浓度(LC50)大于2.0×1011cfu/L,对草鱼(Ctenopharyngodon idellus)的96h-LC50大于2.0×1011cfu/kg.经分析,亚硝酸盐降解菌YX01为无毒菌株.     

Effects of Mn(II) and Fe(II) on microbial removal of arsenic (III)

Goal, scope, and background  Arsenic contamination in groundwater creates severe health problems in the world. There are many physiochemical and biological methods available for remediation of arsenic from groundwater. Among them, microbial remediation could be taken as one of the least expensive methods, though it takes longer treatment time. The main objective of this research was to study the improvement on remediation by addition of some essential ion salts such as Mn and Fe. Materials and methods   Staphylococcus aureus, Bacillus subtilis, Klebsiella oxytoca, and Escherichia coli were taken as model microbes from Dhulikhel, 30 km east from Kathmandu, Nepal. Results and discussion  Microbes used in this study showed different abilities in their removal of As(III) with and without addition of Mn and Fe salts. The trend of remediation increased with time. S. aureus was found to be the best among the microbes used. It showed almost 100% removal after 48-h culture, both with and without Fe and Mn salts. Rate of removal of As increased with addition of Fe and Mn for all microbes. Removal efficiency was found to increase by about 32% on average after addition of salts in 24-h cultures of S. aureus.     

Patterns of volatile metabolites and nonvolatile trichothecenes produced by isolates of <Emphasis Type="Italic">Stachybotrys,Fusarium, Trichoderma,Trichothecium</Emphasis> and <Emphasis Type="Italic">Memnoniella</Emphasis>

We reported previously that trichodiene, a volatile trichothecene derivative, was produced by a Stachybotrys isolate, also known to produce highly cytotoxic, non-volatile, macrocyclic trichothecenes (satrotoxins). We investigated the relationship between the production of trichodiene and various non-volatile trichothecenes for several molds. Volatile metabolites were concentrated by adsorption on Tenax TA and analyzed by GC/MS, while non-volatile metabolites were separated by HPLC, derivatized and analyzed by GC/MS. Stachybotrys chartarum isolates producing macrocyclic trichothecenes secreted significantly larger amounts of trichodiene and other sesquiterpenes than isolates which only produced simple trichothecenes. The amounts of secreted trichodiene were relatively small in all cases. With the exception of Memnoniella, which excreted small amounts of sesquiterpenes, the other isolates produced varying amounts of sesquiterpenes, including trichodiene, as well as simple tricothecenes, no detectable trichodiene, but large amounts of griseofulvin derivatives. In Stachybotrys there is apparently a correlation between trichodiene and macrocyclic trichothecene production. In the remaining isolates, there was no simple relationship between trichodiene and non-volatile trichothecene synthesis. Trichodiene is produced in larger amounts by Stachybotrys isolates, which also produce satratoxins, but it will be difficult to utilize this metabolite to detect toxic isolates in buildings due to the relatively small amounts excreted.     

Bioindicators for toxicity assesment of effluents from a wastewater treatment plant

The discharge of effluents and toxic compounds into aquatic systems represents a growing environmental problem involving an impact on water ecology and potential effect on human health. Most municipal wastewaters are complex mixtures; their complexity led as to carry out a hazard assessment using chemical analyses and biological tests.This study investigates biochemical alteratiobns in two sentinel organisms, the Anodonta cygnea mussel and the Xenopus laevis frog exposed for different lengths of time to various concentrartion of wastewater of the S. Antonino Ticino treatment plant. The results point out the long-life of toxic compounds.     

利用发光菌和大型蚤对北方某城市再生水急性毒性的调查

利用发光菌和大型蚤作为受试生物测定了北方某城市5个再生水原水(城市污水厂二级出水)和2个再生水处理系统的各工艺出水的急性毒性.结果表明,各再生水原水对大型蚤和发光菌具有不同程度的毒性效应,其中工业废水占较大比例的K和B厂再生水原水的大型蚤48 h总抑制率分别高达90%和100%,发光菌发光抑制率分别达到74.2%和46...     

氧化亚铁硫杆菌浸铜作用研究

为研究氧化亚铁硫杆菌(Acidithiobacillus ferrooxidans,简称A.f)对铜浸出的作用,从某煤堆积水中分离得到A.f菌,利用该菌对铜进行浸出实验,设置3个处理,3个处理的浸出液分别为9 K培养基(简称S),成熟菌液(简称MS)和过滤除菌液(简称FS)。结果表明,3个处理中铜都得到了浸出,浸出铜浓度按S、FS和MS依次递增分别为4 433、5 377和6 296 mg/L;浸出初期,MS中的铜,浸出速度最快,过滤除菌液中次之,而培养基中的速度比较稳定,比前两者均慢,24 h后三者浸铜速度趋近一致;三者中pH、Eh变化相似,pH均先升高至3.4左右保持相对稳定,Eh均先迅速下降至280 mV后保持稳定。由实验可知,氧化亚铁硫杆菌主要在浸出初期促进了铜的浸出,且菌液中溶解氧氧化Fe2+以及酸性条件下氧化Cu0对铜的浸出作用不容忽视。     

污泥堆肥施用于草坪草栽培:Zn和Cu的分布特征简

为了探索污泥堆肥中重金属在土壤-植物系统中的积累与转移特性,通过温室盆栽实验,分析了污泥堆肥对草坪草高羊茅、黑麦草和白三叶生物量积累的情况,研究了污泥堆肥中Zn和Cu在植物和土壤中的分布特征。结果表明,污泥堆肥施用可以有效促进3种草坪草的积累生物量,在0~6 kg/m²的污泥堆肥施用量范围内,草坪草的生物量积累随着施用量的增加而提高。土壤中Zn和Cu的含量随污泥堆肥施加量的增加而增大,85%以上的Zn和Cu残留在土壤中。污泥堆肥中的Zn和Cu均可以被植物吸收,随着污泥堆肥施用量的增加,草坪草对Zn和Cu的吸收量增大,但当污泥堆肥施用量超过一定阈值时,草坪草吸收Zn和Cu不再增加,甚至减少;对于不同的草坪草,这一阈值有所不同。植物对Zn和Cu的吸收量只占土壤中Zn和Cu减少量的5%左右。根据生物富集系数（BCF）的计算结果推测,污泥堆肥的施用对土壤环境的影响大于对植物体内累积Zn和Cu的影响。     

Marine macroalgal community structure, metal content and reproductive function near an acid mine drainage outflow

Marine macroalgal communities were examined near the outflow of acid mine drainage (AMD) from the Britannia Mine, British Columbia, Canada. No marine algae were present within 100 m of the mouth of Britannia Creek, which carries the AMD into the marine environment. At greater distances (300-700 m) from this Creek, mean summer cover of filamentous green algae, mostly Enteromorpha intestinalis, was >60%, which was significantly higher than at nearby reference stations. At still greater distances (600-1000 m) from Britannia Creek, Fucus gardneri dominated algal communities that were similar to those at reference stations. No consistent differences were detected in mean plant length, mean per cent cover or mean oocyte production between F. gardneri near Britannia Creek and those at reference stations. Cu body burden in F. gardneri near Britannia Creek was five to 17 times higher than in reference plants.     

不同质量浓度黄菖蒲和狭叶香蒲对铜绿微囊藻的化感作用简

分别采用黄菖蒲（Iris pseudacorus L.）、狭叶香蒲（Typha angustifolia L.）和铜绿微囊藻（Microcystis aeruginosa）共生培养的实验方法研究了不同质量浓度黄菖蒲、狭叶香蒲对铜绿微囊藻的化感作用。结果表明,黄菖蒲在质量浓度大于10 g/L时对初始密度为1.0×10⁷ ind/mL的铜绿微囊藻具有较好的抑制作用,表现为黄菖蒲质量浓度为10、20和40 g/L时,第15天对铜绿微囊藻的抑制率分别为30.1%、51.8%和84.0%;狭叶香蒲在质量浓度大于20 g/L时对铜绿微囊藻有明显的抑制作用,表现为狭叶香蒲质量浓度为20 g/L和40 g/L时,第15天对铜绿微囊藻的抑制率分别为34.2%和77.7%,实验过程中,铜绿微囊藻叶绿素a含量逐渐减少,而藻密度、SOD活性及MDA含量先增加后逐渐降低,表明经过一段时间持续地化感胁迫,黄菖蒲和狭叶香蒲可以诱导铜绿微囊藻产生氧化胁迫,导致细胞结构严重损伤和叶绿素大量分解,从而强烈抑制铜绿微囊藻的生长。     

Physical-chemical and ecotoxicological evaluation of water based drilling fluids used in Italian off-shore

In order to evaluate the effects on the marine ecosystem caused by an eventual discharge into sea of water based drilling fluids, as current legislation allows, chemical and ecotoxicological analyses were performed on the most common drilling muds and products used in Italian off-shore activities. The chemical analysis on drilling fluids involved the leaching test and the measurement of total content of heavy metals, whereas biodegradation tests were performed on the products used in mud's formulations. As for ecotoxicological evaluation, two marine organisms, the crustacean Artemia salina and the diatom Phaeodactylum tricornutum, were selected to determine the LC₅₀ and the EC₅₀ respectively.