重金属对蚯蚓的毒性毒理研究

通过室内模拟实验以及对株洲市重金属污染区农田土壤中采样，进行了重金属对蚯蚓的毒性毒理研究．毒性实验表明，各重金属元素４８ｈ的ＬＤ５０分别为：Ｃｄ１０００ｍｇ／ｋｇ，Ｐｂ８１２ｍｇ／ｋｇ，Ｃｕ６３３ｍｇ／ｋｇ，Ｈｇ３０４ｍｇ／ｋｇ，Ｚｎ５２８ｍｇ／ｋｇ，Ｃｒ４２８ｍｇ／ｋｇ．对蚯蚓毒理研究证明，重金属对蚯蚓酯酶同工酶有抑活作用，而重金属对蚯蚓过氧化物酶同工酶有激活作用．在光镜下观察，可见重金属污染可引起蚯蚓的体表溃疡及产生肿瘤，可使胃肠道粘膜层出血、背血管肿胀，同时可引起胃肠道粘膜上皮细胞产生萎缩或溃疡灶．蚯蚓特有的黄色细胞团污染程度不同而显示出消长规律．在电镜下可见到在重污染区的蚯蚓胃肠道粘膜上皮细胞中的ｒＥＲ肿胀、高尔基体膨大，线粒体嵴消失，甚至空泡化或解体．另可见细胞核膜间隙肿胀、断裂、核质外溢，胞质自溶．蚯蚓以上病变的大小是随土壤重金属污染区的污染级别而定；各病理变化指标均能定性地反映土壤重金属污染状况．     

表面活性剂与Pb、Zn协同作用对蚕豆叶细胞的损伤

采用透射电子显微镜及能量发散X-射线微量分析技术,研究了非离子型表面活性剂烷基酚聚氧乙烯醚(AE)与矿山尾砂中高浓度Pb、Zn协同作用对蚕豆叶细胞的损伤.结果表明,高浓度Pb、Zn使蚕豆叶绿体中类囊体解体,嗜锇颗粒的数量和体积显著增加.细胞外间隙和液泡中出现大量的不溶性毛发状晶体.AE与Pb、Zn复合污染时,细胞内膜结构的透性增加,细胞内毛发状晶体扩散到叶绿体、细胞核中.因此,AE可加重Pb、Zn对蚕豆叶细胞的损伤作用,毛发状晶体的形成可能与细胞脱毒有关.     

皱纹盘鲍"足萎缩症"幼鲍超微结构研究

利用超薄切片电镜技术对2005年春季大连养殖场患"足萎缩症"皱纹盘鲍(Haliotis discus hanni)幼鲍足、吻、外套膜、肝胰腺超微结构进行了研究,结果表明:足肌肉组织、吻组织、外套膜组织细胞内出现典型的病理变化.肠组织细胞内出现外膜包裹的聚集大量α糖原颗粒的包涵体.鳃组织细胞内线粒体形态异常,含有类晶格微丝结构,病变细胞间出现α糖原颗粒累积包涵体,说明病变细胞内出现严重的代谢障碍.肝胰腺组织损伤最为严重,超微结构显示为大量空泡结构和淀粉样物质,已无可识别的细胞结构.同时在足肌肉组织细胞间隙发现大量的直径为45～55 nm的病毒样颗粒,在吻细胞肿胀的内质网中发现80～110 nm的病毒样颗粒,具有囊膜,囊膜厚度约为2 nm.     

Effects of chitosan on growth of an aquatic plant (Hydrilla verticillata) in polluted waters with different chemical oxygen demands

Effects of chitosan on a submersed plant, Hydrilla verticillata, were investigated. Results indicated that H. venicillata could prevent ultrastructure phytotoxicities and oxidativereaction from polluted water with high chemical oxygen demand (COD). Superoxide dismutase (SOD) activity and malondialdehyde (MDA) contents in H. verticillata treated with 0.1% chitosan in wastewater increased with high COD (980 mg/L) and decreased with low COD (63 mg/L), respectively. Ultrastructural analysis showed that the stroma and grana of chloroplast basically remained normal. However, plant cells from the control experiment (untreated with chitosan) were vacuolated and the cell interval increased. The relict of protoplast moved to the center, with cells tending to disjoint. Our findings indicate that wastewater with high COD concentration can cause a substantial damage to submersed plant, nevertheless, chitosan probably could alleviate the membrane lipid peroxidization and ultrastructure phytotoxicities, and protect plant cells from stress of high COD concentration polluted water.     

杀虫单对不同倍性泥鳅鳍细胞系的毒性作用

以泥鳅鳍二倍体(DIMF)和三倍体(TRMF)细胞系为受试细胞,研究杀虫单对2种细胞系的毒性作用。采用噻唑蓝(MTT)法测得DIMF与TRMF 24 h半致死浓度分别为119.73 mg·L-1、146.26 mg·L~(-1)。DIMF的敏感性明显高于TRMF。经杀虫单处理的活体细胞表现为细胞伸长,空泡化和脱落并游离于培养基表面的现象。2种细胞系酶活力测定的结果显示:杀虫单浓度为0~100 mg·L~(-1)时,SOD和GST活力随着浓度的增加而增加,100~200 mg·L~(-1)浓度组酶活力逐渐降低;0~200 mg·L~(-1)时,ACh E活力与杀虫单浓度呈负相关,并且三倍体3种酶活力均高于二倍体。微核试验结果显示:50 mg·L~(-1)杀虫单就能对细胞核造成损伤并形成微核,微核率随杀虫单浓度增加而增加。当杀虫单浓度达到200 mg·L-1时,微核率出现最大值,DIMF和TRMF分别为3.3‰和3.7‰,2种细胞的测试结果无显著性差异(P0.05)。电镜观察结果显示:对照组DIMF和TRMF超微结构无明显差异;DIMF和TRMF病理变化情况相似:染色质凝集趋边,细胞核分解成多个,细胞内出现空泡和凋亡小体,表现出凋亡的特征。研究表明杀虫单的细胞毒性机制是通过改变细胞内酶活性从而诱导凋亡,不同倍性细胞系之间的差异主要与多倍体细胞体积大,胞内物质多,分裂快,生长旺盛等有关。     

铜绿微囊藻溶藻菌EA-1的分离鉴定及溶藻特性

从广州流花湖分离获得一株溶藻菌株EA-1,16S rDNA分析表明菌株EA-1属于肠杆菌属（Enterobacter）.研究了肠杆菌EA-1对铜绿微囊藻的溶藻效果和溶藻机制.结果表明,对数期EA-1具有最佳溶藻效果,投加比例为10%,初始叶绿素a含量为1.43mg/L时,EA-1能实现3d内完全除藻,叶绿素a含量为2.39mg/L时,共培养6d后,抑制率为84.1%±1.3%.EA-1通过分泌胞外溶藻物质间接溶藻,生理生化响应表明,EA-1无菌滤液胁迫下,藻细胞膜脂过氧化损伤严重,超氧化物歧化酶（SOD）、过氧化氢酶（CAT）和抗坏血酸过氧化物酶（APX）活性先急剧上升后下降.三维荧光光谱（EEM）表明溶藻产物为类腐殖酸,类富里酸和类蛋白类物质.扫描电子显微镜（SEM）显示藻细胞出现褶皱,内陷和萎缩现象.透射电子显微镜（TEM）显示藻细胞破坏过程为：首先,胶质层与细胞壁分离,光合片层变得松散和不规则,内含物被部分降解.随后,光合片层被彻底破坏,DNA核物质和多聚磷酸盐等营养物质颗粒被降解,藻细胞内部结构被完全破坏,藻细胞死亡.     

Iron (Fe2+)-induced toxicity produces morphological and physiological changes in roots in Panax ginseng grown in hydroponics

ABSTRACT

Rusty roots markedly influence on ginseng cultivation, and this phenomenon often attributed to iron (Fe) induced toxicity. To examine the physiological mechanisms underlying Fe-initiated toxicity as evidenced by rusty roots in Panax ginseng, morphological and physiological changes in roots were investigated in hydroponics using Fe²⁺ concentrations of 50 (control), 100, 200, 400 or 600 µM. Compared with control, reddish-brown deposition at the root surface increasingly appeared as Fe²⁺ concentration increased (≥200 µM). The pH also rose as Fe levels were elevated. Higher external Fe²⁺ concentrations produced changes in root organelles and cell structures. Structural alterations in mitochondria due to excess Fe storage, protoplast shrinkage and cell vacuolation as well as formation of central vacuole with deposits in roots were observed. In addition, apparent cell wall thickening, cell wall folding and shrinkage, damage of cell membranes and a large amount of cell debris occurred at higher external Fe²⁺ concentrations (≥400 µM). The Fe²⁺ mediated damage resulting in morphological and physiological changes in ginseng roots was concentration and pH dependent.     

Ultrastructural changes in the rabbit liver induced by carbamate insecticide bendiocarb

Carbamates (CB) are used as insecticides and some of them have been registered as human drugs. The mechanism of CB poisoning involves reversible inhibition of acetylcholine esterase. In the present study, we investigated changes in liver ultrastructure in rabbits (Oryctolagus cuniculus) which were administered bendiocarb for 3, 10, 20, and 30 days. Rabbits in all experimental groups received capsules of bendiocarb (96% Bendiocarb, Bayer, Germany) per os daily at a dose of 5 mg/kg of body weight, and after day 11 received the same dose every 48 h. The observed changes were only moderate, focal, and the effect on the liver was not uniform. On the third day of the experiment, injured hepatocytes had dilated bile capillaries with reduced microvilli. There were no visible alterations in the intercellular contacts. Nuclei of these cells were irregular in shape. Many hepatocytes showed considerable increase in the number of peroxisomes. On day 10 of the experiment, the number of peroxisomes was reduced. Other changes, such as dilated rough endoplasmic reticulum and proliferation of smooth endoplasmic reticulum were observed on day 20. The number of lipid droplets in hepatocytes gradually increased. Usually they were present in low numbers, but on day 30 of the experiment their number increased significantly. They coalesced and formed a single lipid droplet which changed the shape of the nuclei. The results presented in this study indicate that both short and long-term administration of bendiocarb affects the liver ultrastructure. At the same time we also observed rapid onset of regeneration of the damaged tissue through activation of hepatocytes and oval cells.     

苯并[a]芘对栉孔扇贝(Chlamys farreri)组织显微和超微结构的影响

在实验条件下研究了苯并[a]芘(B[a]P)暴露对栉孔扇贝的鳃、消化盲囊显微(10d、20d)和超微结构(10d)的影响.观察结果表明,10μg·L-1B[a]P处理10d时,栉孔扇贝的鳃上皮粘液细胞增多,在回血管中血细胞出现聚集现象;消化盲囊腺泡中嗜碱性颗粒增多,腺泡内细胞间界限模糊.超微结构观察可见,鳃上皮纤毛和微绒毛脱落,鳃上皮细胞内次级溶酶体增多,核周池扩大,鳃血腔中出现血细胞坏死后残留的细胞核;在消化盲囊分泌细胞中线粒体嵴减少,出现线粒体缺失形成的空泡,细胞内质网片断化,核糖体脱落.由10μg·L-1B[a]P处理20d时,在鳃回血管中发现血细胞坏死后残留的固缩细胞核,鳃丝的上皮细胞排列不规则甚至断裂,部分消化腺泡崩解,鳃丝结构和消化导管上皮结构损害严重;消化盲囊组织结构的损伤比鳃更为严重.这些组织结构损伤表明,栉孔扇贝在B[a]P胁迫下组织学的变化特征与过程与贝类组织解毒代谢过程相符合.     

碲化镉量子点对小鼠肝脏超微结构及重要生物酶活性的影响

探讨了碲化镉量子点(cadmium telluride quantum dots,CdTeQDs)对小鼠肝组织超微结构及琥珀酸脱氢酶(succinate dehydrogenase,SDH)和三磷酸腺苷酶(adenosine triphosphatase,ATPase)活性的影响。将20只雄性ICR小鼠随机分成QDs染毒组和对照组,采用尾静脉注射方式进行一次性染毒,染毒组每只小鼠注射2μmol·kg-1体重的CdTeQDs(以Cd~(2+)的摩尔浓度计算)。每5只为一组分别在染毒后的1 d、3 d和7 d处死小鼠,另外5只注射生理盐水作为对照组。取部分肝组织在透射电镜下观察其超微结构变化并使用试剂盒测定肝脏中SDH、Na~+/K~+-ATP酶、Ca~(~(2+))/Mg~(2+)-ATP酶的活性。结果显示CdTeQDs暴露后染毒组小鼠肝细胞胞浆疏松,线粒体出现肿胀、空泡、嵴减少等结构变化,表明肝细胞受到损伤且染毒1 d后损伤最严重;肝脏中染毒组SDH活性与对照组相比显著降低(P0.05);染毒1 d后,Na~+/K~+-ATP酶和Ca~(2+)/Mg~(2+)-ATP酶活性与对照组相比显著升高,随着染毒后时间的延长,其活性呈现下降的变化趋势。单次量注射CdTeQDs能够引起小鼠肝脏损伤,抑制SDH活性,引起ATP酶活性改变,这些症状在暴露时间内呈现恢复趋势。