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1.
The in vitro acute cytotoxicity of the neonicotinoid insecticide acetamiprid to FG cell line, derived from the gill of flounder Paralichthys olizaceus, was tested by the three widely used endpoint bioassays-neutral red （NR） assay, tetrazolium （MTT） assay and cell protein assay. It was found that acetamiprid was increasingly toxic to FG cells at concentrations of 1 μg/cm^3 or above, and the inhibitory concentration 50% values for NR, MTF, and cell protein assays were 38.38, 36.27 and 32.03 μg/cm^3, respectively. This appeared to be the first report on the in vitro cytotoxicity of acetamiprid to non-mammalian vertebrate cells. Ultrastructural examination revealed that for the cells exposed to 60 μg/cm^3 acetamiprid for 48 h, their mitochondria were severely damaged with the cristae swelled up or disrupted, while their nuclei and rough endoplasmic reticlum （RER） appeared to be still normal. This suggests that mitochondria are possibly the primary target of acetamiprid.  相似文献   

2.
1 Introduction Forthe lastm ore than 20 a,ow ing to the develop-m ent of observation and experim ent technologies form arine m icroorganism s,m arine biologistshave discov-ered m any m icroorganism s w hich are difficult to beobserved by com m on m icroscope before. T hus, thecognition to the m arine phytoplankton com m unitiesand the structure of food chains has achieved im por-tantprogress.E specially,w ith the developm entand ap-plication of epifluorescence m icroscopy and flow cy-tom etry…  相似文献   

3.
Benzothiazole, a common chemical associated with tire manufacturing and industrial wastewater, is a principal component of both fresh water and estuarine tire leachate, a neurotoxicant to larval sheepshead minnows (Cyprinodon variegatus) in in vivo estuarine studies. The neurotoxic potential of benzothiazole was investigated following in vivo and in vitro exposure of sheepshead minnows to 3.75, 7.5, 15, 30 and 60 mg/l benzothiazole. Following benzothiazole exposure, fish were evaluated for survival, growth and histological alterations. Fish mortality occurred after 5 days of exposure to 60 mg/l (LC50 = 41.9). Significant decreases in larval growth were noted at all concentrations. Histologically, gills had cellular alterations but the central nervous system lacked the severe cellular damage seen in previous tire leachate exposure studies. Benzothiazole cytotoxicity to primary cultures of brain cells from sheepshead minnow and tilapia (Oreochromis niloticus) and two epithelial cell lines was evaluated using a tetrazolium salt assay (MTT) at 1 and 4 days. In vitro results indicate primary cultures of brain cells are less sensitive to benzothiazole than epithelial cell lines. Significant cytotoxicity to the epithelial cell lines was noted at 30 and 60 mg/l concentrations. Histologically and cytotoxicologically, the present study indicates that benzothiazole is a gill toxicant and not a neurotoxicant.  相似文献   

4.
雪卡毒素(ciguatoxin)是一类具有神经毒性的海洋藻类毒素,高纯度雪卡毒素是开展其相关研究的物质基础。以石斑鱼肝脏为原料,对其脂溶性粗提物经Florisil柱和Sephadex LH-20柱纯化后,用高效液相色谱/质谱(HPLC/MS)和细胞毒性方法对其进行鉴定,证明为太平洋雪卡毒素(P-CTX-1)。HPLC鉴定分析提取物与太平洋雪卡毒素(P-CTX-1)具有相同的保留时间。HPLC/MS检测表明纯化样品在m/z=1 111.3和m/z=1 133.0处出现雪卡毒素[M+H]+峰和[M+Na]+峰,与已有的P-CTX-1分子离子峰完全吻合。四甲基偶氮唑蓝法(MTT)细胞毒性试验证明了雪卡毒素样品对人神经母细胞瘤(SK-N-SH)具有毒性作用,毒素剂量与细胞生长抑制率成线性关系。细胞毒性试验进一步确认该提取物为雪卡毒素,且纯度较高。  相似文献   

5.
利用免疫细胞化学法测定鱼类淋巴囊肿病毒(LCDV)滴度.以牙鲆鳃细胞系(FG)作为感染细胞,将生长旺盛的FG细胞接种于48孔培养板中培养至形成细胞单层,用2倍连续稀释的LCDV粗提液分别接种FG细胞.固定各稀释度LCDV感染后的FG细胞,孵育抗牙鲆LCDV单克隆抗体,其后再运用生物素-亲合素反应系统,以碱性磷酸酶底物APRed试剂盒发色.倒置显微镜观察,被病毒感染的FG细胞的细胞质呈现红色,未被感染细胞的细胞质呈无色.记录各稀释度病毒感染的阳性细胞孔数,按Reed-Muench法计算组织细胞培养半数感染量(TCID0).结果显示,免疫细胞化学法测得LCDV在FG的滴度为1.77×210 TCID50/mL.该法可以用来有效测定LCDV滴度,且结果直观、准确性较好,灵敏度较高.  相似文献   

6.
节旋藻和螺旋藻对7种抗生素敏感性的比较研究   总被引：7，自引：1，他引：6  
对两种丝状蓝藻(钝顶节旋藻和盐泽螺旋藻)在基因工程中常用作选择试剂的7种抗生素——氯霉素、氨苄青霉素、红霉素、链霉素、卡那霉素、庆大霉素和新霉素的敏感性作比较实验.结果表明,两种蓝藻对抗生素的敏感性既有共同的特点,也有明显的差异.它们对红霉素、氯霉素和链霉素最敏感,致死浓度分别为0.1,0.5和5μg/cm3.两种蓝藻对氨苄青霉素比较敏感,1μg/cm3的氨苄青霉素即可抑制Arthrospira.341和Spirulina.351的生长,但6d后生长恢复.Arthrospira.341和Spirulina.351对卡那霉素、庆大霉素和新霉素均有抗性,而且存在很大差异:300μg/cm3的卡那霉素对Arthrospira.341的生长仍然没有影响,但对于Spirulina.351,50μg/cm3的卡那霉素即对其生长有明显抑制作用;200μg/cm3的卡那霉素即可将其全部致死.200μg/cm3的庆大霉素和300μg/cm3的新霉素不能抑制Arthrospira.341和Spirulina.351的生长,但在这两种抗生素环境中两种藻的生长状态有很大差异.并验证了氯霉素、红霉素和链霉素是节旋藻和螺旋藻基因转化过程中的有效的抗性选择剂,也从对抗生素敏感性方面表明节旋藻和螺旋藻两个属的遗传差异.  相似文献   

7.
Chemical investigation of the ethanol extract of the marine green alga Chaetomorpha basiretorsa Setchell led to the isolation of a new sterol stigmast-4,28-dien-3α,6β-diol 1 in addition to the five known sterols of β-lawsaritol 2, saringosterol 3, 24-hydroperoxy-24-vinyl - cholesterol 4, β-stigmasterol 5, 29-hydroxystigmasta-5, 24 （28） -dien-3β-ol 6. Compounds were isolated by normal phase silica gel and Sephadex LH - 20 gel colum chromatography, reverse phase HPLC and recrystalization. Their structures were elucidated by spectroscopic methods including MS, IR 1D/2D NMR and X-ray analysis. Cytotoxicity of compounds was screened by using the standard MTF method. All these compounds were isolated from the green alga Chaetomorpha basiretorsa Setchell for the first time and they were inactive （50% inhibitory concentration was greater than 10 μg/cm^3） against KB, Bel -7402, PC - 3M, Ketr 3 and MCF-7 cell lines.  相似文献   

8.
Antitumor activity is one characteristic function of some certain antimicrobial peptides(AMPs) found in recent years. In the present study, we attempted to detect potential anticancer activity of a recombinant piscidin 5-like from Larimichthys crocea(r Lc-P5L) which owned widely antibacterial and strong antiparasitic activity in vitro.The light microscope observation indicated r Lc-P5L was of antitumor activity to He La cells, 293 T cells and L929 cells. MTT assay showed the toxic sensitivity of...  相似文献   

9.
Lack of shrimp cell lines has hindered the study of pollutants which adversely affects shrimp health and its export value. In this context a primary haemocyte culture developed from Penaeus monodon was employed for assessing the cytotoxicity and genotoxicity of two heavy metal compounds, cadmium chloride and mercuric chloride and two organophosphate insecticides, malathion and monocrotophos. Using MTT assay 12?h IC(50) values calculated were 31.09?±?16.27?μM and 5.52?±?1.16?μM for cadmium chloride and mercuric chloride and 59.94?±?52.30?mg?l(-1) and 186.76?±?77.00?mg?l(-1) for malathion and monocrotophos respectively. Employing Comet assay, DNA damage inflicted by these pollutants on haemocytes were evaluated and the pollutants induced DNA damage in >60% of the cells. The study suggested that haemocyte culture could be used as a tool for quantifying cytotoxicity and genotoxicity of aquaculture drugs, management chemicals and pollutants.  相似文献   

10.
于红  吕锐  张学成 《海洋科学》2008,32(1):38-40
采用MTT法(四甲基偶氮唑盐比色法)研究了钝顶螺旋藻多糖(PSP)对Hela细胞及HepG2细胞生长的抑制作用。结果表明,随PSP浓度及培养时间的增加,肿瘤细胞存活率逐渐降低,抑制率逐渐增加,以PSP40 mg/L作用72 h最为显著。应用Annexin V/PI双染色流式细胞仪检测了早期Hela细胞凋亡,未经PSP处理的正常Hela细胞凋亡细胞极少,经PSP处理的细胞,凋亡细胞的百分比明显高于正常对照组,其作用随着剂量的增加和时间的延长而增强,具有量效关系和时效关系。结果说明PSP的抗肿瘤机制,除了诱导肿瘤细胞凋亡,还存在细胞毒性等其他机制,是多种机制共同作用的结果。  相似文献   

11.
The effects of ibuprofen (IBU) on the immune parameters of the clam Ruditapes philippinarum were investigated after a 7-day exposure to sublethal IBU concentrations (0, 0 + ethanol, 100, 500, and 1000 μg/L). Total haemocyte count (THC), haemocyte diameter and volume, haemocyte proliferation and uptake of the vital dye Neutral Red (NR) were measured. The cytotoxicity (assessed by the lactate dehydrogenase assay, LDH) and the capability of IBU to induce DNA fragmentation (indicative of apoptosis) were also investigated. The exposure of clams to the highest IBU concentration significantly reduced their THC, whereas no significant changes were observed in either the diameter or volume of haemocytes. Significant increases in haemocyte proliferation were recorded in clams that were exposed to the two highest tested concentrations of IBU. Exposure of clams to 1000 μg IBU/L significantly reduced NR uptake and increased haemolymph LDH activity. Conversely, IBU did not induce DNA fragmentation in haemocytes. Although the IBU concentrations tested in this study were higher than those generally recorded in aquatic environments, results obtained indicate that exposure of clams to IBU induces significant alterations in the immune parameters and suggest potential immunosuppression in treated clams.  相似文献   

12.
研究革皮氏海参(Pearsonothuria graeffei)皂苷单体化合物echinoside A(EA)和ds-echinoside A(DSEA)对血管新生的抑制作用.采用MTT法和AO/EB荧光染色法研究EA和DSEA对人脐静脉血管内皮细胞Human Umbilical Vein Endothelial Cells(HUVEC)增殖、凋亡的影响;采用小管形成实验观察EA和DSEA对HUVEC分化形成小管能力的影响;细胞间粘附实验比较研究EA和DSEA对HUVEC细胞同肿瘤细胞之间粘附的影响;鸡胚绒毛尿囊膜(CAM)血管新生模型,观察EA和DSEA抑制CAM血管新生的情况.实验结果显示,EA和DSEA能够显著抑制HUVEC细胞的增殖(P＜0.05,P＜0.01),并诱导其凋亡;在体外能够显著抑制小管的形成能力,可显著抑制同肿瘤细胞间的粘附作用,在体内能够减少CAM新生血管的分支数目,而且DSEA的活性要强于EA.提示草皮式海参皂苷单体EA和DSEA都能够显著抑制肿瘤血管的新生,其活性与结构相关.  相似文献   

13.
Boleophthalmus pectinirostris is an amphibious economic fish and wildly distributed in the southeast coast of China. In this study, Aroclor 1254 was intraperitoneally injected into B. pectinirostris with 1, 2 and 4 μg/(g·d) for 28 d to assay the reproductive organ weight, the sperm quality (sperm concentration and motility), and the testicular mitochondrial testicular mitochondria oxidative stress. The results show that the sperm number and motility in seminal vesicles, the absolute weight of testes and seminal vesicles of B. pectinirostris treated with 2 and 4 μg/(g·d) Aroclor 1254 decreased significantly as compared to the controls (p <0.05), while those treated with 1 μg/(g·d) Aroclor 1254 had no significant effects on these indictors. For the relative weight of reproductive organs, significant reduction (p <0.05) was only observed in the seminal vesicles of B. pectinirostris treated with 4 μg/(g·d). SOD activities and GSH levels in all the Aroclor 1254 treatments were significantly lower than those of the controls (p <0.05). The activities of CAT, GPx, GR and the levels of Vit C also decreased significantly in comparison with the controls (p <0.05) at the higher dose of 2 and 4 μg/(g·d) Aroclor 1254 treatments. In addition, both H2O2 level and MDA content in testicular mitochondria of B. pectinirostris had a close correlation with Aroclor 1254 dosage, and were significantly higher than the controls (p <0.05). Those indicate that Aroclor 1254 can induce the oxidative stress of testicular mitochondria, and impair the reproductive function of male B. pectinirostris.  相似文献   

14.
The analysis of 103 samples collected quantitatively from the southern Taiwan Strait (22°4'-24°3'N, 117°5'-119°9'E), China, in August 1997 indicates that the abundance of heterotrophic nanoflagellates (HNF) ranges from 391 to 1 846 ×103 cell/dm3, with an average of 949 × 103 cell/dm3, and 96.2% of cells are in size of 2-22 μm in all HNF met in the samples. The HNF cells are 9.83-45.79 μg/dm3 after conversion from cell number to carbon content, with an average of 22.82 μg/dm3 . The HNF abundance is relatively low in comparison with other areas. The nitrogen and phosphorus nutrients of the water are lower than previous investigations, which may be caused by the El Nino. The biomass of HNF is higher in southern waters, where cell concentration is 3-5 folds higher than that in other waters, and nearly 2-fold as high as the average of the investigated areas. The HNF is mainly habitat in 0-30 m water layer. Both horizontal and vertical HNF distribution must be influenced by hy-drographical dynamic pro  相似文献   

15.
Vertical distributions of phytoplankton biomass, compositions and size structure were investigated during the spring-intermonsoon (April 22 to 30) of 2010 along transact 10°N of the Bay of Bengal, northern Indian Ocean. Surface phytoplankton biomass (Chl a) was (0.065§0.009) μg/L, being greater than 80% of which was contributed by pico-phytoplankton (<3 μm). The Chl a concen- tration vertically increased to the maximal values at deep chlorophyll maximum (DCM) layer that shoaled eastwards from 75 to 40 m. The Chl a biomass at DCM layer generally varied between 0.2 and 0.4 μg/L, reaching the maximum of 0.56 μg/L with micro-phytoplankton cells (>20 μm) accounting for 58% and nano- (3-20 μm) or pico-cells for 15% and 27%, respectively. In particular, the cells concentration coupling well with phosphate level was observed at middle layer (75-100 m) of 87° to 89°E, dominated by micro-cells diatoms (e.g., Chaetoceros atlanticus v. neapolitana, Chaetoceros femur and Pseudonitzschia sp.) and cyanobacteria (i.e., Trichodesmium hildebrandtii), with the cells concentration reached as high as 4.0×104 and 4.3×104 cells/L. At the rest of the trans- act however, dino°agellates (e.g., Amphisdinium carterae and Prorocentrum sp.) were the dominant species, with the cells concentration varying from 0.3×103 to 6.8×103 cells/L. Our results also in- dicate that the regulation of large cells (micro-, nano-) on phytoplankton biomass merely occurred at DCM layer of the Bay.  相似文献   

16.
本文旨在研究黄鳍金枪鱼(Thunnus albacares)碎肉中4种不同蛋白酶酶解物的3k Da超滤组分对RAW264.7细胞的免疫活性。以RAW264.7细胞为实验模型, 采用噻唑蓝比色法检测细胞增殖活性, 中性红吞噬实验检测细胞吞噬能力, Griess试剂检测细胞培养上清液中NO含量, 并分析酶解物氨基酸组成和分子量分布。实验结果表明, 胰蛋白酶酶解物在12.5~200μg·mL-1范围内无明显细胞毒性, 能增强RAW264.7细胞吞噬能力, 浓度为400μg·mL-1时达到最大值132.50%(P<0.01), 浓度在50~200μg·mL-1范围内能促进巨噬细胞NO释放(P<0.01), 且NO释放量与酶解液浓度存在一定的量效关系; 胰蛋白酶酶解物中必需氨基酸(essential amino acid, EAA)含量最高, 为51.09%, 其碱性氨基酸组氨酸、赖氨酸和精氨酸的含量分别为11.59%、11.00%和6.90%, 均高于其他蛋白酶酶解物, 并且分子量小于1k Da的小肽含量高达97.48%。研究结果表明, 4种蛋白酶酶解物中, 黄鳍金枪鱼碎肉胰蛋白酶酶解物的3k Da组分具有较明显的免疫活性, 这为其作为免疫活性肽的开发利用提供了理论依据。  相似文献   

17.
为建立来源于锦鲤的细胞系,本文采用组织块法,对来源于锦鲤(Cyprinus carpio)鳍条、吻端、肌肉、心脏、鳔、肠道、卵巢等组织的细胞进行原代培养和传代培养.在25℃下,添加20％胎牛血清、0.2μg/mL表皮生长因子(EGF)和25ng/mL成纤维生长因子(FGF)的L-15培养基中进行培养.结果显示,鳍条、心脏、鳔、吻端、肌肉、肠道、卵巢分别在原代培养第3、6、12、13、15、3、3天,有细胞从组织块迁出;对长至单层的鳍条、心脏、肌肉细胞进行传代培养,鳍条细胞已传至第39代,心脏和肌肉细胞分别传到第2代和第4代;鳍条、心脏和肌肉细胞分别呈现上皮细胞样、上皮细胞样和上皮细胞样与成纤维状混合型.第6代锦鲤鳍细胞的染色体计数,结果显示,细胞染色体数目分布范围为55～154条,2n=100.病毒敏感性实验,发现锦鲤鳍条细胞对草鱼出血病病毒(GCRV)和鲤春病毒血症病毒(SVCV)都敏感,且在24h内出现细胞病变(CPE);但对斑点叉尾鮰病毒(CCV)不敏感.锦鲤鳍条细胞系的建立为后期建立更多的锦鲤细胞系和鱼类病毒研究奠定了基础.  相似文献   

18.
圆紫菜人工色素突变体的诱导与分离   总被引：1，自引：0，他引：1  
为获得圆紫菜人工色素突变体，本文使用一定剂量的紫外线辐照其叶状体，培养数天后，在叶状体上出现了少量颜色变异细胞，它们呈斑点状无规则地分布在野生型细胞中间。再培养2~3周，这些色素变异细胞分裂形成了不同颜色的细胞块，其颜色呈浅绿黄、橄榄色、草绿、绿褐、黄褐、浅褐、深紫红和浅紫红色等。在辐照强度0~80 μW/cm2范围内，叶状体上色素变异细胞块出现的频率随辐照强度的增加而增加，但增加至80 μW/cm2以上时，随着辐照强度的增加，色素变异细胞块出现的频率反而下降，这表明80 μW/cm2为有效的辐照强度。将色素变异细胞块切下，置入充气瓶内充气培养，待其释放出单孢子，随后从单孢子萌发体中挑选出了黄褐、橄榄色、红色、褐红等纯色的突变体，并利用叶状体单性生殖分别获得它们的遗传纯合丝状体（品系）。各突变品系的F1叶状体与各自母体的颜色一样，说明其颜色是可稳定遗传的。与野生型品系相比，各色素突变品系的F1叶状体的生长速度、活体吸收光谱、3种主要光合色素和色素蛋白的含量及它们相互间的比值均发生了明显改变。  相似文献   

19.
于2007年4月至2008年3月对大亚湾大鹏澳海域表层沉积物中浮游植物休眠体进行了每月1次的周年调查.本调查分析鉴定出浮游植物休眠体79种,包括甲藻孢囊50种,硅藻休眠体26种.浮游植物休眠体丰度没有明显的季节变化规律,变化范围为217～6 764 cells/cm3,平均为1 376 cells/cm3;鱼类养殖区休眠体丰度较高,平均丰度为1 999 cells/cm3.浮游植物休眠体在数量上以硅藻为主,甲藻次之,而在鱼类养殖区也出现了较高的蓝藻休眠体.硅藻休眠体的平均丰度为783 cells/cm3,百分比含量为56.9%,从而反映了该海域硅藻型浮游植物群落结构.甲藻孢囊丰度的变化范围在79～1 237 cells/cm3,位于湾口对照区丰度较高,年平均为733 cells/cm3,而鱼类养殖区和贝类养殖区平均值分别为502、388 cells/cm3.异养型甲藻孢囊的种类和数量均比较丰富,其百分比含量为26.7%～82.4%.研究结果显示,在富营养化程度较高的养殖区海域,硅藻休眠体和异养型甲藻孢囊百分率均较高,而鱼类养殖区还出现了较高蓝藻孢子丰度.本次调查中还出现了易碎藻和艉杆藻等在本海域浮游植物调查中未记录种类的孢囊,有毒有害赤潮生物的休眠体如亚历山大藻、链状裸甲藻、旋沟藻、巴哈马梨甲藻孢囊等也在此次调查中频繁出现,说明大亚湾海域存在较大的有毒有害赤潮发生的风险.  相似文献   

20.
A modified version of the Coomassie Brilliant Blue dye binding protein assay has been developed for oceanographic samples and intercalibrated with the widely used Lowry assay. Particulate protein measurements were made at seven stations in the Gulf of Maine using the method. Measurements were made on cell-free homogenates. Protein concentration ranged from 2 to 212 μg l?1 (0.02–0.68 μg at N l?1) and averaged 58 μg l?1. ETS activity, chlorophyll and particulate nitrogen were significantly correlated with protein concentrations.  相似文献   

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