太湖土著田螺消化道中溶藻菌XMC溶藻进程与叶绿素a降解动力学研究

在蓝藻爆发区域的太湖土著田螺消化道内,筛选出l株高效溶藻菌,命名为XMC.以铜绿微囊藻为受试对象,以叶绿素a含量检验溶藻菌XMC溶藻效果,考察了菌株溶藻能力、溶藻过程及其溶藻产物等.实验结果表明,溶藻菌XMC具有较强的溶藻能力,其生长曲线呈"S"型增长,符合Logistic动力学模型.菌藻共生环境中,藻液叶绿素a含量与...     

干粉菌剂保护剂对溶藻菌溶藻效果与稳定性能的影响

针对液体菌剂在使用过程中易受杂菌污染以及运输过程中存储不便等问题,选取蔗糖、甘油、谷氨酸钠、脱脂乳粉等4种常见材料做干粉菌剂保护剂,真空冷冻干燥条件下,以小麦粉作为载体,分析不同浓度保护剂作用下的活菌数量及冷冻干燥前后的活菌率变化,作为探究4种保护剂在干粉菌剂制备过程中保护作用与机理的依据.实验结果表明,浓度为10%蔗糖溶液、10%甘油溶液、4%脱脂乳粉溶液以及8%谷氨酸钠溶液做保护剂时活菌率最高、活菌数最大,蔗糖浓度为10%,冷冻干燥前液体菌剂中溶藻菌的数量约为9.65×10~9 CFU·mL~(-1),冷冻干燥后干粉菌剂中溶藻菌数量约为15.20×10~9 CFU·mL~(-1),溶藻菌成活率为157.51%,高于其他3种保护剂.整个溶藻过程大致持续8 d,在第8 d时,蔗糖作保护剂的菌剂溶藻率达96.04%,以甘油作保护剂菌剂溶藻率达96.11%,菌剂中含有谷氨酸钠和脱脂乳粉,溶藻率分别为94.14%和94.22%.蔗糖和甘油在菌体新陈代谢过程中对菌的保护能力较强,溶藻菌菌体数量多,成活率高,菌体稳定性好,溶藻效果比液体菌剂稍差.     

一株溶藻细菌的分离鉴定及溶藻效果

从太湖分离到一株溶藻细菌CA,该菌对铜绿微囊藻具有强烈的溶藻效果.通过形态学、生理生化特征及16SrDNA序列比对,鉴定该菌株属于水单胞菌属(Aquimonas sp.).将CA菌悬液与铜绿微囊藻共培养,10 d内铜绿微囊藻细胞降解率为100%,叶绿素a降解率为83.9%,且溶藻效果与菌悬液浓度呈正相关,与藻浓度呈负相关.CA菌体本身没有溶藻效果,但其无菌滤液可以溶藻,因此CA是通过释放物质来间接溶藻.在CA菌体的菌藻共培养液中添加少量的牛肉膏、葡萄糖或尿素,菌体显示出溶藻效果.本研究为菌株CA控制铜绿微囊藻水华提供了一种潜在的应用前景.     

溶藻细菌胶囊与植物共生系统的建立及其对Microcystis aeruginosa的控制

当前,蓝藻水华时常爆发,其中铜绿微囊藻(Microcystis aeruginosa)是我国蓝藻水华的典型代表.微生物控藻技术具有高效、生态安全性好、原位修复等特点,近年来已经成为治理蓝藻水华的主要手段.本文通过搭建芦苇湿地试验装置,模拟太湖近岸水域原生环境,以水生植物控藻与溶藻细菌胶囊强化相结合的方式对Microcystis aeruginosa进行生态学-微生物操控试验.从藻胆蛋白、抗氧化系统、膜脂值活性等方面的研究来探究控藻系统的溶藻机制,并通过高通量技术与主成分分析来探索溶藻细菌与藻之间的群落动态关系.试验结果表明,原位藻类控制系统效率高,14 d溶藻率为(88.74%±1.10%),其中溶藻细菌胶囊占主导地位,植物修复为辅.通过对藻胆蛋白的破坏和藻类抗氧化系统的摧毁来导致藻类膜结构的受损,胞内物质流出而死亡.利用高通量技术发现,溶藻细菌胶囊所包埋的菌株Bacillus sp.HL在微生物体系中成为优势菌种.主成分分析结果表明高效原位控藻体系促进了藻体内ROS水平,增强了藻的Zeta电位和抗氧化酶活性,从而抑制了Microcystis aeruginosa的生长.     

一株溶藻细菌对铜绿微囊藻的溶藻机理初探

为确定溶藻细菌S7（Chryseobaterium）对铜绿微囊藻的溶藻方式,分别采用高温灭菌（121~123℃）、离心（10 000 r.min-1）、0.22μm滤膜过滤等方式对S7菌液进行处理,检测其对铜绿微囊藻的去除效果。并通过对溶藻过程中叶绿素a和丙二醛（MDA）含量的测定,藻细胞显微结构的观察和细胞成分的红外光谱分析,初步探讨菌株S7对铜绿微囊藻的作用机理。结果表明,S7是通过释放胞外活性物质间接溶藻,该物质具有很强的热稳定性,不属于蛋白质类物质。该活性物质对铜绿微囊藻的叶绿素a有明显的去除效果,并可导致藻细胞膜脂过氧化产物MDA积累量的显著提高和藻细胞解体。藻细胞红外光谱分析表明,经过溶藻物质作用的藻细胞,其蛋白质结构遭到破坏。通过试验结果,推测出菌株S7的溶藻机理：溶藻物质先损伤铜绿微囊藻的细胞壁和粘质胶被,然后通过改变膜的选择透过性进入藻细胞内部,分解叶绿素a,破坏蛋白质,造成藻体正常生理功能的丧失,最终导致藻细胞破裂。     

三株溶藻细菌溶藻活性代谢产物的初步研究

已知3株溶藻细菌L7、L8和L18的活性代谢产物具有明显的溶藻效果。为获得较高活性和浓度的目标产物,研究了培养基、碳源、氮源对溶藻效果的影响;为考察溶藻活性代谢产物保存和应用的环境条件,研究了其热、酸稳定性。在牛肉膏蛋白胨、淀粉、查氏和高氏1号4种培养基里,淀粉培养基最适宜用于获得溶藻活性代谢产物。以淀粉培养基为基础,碳、氮源组合依次为淀粉 (NH4)2SO4、淀粉 (NH4)2SO4,葡萄糖 KNO3时,3株溶藻细菌的溶藻活性代谢产物溶藻活性最高。这一结果为目标产物的分离奠定了物质背景。3株溶藻细菌溶藻活性代谢产物均具有良好的热稳定性,经热处理后,对叶绿素a的去除率仍高于73%。L7的无菌滤液调至pH值4.0或2.5,2h后丧失溶藻活性;L8和L18的无菌滤液调至pH值4.0,2h后未丧失溶藻活性,调至pH值2.5,2h后丧失溶藻活性。上述结果为溶藻活性代谢产物的分离奠定了基础。     

溶藻细菌DC-L14的分离、鉴定与溶藻特性

从滇池蓝藻水华集聚区分离获得一株溶藻细菌DC-L14,经16S rDNA序列分析鉴定为Lysinibacillus fusiformis;小白鼠毒性试验初步显示该菌株未产生小白鼠中毒毒素;该菌能使铜锈微囊藻905聚集成团,沉于瓶底,最终黄化;该菌作用4 d,使惠氏微囊藻107、绿色微囊藻102、水华束丝藻和水华鱼腥藻的叶绿素a下降率最高为70.1%.最低为65.5%,平均为67.2%;当细菌处于稳定生长期时溶藻效果最强,共培养4 d能使铜锈微囊藻905的叶绿素a含量下降82.1%;离心沉降后检测,发现菌体本身无溶藻效果,而无菌上清液与原菌液溶藻效果相同,高温处理后的菌液溶藻能力增强,推测该细菌是通过分泌溶藻物质溶藻,该物质可能为非蛋白质类,高温可能有利于溶藻物质的释放.图4表1参25     

溶藻细菌及其分子生物学研究进展

溶藻细菌在"水华"防治中的作用和潜力,已广受关注.本文从系统分类、溶藻机制和杀藻物质等方面对已报道的溶藻细菌的研究结果进行了归纳和总结,并对近年来在细菌溶藻研究中应用的分子生物学技术,包括PCR、核酸探针和全细胞杂交、变性梯度凝胶电泳(DGGE)等,以及溶藻机制研究巾的分子生物学进展进行了综述,最后提出了在溶藻细菌的研究和应用中值得注意的几个问题.表1参66     

溶藻菌对受污染水源水除藻及脱氮特性研究

针对我国水源地藻类污染日趋严重等问题,利用前期分离获得的溶藻菌Streptomyces sp.HJC-D1研究固定化微生物技术强化污染水源水除藻以及脱氮性能。结果表明,对照组和试验组的水体叶绿素a平均去除率分别为（71.66±5.35）%和（80.94±4.36）%,NH4＋—N的平均去除率为（77.76±2.83）%和（72.36±3.18）%,而高锰酸盐指数（CODMn）平均去除率为（24.99±1.52）%和（18.74±1.38）%;不同曝气条件的影响研究发现,曝气/停曝时间比2：4、曝气量60 L.h-1工况下,系统CODMn和NH4＋—N去除率均有所提高,相比对照组NO3-—N积累更为明显;水力停留时间（HRT）变化对系统NH4＋—N、CODMn等的去除影响不大,但缩短HRT时叶绿素a去除率有所降低;分析反应器内填料表面微生物相发现,试验组填料表面有溶藻菌富集,推测对照组除藻主要通过填料对藻类的吸附去除,而试验组则是藻类吸附在填料表面后通过溶藻微生物实现藻类去除。     

溶藻放线菌对铜绿微囊藻和小球藻竞争生长的影响

从土壤中分离获得l株对铜绿微囊藻有明显抑制作用的橄榄网状链霉菌SG-001（Streptomyces olivoreticuli SG-001）,研究其对铜绿微囊藻（Microcystis aeruginosa）和小球藻（Chlorefla pyrenoidosa）竞争生长的影响。结果表明：SG-001菌株的活性物质主要存在于无菌滤液中,能够强烈抑制铜绿微囊藻的生长,但对小球藻的生长具有明显的促进作用。当混藻中两种藻细胞初始接种浓度均为4.0×106 mL-1时,在BG11纯培养条件下,添加SG-001无菌滤液有利于小球藻生长,但对铜绿微囊藻抑制作用不明显;而SG-001无菌滤液对天然加富水样中的铜绿微囊藻具有明显抑制作用,在同时接种有铜绿微囊藻和小球藻的混合藻液中,添加SG-001无菌滤液能够明显提高小球藻的生长竞争能力,且水体中氨氮和可溶性总磷的去除率可分别达到85%和93.33%,而铜绿微囊藻在第8天时生长基本被小球藻抑制。     

纳米银和银离子对2种微藻的急性毒性效应

为了探讨AgNPs对典型微藻的急性毒性效应及其机制,采用柠檬酸钠还原法制备AgNPs,以摇瓶实验法评估了不同浓度的AgNPs和Ag+对铜绿微囊藻和普通小球藻叶绿素a含量、形态结构和叶绿素荧光参数的影响。实验结果表明:AgNPs对普通小球藻和铜绿微囊藻的96 h-EC50分别为1.113 mg·L-1和0.697 mg·L-1,而Ag+对2种藻的96 h-EC50分别为0.106 mg·L-1和0.032 mg·L-1。扫描电镜结果表明:AgNPs处理使普通小球藻细胞表面出现褶皱,细胞变形甚至向内塌陷。对铜绿微囊藻部分细胞出现变形变得不规则,且出现某些胞外物质使细胞粘附在一起。透射电镜观察发现,高浓度Ag+处理使2种藻的细胞均发生质壁分离,部分细胞转变为孢子。而AgNPs处理使普通小球藻细胞蛋白核增大,蛋白核与类囊体区无明显连接通道。铜绿微囊藻拟核区膨大,类囊体和色素体被推向四周,部分类囊体断裂,同时,发现该藻可以分泌胞外物质在细胞周围吸附AgNPs颗粒。对于普通小球藻,0.6 mg·L-1AgNPs处理后细胞光系统Ⅱ的最大光化学量子产率ΦP0相对于CK没有显著差异,但0.09 mg·L-1Ag+处理使ΦP0显著增加。在高浓度AgNPs或Ag+处理时,ΦP0均显著降低。AgNPs未对普通小球藻光系统II性能参数PI_Abs造成影响,但不同浓度Ag+处理均使得该参数显著升高。对于铜绿微囊藻,2种毒物均使其ΦP0显著降低。而PI_Abs仅在2种毒物的最高浓度处理时显著降低。综上,AgNPs对2种藻的急性毒性远小于Ag+,而两者对铜绿微囊藻的毒性均大于普通小球藻。AgNPs胁迫使2种藻叶绿素a含量显著降低,并诱导2种藻在形态结构和光合生理方面发生了显著变化,造成不同程度的损伤,但与Ag+的毒性效应存在一定的差异。提高光吸收能通量补偿耗散能量和分泌胞外物质结合Ag+是微藻2种重要的解毒机制。     

光合细菌对铜绿微囊藻生长的抑制效应

为了探讨光合细菌对水华藻类的控制作用,在实验室条件下,通过菌藻共同培养,研究了沼泽红假单孢菌(Rhodopseudomonas palustras)、球形红细菌(Rhodobacter sphaeroides)及其混合培养物对铜绿微囊藻(Microcystis aeruginosa)生长的抑制效应.实验结果表明,光合细菌混合培养物对铜绿微囊藻抑藻作用最强,培养5 d时,铜绿微囊藻生物量降低率达58.9%,培养时间、培养温度、菌体投加量及培养基pH值等对光合细菌混合培养物的抑藻作用均有不同程度的影响.光合细菌混合培养物发挥抑藻作用的适宜条件为培养温度25℃,培养时间≤5 d,光合细菌投加量(V_(光合细菌菌悬液):V_(藻培养液)=1:80),藻培养基pH为8.0左右.     

典型抗生素二元混合物对明亮发光杆菌的急性联合毒性

以明亮发光杆菌作为受试生物,测定了不同类型抗生素的二元混合物对明亮发光杆菌的等比急性联合毒性.结果表明,4类抗生素对明亮发光杆菌二元等比急性联合毒性主要呈加和效应及拮抗效应.本研究可为水体中抗生素的联合生态风险评价和修复提供理论依据.     

Acute toxicity of roundup to the nontarget organism Hyalella curvispina. Laboratory and field study

Glyphosate is the most used pesticide in Argentina. Hyalella curvispina is a widely distributed and commonly abundant component of the invertebrate assemblages in shallow waters of southern South America. The aim of this study was to assess the acute toxicity of the increasingly common Roundup Full II®, commercial formulation of the herbicide glyphosate (66.2% active ingredient), to H. curvispina in laboratory and field assessments. The mean estimated 48-h LC₅₀ of Roundup was 9.9 ± 1.7 mg L^?1. In a field experiment Roundup was applied to soybean plots. Simulated rain was generated the following day by means of irrigation sprinkler equipment. H. curvispina was exposed to runoff water and soy leaves. No mortality was observed. It is suggested that Roundup crop applications represent a low risk of acute toxicity to H. curvispina adults inhabiting water bodies adjacent to crop fields.     

等毒性配比法研究镉、铬和铅对淡水发光细菌的联合毒性

当待测生物暴露在混合污染物中时,由于混合物中各组分相互影响,会产生联合毒性作用,表现为加和作用、协同作用和拮抗作用。为了深入了解重金属混合物的联合毒性对发光细菌的作用,利用淡水发光细菌——青海弧菌Q67(Vibrio qinghaiensis sp.nov-Q67)发光值的测定方法,采用联合毒性单位法,在测定了硝酸镉、重铬酸钾和硝酸铅单一毒性EC50的基础上,对硝酸镉 重铬酸钾、硝酸镉 硝酸铅、硝酸铅 重铬酸钾3种重金属二元混合物的联合毒性进行了评价。结果表明,硝酸镉 重铬酸钾、硝酸铅 重铬酸钾是拮抗作用,硝酸铅 硝酸镉是协同作用。     

Cadmium and lead toxicity and bioaccumulation in Microcystis aeruginosa

The growth of human population leads to intensification of agriculture and promotes, through eutrophication, development of cyanobacteria. One of the most widespread and bloom-forming species in freshwater is toxic Microcystis aeruginosa （M. aeruginosa）. Combustion of fossil fuels and metallurgical processes are the main sources of heavy metals contamination in surface water including cadmium （Cd） and lead （Pb）. The following study was conducted in order to determine the effect of 1- 20 mg. L-1 of Cd and Pb on photochemistry （using flow cytometry） and growth （based on chlorophyll concentra- tion） ofM. aeruginosa as well as to estimate levels of metal bioaccumulation. We have found that 1-10mg.L-1 of Cd and 1-5 rag. L1 of Pb induced continuous enhancement of chlorophyll fluorescence during 24 h of incubation. No significant degradation of chlorophyll was observed in these samples. At higher concentrations of 20 mg. L-1 of Cd and 10-20 mg.L-1 of Pb chlorophyll level significantly decreased and its fluorescence was quenched. M. aeruginosa demonstrated high capability of Cd and Pb bioaccumulation, proportionally to initial metal concentration. In samples with initial concentration of 20 mg. L-1 of Cd and Pb bioaccumulation of 87.3% and 90.1% was observed, respectively. Our study demonstrates that M. aeruginosa can potentially survive in highly metals polluted environments, be a primary source of toxic metals in the food chain and consequently contribute to enhanced toxicity of heavy metals to living organisms including human.     

Acute toxicity assessment of drinking water source with luminescent bacteria: Impact of environmental conditions and a case study in Luoma Lake,East China

• Acute toxicity assessment was conducted in Luoma lake watershed, East China. • Impacts of environmental factors on the toxicity testing was fully evaluated. • Dissolve oxygen had a weak positive correlation with luminescence inhibition rate. Protecting the quality of lake watersheds by preventing and reducing their contamination is an effective approach to ensure the sustainability of the drinking water supply. In this study, acute toxicity assessment was conducted on the basis of acute bioluminescence inhibition assay using the marine bacterium Vibrio fischeri as the test organism and Luoma Lake drinking water source in East China as the research target. The suitable ranges of environmental factors, including pH value, organic matter, turbidity, hardness, and dissolved oxygen of water samples were evaluated for the toxicity testing of bioluminescent bacteria. The physicochemical characteristics of water samples at the selected 43 sites of Luoma Lake watershed were measured. Results showed that the variations in pH value (7.31–8.41), hardness (5–20 °d) and dissolved oxygen (4.44–11.03 mg/L) of Luoma Lake and its main inflow and outflow rivers had negligible impacts on the acute toxicity testing of V. fischeri. The luminescence inhibition rates ranged from -11.21% to 10.80% at the 43 sites. Pearson’s correlation analysis in the experiment revealed that temperature, pH value, hardness, and turbidity had no correlation with luminescence inhibition rate, whereas dissolved oxygen showed a weak statistically positive correlation with a Pearson correlation coefficient of 0.455 (p<0.05).     

Acute toxicity of benzophenone-type UV filters and paraben preservatives to freshwater planarian,Dugesia japonica

Fourteen benzophenone-type UV filters and four paraben preservatives were selected to examine their acute toxicities on Dugesia japonica. The 48-h LC₅₀ values for planarians exposed to benzophenone-type UV filters can be ranked as oxybenzone?>?mexenone?>?5-chloro-2-hydroxybenzophenone?> 2,4-dihydroxybenzophenone?>?2-hydroxybenzophenone?>?dioxybenzone?>?benzophenone?>?2,2′,4,4′-tetrahydroxybenzophenone?>?4-hydroxybenzophenone?> 3-hydroxybenzophenone?>?4,4′-dihydroxybenzophenone?>?2,2′-dihydroxy-4,4′-dimethoxybenzophenone?>?2,3,4-trihydroxybenzophenone?>?sulisobenzone with a range from 0.9 to 145?mg?L^?1 with a similar sequence for the 96?h LC₅₀ values, ranging from 0.5 to 77?mg?L^?1. The 48 and 96?LC₅₀ values for planarians exposed to paraben preservatives can be ranked as butylparaben?>?propylparaben?>?ethylparaben?>?methylparaben. Among all the tested chemicals, oxybenzone was the most toxic and sulisobenzone the least toxic chemical to planarian at each exposure period. Most benzophenone-type UV filters are toxic to aquatic animals with 48?h LC₅₀ values less than 10 mg?L^?1, except for 2,2′-dihydroxy-4,4′-dimethoxybenzophenone, 2,3,4-trihydroxybenzophenone, and sulisobenzone. Because of their common occurrence in aquatic environment, more studies on aquatic toxicities of benzophenone-type UV filters and paraben preservativs are needed to provide important information to adequately assess their ecological risk.     

Screening for acute ocular toxicity of commercialized products of cartap,an insecticide,and correlation to blood concentrations in rabbits

Cartap is an insecticide known to produce contraction and damage to mouse and rabbit diaphragms, and plays a pivotal role in respiratory paralysis and mortality in rabbits. The aim of this study was conducted to screen for the toxic properties and blood concentrations of commercialized cartap products in New Zealand White (NZW) rabbits. Five granular formulations (GRs) with different concentrations (4, 6, 6.5, 6.66 or 10%) and seven water-soluble powders (SPs, 50%) of commercialized cartap products were chosen to study toxicity following ocular instillation. Furthermore, ocular instillation (5, 12.5, 25 or 50 mg kg^?1) and oral administration (25 mg kg^?1) of the technical material (TC) cartap was also performed to compare blood concentration changes in rabbits. There was no death or eye irritation with all GR products; however, soluble powder (SP) products produced severe toxicity and mortality in rabbits. The blood concentration of TC cartap in rabbits started to increase from 2 min and reached peak levels at 7–9 min after ocular instillation in a dose-related pattern. For comparison of the blood distribution of cartap via different administration routes, oral exposure exhibited a higher absorption of cartap than ocular instillation. It was suggested that the commercialized GR products of cartap were safe but 50% SP products were highly toxic to rabbits following ocular exposure. This study provides valuable data to screen for potential acute toxicity of cartap to farmers and animals in the field.     

载钯树脂对偶氮类染料的吸附降解及其产物对发光菌的毒性暴露试验

合成了表面负载金属钯的聚丙烯醛异烟酰腙树脂后,通过Q67发光菌的毒性暴露实验研究了该树脂材料对直接大红、直接天蓝和桃红等3种水溶性偶氮染料的催化降解效果及其在降解过程中对发光菌的生物毒性变化情况,并对降解结果进行了气相色谱-质谱(GC-MS)分析.实验结果表明:载钯树脂对直接大红、直接天蓝和桃红等偶氮染料具有良好的吸附...