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.     

Pb2+、Cd2+和Cr(Ⅵ)抑制球形红细菌生长的毒性效应

运用评价化学品对水生生物毒性的标准方法,得到Pb2 、Cd2 和Cr(Ⅵ)对球形红细菌生长的最小无差异浓度(LNOEC)分别为85.9mgL-1、22.1mgL-1和17.7mgL-1.根据实验结果采用几率单位进行数据处理,得到抑制球形红细菌生长的96hEC50分别为195.0mgL-1、91.2mgL-1和77.6mgL-1,并对各剂量反应方程进行χ2检验,结果均符合精度要求,计算出的96hEC50真实可靠.研究结果表明,抑制球形红细菌生长的毒性由大到小的顺序是Cr(Ⅵ)>Cd2 >Pb2 .不同金属离子与细胞壁亲和性不同或能通过菌体内生物化学转化形成沉淀,可能是导致金属离子抑制球形红细菌生长毒性差异的主要原因.图1表2参15     

假单胞茵胞内酶粗提液对藻毒素MCLR的降解

对比研究了假单胞菌M-6及其细胞内外提取液对微囊藻毒素-LR(MCLR)的降解效率.结果表明,细胞外提取液对藻毒素没有降解作用,胞内酶粗提液能在24h内降解MCLR,日均MCLR降解率是纯菌株M-6的4.7倍.进而研究了酶蛋白浓度、底物MCLR浓度、pH值及环境温度对胞内酶粗提液降解藻毒素效率的影响.当MCLR浓度为15.0mg·l~(-1)时,MCLR适宜的酶促降解反应条件为:酶蛋白浓度280mg·l~(-1),温度为30℃,反应pH值为7.0.MCLR液相色谱结构变化图表明,至少有3种酶参与了胞内酶粗提液降解MCLR的分子过程.     

几株紫色非硫细菌的分离鉴定与产氢特性分析

从重庆地区不同环境淤泥、泥水样品中,经富集培养、分离纯化,获得5株紫色非硫细菌.根据菌体的菌落形态、染色特性、生理生化特征及活细胞光吸收峰对菌株进行常规鉴定,结合菌株16S rDNA扩增测序进行分子生物学分析验证,构建了菌株与数据库中近缘菌株的系统发育树.以优化的培养条件(营养、pH、接种量等参数)对供试菌株的生理生化特性和产氢能力做了比较分析.结果显示,5个菌株均为光合产氢细菌,菌株ANI、D1为沼泽红假单胞菌(Rhodopseudomonas palustris),AN2、AS1、BS1等3株为类球红细菌(Rhodobacter sphaeroides);其中类球红细菌菌株AN2在给定的培养条件下光合产氢能力最高,可达9.55μg/mL d-1,是一株有应用前景的光合产氢细菌.图4表2参15     

雪灵芝不同提取物的生物活性

采用蛋白酪氨酸磷酸酯酶(PTP1B)、二肽基肽酶(DPP-IV)、α-葡萄糖苷酶-rat(α-glucosidase-rat)、MCF-7/ADR肿瘤耐药细胞增敏、肿瘤细胞毒活性(HepG2肝癌细胞)、骨髓有核细胞增殖等模型,对雪灵芝水提物、醇提物和市售的雪灵芝营养口服液分别进行体外免疫调节、抗肿瘤、糖尿病等活性筛选.结果表明:雪灵芝水提物、醇提物和营养口服液均对蛋白酪氨酸磷酸酯酶/PTP1B模型呈阳性,对其余5种模型呈阴性,说明蛋白酪氨酸磷酸酯酶(PTP1B)模型可以作为雪灵芝有效部位的活性跟踪模型.     

粘红酵母(Rhodotorula glutinis CIBAS A 1401)苯丙氨酸解氨酶cDNA核心序列的克隆与分析

L苯丙氨酸因其在工业、医药上的重要作用而日益受到人们的重视.苯丙氨酸解氨酶(PAL)生物转化肉桂酸生产L苯丙氨酸成了该领域研究与开发的热点.本文利用简并寡核苷酸PCR法结合cDNA末端快速扩增PCR法,从粘红酵母中克隆了苯丙氨酸解氨酶cDNA核心序列,为克隆其cDNA全长序列及体外表达其活性蛋白奠定了基础.图5参22     

固定CO2氢细菌的筛选及其培养条件优化

从土壤中分离筛选到一株高效固定CO2 的氢氧化细菌—专性化能自养菌BHA 15 ,在最佳培养条件下(培养基 ρ/gL-1:NH4 Cl 4.0 ,Na2 HPO4 ·12H2 O 0 .75 ,KH2 PO4 0 .5 ,MgSO4 ·7H2 O 0 .3;V(H2 )∶V(O2 )∶V(CO2 ) =3 .5∶1∶1;pH =7;最适生长温度θ =30℃ ) ,密闭摇瓶中恒温 (30℃ )培养 40h后 ,菌体浓度 ρ(DW)达 1.6mg/mL ,菌体蛋白含量 71.8% .图 7表 2参 12     

从武汉印染厂废水淤泥中分离出的趋磁细菌的生长与磁小体形成过程

对从武汉印染厂污泥中分离出的趋磁细菌YN-1,分别进行了5 d,10 d,20 d,25 d和30 d的纯培养与透射电镜的观察,并对30 d释放出的磁小体进行了能谱分析.结果表明:培养5 d的趋磁细菌,其体内的细胞质相对均匀,可见到铁质在细胞内输送的痕迹.随后细胞质变为不均匀,菌体内泡状物和磁小体增多;20 d后菌体膜出现自溶,可见磁小体从破裂的菌膜排出体外的现象,25 d后可见到具有鞭毛、细胞质分布均匀的“营养体”.培养30 d,菌膜基本自溶,除见有营养体外,残存大量的细胞膜碎片和磁小体,对它们的能谱分析显示,磁小体的主要无机成分为Fe.由此判断,25 d可能为该菌在培养条件下的趋磁细菌生长与磁小体形成周期.图3表1参15     

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.     

果胶酶激发子对黄瓜叶片病程相关蛋白及细胞壁物质的影响

以草酸青霉菌(Penicillium oxalicum)的固态发酵产物———果胶酶粗提液喷雾诱导黄瓜,研究了该激发子对黄瓜细胞内几丁质酶和-β1,3-葡聚糖酶2种病程相关蛋白(PR s蛋白)以及细胞壁物质的影响.结果表明,经激发子处理后,黄瓜叶片中几丁质酶和-β1,3-葡聚糖酶活性诱导升高,木质素及富含羟脯氨酸糖蛋白(HRGP)含量也诱导增强;同时,在激发子处理后接种黑星病菌的黄瓜中,上述2种PR s蛋白及细胞壁物质被大量诱导合成.其中,两种PR s蛋白均在接种后d 4达到最高值,分别为22.72 U/mg(pro)和268.16 U/mg(pro);木质素和HRGP含量在接种后d 3后达最高,分别比对照高196.12%和272.98%.表明果胶酶激发子在接种前期对黄瓜几丁质酶、-β1,3-葡聚糖酶、木质素及HRGP的诱导产生是导致病情降低的内在机制.图4参16     

Studies on in vitro degradability of mixed crude enzyme extracts produced from Pleurotus spp

A preliminary investigation was conducted to assess lignocellulolytic efficiency of crude extracts from three white-rot fungi, Pleurotus florida PF05 (PF), Pleurotus sajor-caju PS07 (PS) and Pleurotus eryngii PE08 (PE). The activities of CMC-ase, xylanase, beta-glucosidase, beta-xylosidase, laccase and Mn peroxidase in extracts were evaluated. PF produced its highest CMC-ase (317 UL(-1)'), beta-glucosidase (62 UL(-1)), beta-xylosidase (37 UL(-1)) and laccase (347 UL(-1)) activities while, PS produced highest xylanase (269 UL-(1)) and Mn peroxidase (69 UL(-1)) activities. In addition, crude extracts extracted were employed for their in vitro degradability assessment; and were evaluated with mono and mixed extracts separately to corn cob substrate. The losses in cell wall components and dry matter during 5 and 10 days incubations were analyzed after treatments of extracts. Maximum 8.2, 4.4 and 2.8% loss were found respectivelyin hemicellulose (HC), cellulose (C) and lignin (L) with mono extract of PF within 10 days. The influence of mono extract of each strain (PF PS and PE) and their mixed extracts (PF+PS, PF+PE, PS+PE and PF+PS+PE) on degradation of cell wall constituents were remarkably differed. The mixed extract treatment proved maximum 13.6% HC loss by PF+PS+PE extract, 9.2% loss in C by PF+PS extract and 5.2% loss of L by the PF+PS+PE extract treatment. The highest dry matter loss (8.2%) was recorded with PF+PS+PE mixed extract combination.     

敌敌畏对鲫鱼的急性毒性及类球红细菌的解毒作用(Acute Toxicity of Dichlorvos to Carassius auratus and the Detoxification by Rhodobacter sphaeroides)

敌敌畏是使用最为广泛的有机磷农药之一,具有较高的水溶性,对水生动物的危害较大.为初步探讨水体中敌敌畏降解菌对中毒鱼类的解毒作用,测定了敌敌畏对鲫鱼(Carassius auratus)幼鱼的急性毒性,调查了一株敌敌畏高效降解菌——类球红细菌(Rhodobacter sphaeroides)的解毒作用.结果表明:敌敌畏对鲫鱼幼鱼48、72、96h的LC50分别为45.1、30.4、23.3mg·L-1,安全浓度为2.3mg·L-1,属低毒农药.在不同敌敌畏暴露浓度的养鱼水中添加类球红细菌(5×107CFU·mL-1)可以显著降低鲫鱼的死亡率,20mg·L-1和35mg·L-1敌敌畏暴露浓度下,鲫鱼96h死亡率分别由40%和100%降低至0和15%;利用液相色谱检测了敌敌畏在添加和不添加类球红细菌的养鱼水中的降解规律,发现类球红细菌的解毒作用主要是对水中敌敌畏的快速降解,降解过程中没有积累有毒代谢产物.     

黄杜鹃叶提取物对桔二叉蚜的毒力及其毒理研究

黄杜鹃(Rhododendron molle)叶提取物对桔二叉蚜(Toxoptera aurantii)有较强的毒杀作用．黄杜鹃叶的乙醇总提物对桔二叉蚜无翅成蚜的半致死浓度(LC50)为1558mg/L在黄杜鹃叶提取物的4种萃取物中，乙酸乙酯萃取物对桔二叉蚜的毒力最强，其LC50为364mg／L，表明黄杜鹃叶含有的杀虫活性物质主要存在于乙酸乙酯萃取物中．实验表明，黄杜鹃叶乙酸乙酯萃取物对桔二叉蚜的乙酰胆碱酯酶(AchE)和羧酸酯酶(CarE)有一定的抑制作用．桔二叉蚜经10000mg/L的黄杜鹃叶乙酸乙酯萃取物溶液处理24h后，其体内的乙酰胆碱酯酶的活性为对照组的0．6543倍，羧酸酯酶的活性仅为对照组的0．4021倍．图2表4参10     

Polyphenol contents of five of medicinal plants from Cameroon and effects of their extracts on antioxidant capacities of human breast cancer cells

The aim of the study was to investigate the antiproliferative properties of extracts of five plants on breast cancer cells (MDA-MB-231) and their effects on antioxidant activities. The total phenol contents of the extracts were determined to establish a correlation to their antiproliferative effects which were evaluated by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide viability assay. Enzymes involved in oxidative stress were also determined. Total polyphenol contents of the five extracts varied from 170 to 440 mg L^?1 of gallic acid per mg of extract with the highest value for the extract of Sida cordifolia. Cell growth inhibition was observed within 24 h, with inhibitory concentration values ranging from 7.3 to 25 mg L^?1 depending on plant extract. At concentrations between 100 and 200 mg L^?1, all the extracts exhibited significant reduction of cell proliferation in time-dependent and linear manner. The activities of superoxide dismutase and catalase of cells treated for 24 h with the extract of S. cordifolia were 3.5 U per mg protein and 8 mole H₂O₂ consumed per min per mg protein higher than those of cells treated with an extract of Viscum album.     

苯并［a］芘对黑鲷肝脏GST活性的影响及其与肝脏代谢酶和胆汁代谢产物之间的变化关系

为探讨苯并[a]芘(B[a]P)暴露对鱼类的影响并筛选特异、敏感的生物标志物,研究了B[a]P对黑鲷(Sparus macrocephalus)肝脏谷胱甘肽-S-转移酶(GST)活性的影响,并分析了肝脏7-乙氧基异吩唑酮-脱乙基酶(EROD)、GST活性与胆汁中B[a]P典型代谢产物3-羟基-苯并[a]芘(3-OHB[a]P)3者之间及与B[a]P之间的剂量、时间-效应关系.结果显示:在B[a]P(0.5、1.0、2.0和5.0μg·L-1)暴露期间,肝脏GST活性随暴露时间总体呈"钟"形的变化趋势,在第2d时达到峰值;在第12h、1d和2d时,GST活性与B[a]P暴露浓度呈显著正相关(R12h=0.966(p<0.05)、R1d=0.953(p<0.05)、R2d=0.824(p<0.05)).与前期研究结果对比分析发现,B[a]P(0.5、1.0、2.0和5.0μg·L-1)暴露7d时,胆汁中3-OHB[a]P浓度与B[a]P暴露浓度呈显著的剂量-效应关系(R=0.943,p<0.05);黑鲷肝脏GST、EROD活性与3-OHB[a]P的对数浓度均呈显著正相关(RGST=0.740(p<0.05)、REROD=0.839(p<0.05));2.0μg·L-1B[a]P暴露后,黑鲷肝脏EROD、GST活性与3-OHB[a]P随暴露时间的变化趋势基本相同,但并不完全一致.鱼类肝脏EROD、GST与胆汁中B[a]P代谢产物3者之间的变化关系较为复杂,暴露浓度和时间是影响其变化的重要因素,肝脏GST和EROD活性比胆汁中代谢产物更易受其影响,在B[a]P的环境监测中代谢产物可能是一种更为可行的生物标志物.     

信号分子对类球红细菌(Rhodobacter sphaeroides)碳源利用的影响

类球红细菌拥有cer群体感应系统.它既有细胞间信息传递的功能,也对碳源利用进行调节.为调查自诱导物对碳源利用的影响,在仅少量EDTA作唯一能源物的培养基中加入N-(tetradecanoyl)-DL-homoserine lactone,通过测定活菌数变化观察这种类球红细菌自诱导物的结构类似物对碳源利用的影响.然后利用Biolog-GN鉴定板,在加入外源信号分子和对照的条件下,研究了类球红细菌正常培养与饥饿培养下对95种单一碳源的不同利用情况.结果表明,在存在信号分子的饥饿情况下,细菌能更好的利用EDTA维持活菌数量.当培养到6h的时候,加入信号分子培养的鉴定板中,正常培养情况下细菌对碳源的利用丰富度指数最高;而在饥饿培养情况下细菌能利用更多种类的碳源.     

Toxicological and ecological implications of biotransformation enzymes in the tropical teleost Chaetodon capistratus

Hepatic cytochromes P450 (phase I monooxygenases) and glutathione transferases (phase II conjugating enzymes) were investigated in Chaetodon capistratus (Linnaeus) collected in Florida and Belize in June and December 1991, respectively. These biotransformation enzymes play major roles in the detoxification of xenobiotics by converting lipophilic chemicals to more hydrophilic, readily excretable metabolites. Content of total microsomal P450 (0.501 to 0.821 nmol mg^-1 microsomal protein) and rates of NADPH-cytochrome c (P450) reductase (270.7 to 330.2 nmol min^-1 mg^-1 microsomal protein) and glutathione transferase (2.81 to 3.12 g min^-1 mg^-1 cytosolic protein) in these fish were greater than in most untreated fish species, i.e., fish that have not been exposed to PAHs (polycyclic aromatic hydrocarbons) or PCBs (polycyclic biphenyls). Ethoxyresorufin O-deethylase (EROD) rates (0.029 to 0.171 nmol min^-1 mg^-1) were also comparable to those in most untreated marine fish. Immunoblot analysis with monoclonal antibody (MAb) 1-12-3 to scup P450E (the EROD catalyst and a teleost representative of the PAH-inducible CYP1A gene subfamily) showed slight amounts of cross-reacting protein in C. capistratus liver microsomes. Hepatic CYP1A content and EROD activity did not differ significantly between fish collected in Florida and Belize, suggesting that the two sites differed little in contamination by CYP1A inducers. Immunochemical analyses with polyclonal antibodies to scup P450B (a teleost representative of the CYP2B subfamily) and human CYP3A4 cross-reacted strongly with C. capistratus hepatic proteins. The CYP2B and CYP3A subfamilies in mammals are believed to have partially evolved in response to toxic dietary allelochemicals. C. capistratus regularly feeds on terpenoid-rich gorgonian corals, suggesting that biotransformation enzymes may be involved in the metabolism of dietary allelochemicals as well as anthropogenic xenobiotics in this species.     

Effect of trichloroethylene (TCE) inhalation on biotransformation enzymes of rat lung and liver

Trichloroethylene (TCE) is widely used as an industrial solvent and cleaning fluid. In the present study the toxic effects of TCE inhalation on pulmonary and hepatic biotransformation enzymes in rats have been investigated by assay of aniline hydroxylase (AH), aminopyrine-N-demethylase (APD), benzo-a-pyrene hydroxylase (BH) and glutathione-s-transferase (GST) activities and glutathione (GSH) contents in liver as well as lungs of exposed animals. In both organs phase I and phase II drug metabolizing enzymes have been found to be increased along with decrease in GSH contents following TCE inhalation. Pulmonary as well as hepatic MFO's seem to be activated by inhaled TCE probably in an attempt for its rapid detoxification and reduced glutathione is used during its biotransformation.     

Effects of Population Size, Mating System, and Evolutionary Origin on Genetic Diversity in Spiranthes sinensis and S. hongkongensis

Hong Kong once supported more than 109 species of wild orchids, of which approximately 30% were endemic. Most of the local wild orchids have now become rare or endangered. I conducted a comparative study of genetic diversity in two closely related terrestrial orchids, an allotetraploid, Spiranthes hongkongensis , and its diploid progenitor, S. sinensis , to assess the effects of the population bottleneck associated with the origin of the polyploid and to investigate the relationships between number of breeding individuals, mating system, and level of isozyme variation in their populations. Nearly complete genetic uniformity was observed both within and among populations of S. hongkongensis . In contrast, S. sinensis had high levels of genetic variation for all of the genetic parameters examined. Regression analysis of population size and several components of genetic diversity in S. sinensis revealed that, among various measures of within-population variation, the proportion of polymorphic loci ( P ) and average number of alleles per locus ( A ) or per polymorphic locus ( A _p ) were the most sensitive to population size ( R ² = 0.942, p = 0.001; R ² = 0.932, p = 0.002; and R ² = 0.923, p = 0.002 respectively). The highly negative correlation ( r = −0.999, p < 0.01) between population size and the mean frequency of private alleles in pairwise population comparisons, p (1), indicated that population size may also be used to predict the extent of population differentiation caused by random genetic drift. Conservation of genetic diversity in S. sinensis could be maximized by protecting several of both large and small populations, whereas fewer populations may be needed to achieve this goal for S. hongkongensis.     

Biotransformation rates of xenobiotic compounds in relation to enzymes activities: A critical review

Many biotransformation reactions and many activities of enzymes involved in the biotransformation of xenobiotics in fish have been described. Therefore, the prediction of biotransformation of (other) xenobiotics through literature data seems to be possible. However, very few data allow correlation of in vitro enzyme activities with in vivo biotransformation rates. This is mainly due to a lack of quantitative data. Suggestions are made for future research, which may help to relate enzyme activities and biotransformation in fish.