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1.
金属硫蛋白(Metallothionein,MT)是一类富含半胱氨酸的小分子蛋白质,参与机体重金属解毒和金属元素代谢等生理过程。本研究采用RACE技术,克隆获得了菲律宾蛤仔(Ruditapes philippinarum)金属硫蛋白(RpMT)的全长cDNA序列。RpMT的cDNA全长为570bp,编码75个氨基酸,包含15个MT所特有的Cys-Xn-Cys结构。采用实时荧光定量PCR技术,分析了两种壳色菲律宾蛤仔(白蛤和斑马蛤)RpMT基因在Cd2+暴露后的表达变化。结果发现:Cd2+急性和亚慢性暴露均可导致两种壳色蛤仔消化腺和鳃组织RpMT基因表达量的显著上调;暴露后两种壳色蛤仔鳃组织RpMT基因表达量的增加幅度均高于消化腺组织,且以白蛤鳃组织基因表达水平的上调幅度较高。上述结果表明,RpMT可能在菲律宾蛤仔抵御Cd2+胁迫过程中发挥了重要作用。  相似文献   

2.
对2018年1月在广东环雷州半岛近海海域采集的海底表层沉积物、鲬鱼(Platycephalus indicus)进行汞含量的测定,分析比较了鲬鱼不同部位汞含量的差异,并采用单因子污染指数法对沉积物和鲬鱼的汞污染状况进行了风险评价。结果表明,环雷州半岛海域表层沉积物中汞含量范围为0.005×10-6~0.359×10-6,平均值为0.081×10-6,雷州半岛东部海域表层沉积物汞含量高于南部和西部海域。鲬鱼的鳃中汞含量范围为0.032×10-6~0.034×10-6,平均值为0.033×10-6;肌肉中的汞含量范围为0.065×10-6~0.080×10-6,平均值为0.073×10-6;肝脏中的汞含量范围为0.228×10-6~0.270×10-6,平均值为0.249×10-6。鲬鱼样品的汞含量都呈现出肝脏>肌肉>鳃。单因子污染指数评价结果显示,雷州半岛海域鲬鱼受到汞的轻微污染,雷州半岛东部海域表层沉积物存在汞污染生态风险,而其南部、西部海域尚未受到汞污染的威胁。  相似文献   

3.
Biochemical characterization of cholinesterase activity (ChE) was carried out on the Antarctic scallop Adamussium colbecki collected in winter 2000 from Campo Icaro (Ross Sea, Antarctica) in order to increase its suitability as a sentinel organism for monitoring the Antarctic environment. The digestive gland, gills and adductor muscle were investigated for substrate specificity and inhibitors sensitivity using acetylthiocholine iodide (ASCh) and butyrylthiocholine iodide (BSCh) as substrates and tetra (monoisopropyl)pyrophosphor-tetramide (Iso-OMPA), 1,5-bis(4-allyldimethylammoniumphenyl)-penthan-3-one dibromide (BW284c51) and the insecticide chlorpyrifos as inhibitors. Effect of in vivo exposure to ZnCl2 was also investigated. All the tissues expressed ChE activity (gill > adductor muscle > digestive gland) and low substrates specificity throughout the hydrolysis of both ASCh and BSCh substrates. Partial (25–29%) and total inhibition (100%) of ChE activity in gills was demonstrated following in vitro incubation with Iso-OMPA and BW284c51 (3 mM), respectively. Concentration-dependent inhibition was also evident with chlorpyrifos in the range 10−4–10−10 M (IC50 10−6) while in vivo exposure to ZnCl2 did not seem to affect ChE activity in the scallop. The potential use of ChE in the A. colbecki as biomarker for monitoring water contamination in the marine Antarctic environment is discussed.  相似文献   

4.
张超  李永仁  郭永军  梁健 《海洋科学》2020,44(3):113-122
为研究港原油对毛蚶部分抗氧化酶和代谢酶的影响,设置0.01、0.1、1、3mg/L大港原油水溶液性成分(WSF),采用暴露法研究毛蚶天津群体的鳃、斧足中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GPX)抗氧化酶等抗氧化酶及酸性磷酸酶(ACP)、碱性磷酸酶(AKP)、钠/钾泵(Na+/K+ATPase)、钙泵(Ga2+/Mg2+ATPase)等代谢相关酶的活性变化,测定丙二醛(MDA)含量,采用整合生物标志物(Integratedbiomarkerresponse,IBR)进行分析。结果表明,毛蚶鳃和斧足中SOD、CAT、GPX表现出一定的剂量-效应关系, MDA含量呈先升高后降低的趋势, 3 mg/L组表现出MDA累积; ACP、AKP、Na+/K+ATPase、Ga2+/Mg2+ATPase表现出一定剂量-效应关系,酸性磷酸酶较碱性磷酸酶响应更迅速, Na+/K<...  相似文献   

5.
Metallothioneins and lysosomes are known to be involved in cellular detoxication and sequestration of certain metals1–3 and both have been identified in this role in elimination of copper from marine mussels (Mytilus edulis/galloprovincialis).3 Cadmium (Cd), however, has been shown to persist in the cells of the digestive gland for long periods with only minimal elimination. An experiment was designed to test the effects of Cd on the fragility of lysosomal membranes in the digestive cells as a measure of cellular injury,4,5 metallothionein content of the digestive gland and cadmium concentration in this organ. Phenanthrene was used also to destabilise lysosomal membranes6 in order to test if increased lysosomal fragility interfered with cadmium metabolism and detoxication. The results demonstrated that Cd induced metallothionein synthesis and that elimination of Cd was minimal after 28 days in clear seawater. Lysosomal fragility was initially increased but this effect was soon reversed, even with continued exposure to Cd. The lysosomal destabiliser, phenanthrene, did not appear to affect accumulation of Cd or levels of metallothionein.  相似文献   

6.
Our study investigates biomarker responses and survival of Macoma nasuta exposed to sediments collected from six locations in northern San Francisco Bay. Biomarkers analyzed were stress proteins (hsp70) in gill, mantle and digestive gland, lysosomal membrane damage and histopathologic lesions. Sediments and clam tissues were analyzed for a comprehensive suite of heavy metals and trace organic pollutants. Sediment grain size and organic carbon content were determined. Clams accumulated metals, polyaromatic hydrocarbons (PAHs) and organochlorine pesticides (aldrin and p,p-DDT and its metabolites p,p-DDD and p,p-DDE). Pearson and Spearman correlation analysis revealed that mortality, hsp70 in gill and histopathologic lesion scores in gonads, and lysosomal membrane damage were significantly correlated with tissue concentrations of DDT and/or its metabolites. Tissue concentrations of metals, in particular nickel, chromium, and copper, were associated with macrophage aggregates in digestive gland and germ cell necrosis. Cadmium was linked to mortality and lysosomal membrane damage.  相似文献   

7.
During a two-week period, Rangia rapidly accumulates mercury (as mercuric chloride) from dilute solution (0·03–0·05 ppm) with the majority being located in gill and mantle tissue. Initial depuration (5 h) in clean seawater is rapid in all tissues. From 10–192 h concentrations of mercury in gill, foot and adductor muscles remain stable while mantle, haemolymph and viscera levels continue to fluctuate in a manner suggesting internal redistribution of mercury. After eight days of depuration, the viscera contains the most mercury and appears to be the only tissue to increase its proportionate share of the total tissue mercury. After fifteen weeks of depuration, residual mercury in whole clams is about 20% of initial concentrations. Acute decreases in salinity (15% S to 2% S) enhance Rangia's ability to depurate mercury.  相似文献   

8.
Metals which induce the synthesis of metallothioneins, such as cadmium and mercury, also elevate glutathione (GSH) concentrations in fish tissues.1,2 To determine whether metals which do not stimulate metallothionein synthesis increase GSH levels, the GSH status of Atlantic croaker tissues was examined after oral administration of lead. Lead significantly increased GSH levels in the intestine and liver, but not in kidney or brain tissues. Lead was not bound to GSH and the increase in GSH did not appear to be linked to lead accumulation in these tissues. Investigations of hepatic GSH synthesis and turnover in vitro using a radiolabelled amino acid precursor, C14 glycine, showed that the elevation of GSH was due to an increase in GSH synthesis rather than decreased utilization of GSH. These studies suggest that the interactions between GSH and lead are indirect and involve a stimulatory effect of the metal on the activity of the enzymes for GSH synthesis.  相似文献   

9.
海水酸化暴露可对海洋生物产生多层面的影响。本研究以潮间带野生与潮下带养殖长牡蛎(不同生境背景)的不同组织(鳃、外套膜及消化腺)为研究对象,分析在室内调控p CO2模拟海水酸化暴露条件下,其基础代谢活动、能量代谢以及氧化应激相关指标的变化情况。结果显示:海水酸化暴露后,两种长牡蛎(Crassostrea gigas)的基础代谢过程均受到了一定抑制作用且受影响程度差异明显。潮间带野生与潮下带养殖长牡蛎的关键生理过程(能量代谢及氧化应激)对海水酸化暴露存在不同的响应变化,表明两种长牡蛎应对海水酸化的调节机制可能存在差异。依据PLS-DA分析结果显示,在所有生理指标中,对样本的差异贡献较高(VIP值>1)的指标为:SDH、AST、ATPase、ATP含量、糖原含量、CAT、GST及SOD,表明海水酸化暴露后,在两种长牡蛎的3种组织中上述指标的响应变化程度更大。综合评价分析多个生理指标的整体变化揭示:在海水酸化暴露条件下,潮间带野生长牡蛎比潮下带养殖长牡蛎对海水酸化的生理响应更为剧烈;相比于鳃及消化腺组织,长牡蛎外套膜组织可能受影响更大。  相似文献   

10.
The chemical speciation of dissolved mercury in surface waters of Galveston Bay was determined using the concentrations of mercury-complexing ligands and conditional stability constants of mercury-ligand complexes. Two classes of natural ligands associated with dissolved organic matter were determined by a competitive ligand exchange-solvent solvent extraction (CLE-SSE) method: a strong class (Ls), ranging from 19 to 93 pM with an average conditional stability constant (KHgLs) of 1028, and a weak class (Lw) ranging from 1.4 to 9.8 nM with an average KHgLs of 1023. The range of conditional stability constants between mercury and natural ligands suggested that sulfides and thiolates are important binding sites for dissolved mercury in estuarine waters. A positive correlation between the estuarine distribution of dissolved glutathione and that of mercury-complexing ligands supported this suggestion. Thermodynamic equilibrium modeling using stability constants for HgL, HgClx, Hg(OH)x, and HgCl(OH) and concentrations of each ligand demonstrated that almost all of the dissolved mercury (> 99%) in Galveston Bay was complexed by natural ligands associated with dissolved organic matter. The importance of low concentrations of high-affinity ligands that may originate in the biological system (i.e., glutathione and phytochelatin) suggests that the greater portion of bulk dissolved organic matter may not be important for mercury complexation in estuarine surface waters.  相似文献   

11.
为研究青蛤(Cyclina sinensis)对重金属Cu~(2+)的蓄积作用及免疫机能的影响,开展了Cu~(2+)对青蛤的急性毒性实验,观察青蛤在96 h Cu~(2+)半致死浓度和安全浓度胁迫下,不同组织的蓄积趋势,及血淋巴液中SOD、CAT和ACP活性的变化。结果显示:Cu~(2+)的半致死浓度为0.807 mg/L,安全浓度为0.00807 mg/L。鳃和内脏团组织中蓄积的Cu~(2+)浓度与处理时间呈正相关;在半致死和安全浓度胁迫下,鳃组织中Cu~(2+)蓄积速度均快于内脏团;在半致死浓度胁迫下血淋巴液中SOD、CAT和ACP活性呈现先诱导再抑制的趋势,安全浓度下无明显变化,但均高于对照组水平。  相似文献   

12.
构建了文蛤铁蛋白的重组表达质粒 pGEX-4T-1-MmeFer, 在大肠杆菌 Escherichia coli 中获得了重组蛋白, 经酶解和质谱鉴定该蛋白为重组文蛤铁蛋白(rGST-MmeFer), 切除 GST 标签的重组文蛤铁蛋白(rMmeFer)具有氧化和吸收铁离子的功能。利用 rGST-MmeFer 制备了兔抗文蛤铁蛋白的多克隆抗体, 进而利用 Western blot 的方法对文蛤铁蛋白的组织表达分布进行了研究。结果表明, 铁蛋白在文蛤各组织中均有分布, 其中消化腺中表达量最高, 足中表达量最低。此外, 利用实时定量PCR 分析了文蛤消化腺、外套膜和鳃组织中铁蛋白 mRNA 的表达, 发现铁蛋白 mRNA 在外套膜中表达量最高, 消化腺次之, 鳃中表达量最低, 差异达到显著水平(P<0.05)。上述结果为进一步研究铁蛋白参与贝类贝壳形成的分子机制奠定了基础。  相似文献   

13.
The Mussel Watch program conducted along the French coasts for the last 20 years indicates that the highest mercury concentrations in the soft tissue of the blue mussel (Mytilus edulis) occur in animals from the eastern part of Seine Bay on the south coast of the English Channel, the “Pays de Caux”. This region is characterized by the presence of intertidal and submarine groundwater discharges, and no particular mercury effluent has been reported in its vicinity. Two groundwater emergence systems in the karstic coastal zone of the Pays de Caux (Etretat and Yport with slow and fast water percolation pathways respectively) were seasonally sampled to study mercury distribution, partitioning and speciation in water. Samples were also collected in the freshwater–seawater mixing zones in order to compare mercury concentrations and speciation between these “subterranean” or “groundwater” estuaries and the adjacent macrotidal Seine estuary, characterized by a high turbidity zone (HTZ). The mercury concentrations in the soft tissue of mussels from the same areas were monitored at the same time.The means of the “dissolved” (< 0.45 μm) mercury concentrations (HgTD) in the groundwater springs were 0.99 ± 0.15 ng l− 1 (n = 18) and 0.44 ± 0.17 ng l− 1 (n = 17) at Etretat and Yport respectively. High HgTD concentrations were associated with strong runoff over short water pathways during storm periods, while low concentrations were associated with long groundwater pathways. Mean particulate mercury concentrations were 0.22 ± 0.05 ng mg− 1 (n = 16) and 0.16 ± 0.10 ng mg− 1 (n = 17) at Etretat and Yport respectively, and decreased with increasing particle concentration probably as a result of dilution by particles from soil erosion. Groundwater mercury speciation was characterized by high reactive-to-total mercury ratios in the dissolved phase (HgRD/HgTD: 44–95%), and very low total monomethylmercury concentrations (MMHg < 8 pg l− 1). The HgTD distributions in the Yport and Etretat mixing zones were similar (overall mean concentration of 0.73 ± 0.21 ng l− 1, n = 43), but higher than those measured in the adjacent industrialized Seine estuary (mean: 0.31 ± 0.11 ng l− 1, n = 67). In the coastal waters along the Pays de Caux dissolved monomethylmercury (MMHgD) concentrations varied from 9.5 to 13.5 pg l− 1 (2 to 8% of the HgTD). Comparable levels were measured in the Seine estuary (range: 12.2– 21.1 pg l−1; 6–12% of the HgTD). These groundwater karstic estuaries seem to be mostly characterized by the higher HgTD and HgRD concentrations than in the adjacent HTZ Seine estuary. While the HTZ of the Seine estuary acts as a dissolved mercury removal system, the low turbid mixing zone of the Pays de Caux receives the dissolved mercury inputs from the groundwater seepage with an apparent Hg transfer from the particulate phase to the “dissolved” phase (< 0.45 μm). In parallel, the soft tissue of mussels collected near the groundwater discharges, at Etretat and Yport, exhibited significantly higher values than those found in the mussel from the mouth of the Seine estuary. We observe that this difference mimics the differences found in the mercury distribution in the water, and argue that the dissolved phase of the groundwater estuaries and coastal particles are significant sources of bioavailable mercury for mussels.  相似文献   

14.
螯虾是重要的水产养殖动物,也是研究甲壳动物疾病与免疫的一个良好模型.在研究不同组织的功能及受病原感染的影响时,需要了解组织内的细胞数量.但是,目前常用的细胞计数方法仅适用于单细胞悬液,无法用于实体组织.本研究以红螯螯虾( Cherax quadricarinatus)为对象,建立了基于基因拷贝数定量分析的实体组织细胞数...  相似文献   

15.
利用构建的cDNA文库及高通量测序方法,获得金属硫蛋白及硫氧还蛋白基因全长。结果表明,青蛤金属硫蛋白和硫氧还蛋白的cDNA全长分别为776bp和804bp,金属硫蛋白基因编码74个氨基酸,包含7个CXC特征结构;硫氧还蛋白基因编码165氨基酸,包含5个氨基酸构成的活性中心。采用实时定量PCR方法分析了两种基因在Cd2+胁迫下的表达变化,结果显示,金属硫蛋白基因在Cd2+胁迫下6—12hmRNA表达量急剧上升,与对照组有显著性差异(P<0.01)。硫氧还蛋白基因在Cd2+胁迫下mRNA在6h明显升高,在6—24h与对照组出现显著性差异(P<0.05)。说明Cd2+能够诱导青蛤金属硫蛋白和硫氧还蛋白基因表达的时序性升高,二者的转录过程反映了贝类抵御重金属胁迫的分子调控过程。  相似文献   

16.
雌雄波纹巴非蛤不同组织中总类胡萝卜素含量比较分析   总被引:1,自引:0,他引:1  
本文分析了波纹巴非蛤(Paphia undulata)斧足、外套膜、鳃、闭壳肌、性腺5个组织以及不同性别的总类胡萝卜素含量(TCC)。结果显示:波纹巴非蛤5个组织的总类胡萝卜素含量除外套膜与鳃之间没有显著性差异外(P0.05),其他各组织之间差异均达到极显著水平(P0.01),各组织总类胡萝卜素含量从高到低依次为:斧足性腺外套膜鳃闭壳肌;不同性别波纹巴非蛤的斧足、外套膜、鳃、闭壳肌中总类胡萝卜素的含量没有显著性差异(P0.05),但精巢中总类胡萝卜素的含量要显著高于卵巢(P0.05)。本研究结果初步揭示了波纹巴非蛤体内总类胡萝卜素含量的分布规律。  相似文献   

17.
Investigations on seasonal variation in oxidative stress biomarkers were carried out on the natural population of green-lipped mussel Perna viridis collected from Bambolim beach area of Goa. Oxidative stress indices such as lipid peroxidation (LPX), hydrogen peroxide (H2O2), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX), glutathione transferase (GST), glutathione reductase (GR), reduced glutathione (GSH) and ascorbic acid (ASA) were measured in gills and digestive gland of P. viridis during February, May, August and November. The present study reveals two important aspects regarding the antioxidant defence status of tissues of P. viridis. Firstly, antioxidant capacity of tissues of P. viridis exhibits seasonal variation. Secondly, various components of antioxidant capacity such as oxidative stress markers, levels of antioxidant enzymes and small antioxidant molecules vary differently in tissues with respect to different seasons. Although the oxidative stress status of gills and digestive gland of P. viridis expressed in terms of LPX and H2O2 was the lowest in February, its level was maximal in gills and digestive gland during May and November, respectively. While activities of SOD and GPX of tissues of P. viridis were found to be low in August, activities of CAT and GR were recorded to be low in February. GST activity in gills although remained high in February, in digestive gland elevated values were recorded in August and November. A seasonal variation in the levels of small antioxidant molecules was also noticed. Among non enzymatic antioxidants ASA content of tissues was maximal in May and August in comparison to February and November, but GSH remained high in November. It therefore appears that environmental factors may play a crucial role in regulating the oxidative stress capacity of tissues of P. viridis.  相似文献   

18.
克隆得到缢蛏天然抗性相关巨噬蛋白2(Sc-Nramp2)基因的cDNA全长序列3 681 bp,该基因的开放阅读框有1 776 bp,编码591个氨基酸,预测分子量为65.86 kDa;其结构具有Slc11蛋白家族的典型特征,包括有10个典型的跨膜结构和2个糖基化位点。Sc-Nramp2基因3'-UTR有2个类似于脊椎动物Nramp2中铁反应控制蛋白结合位点;同源性分析表明,Sc-Nramp2和太平洋牡蛎Nramp2-like的同源性最高为71.6%。实时荧光定量PCR结果表明,Sc-Nramp2基因在闭壳肌、外套膜、肝胰腺、斧足、水管和鳃6个组织中均有表达,其中肝胰腺中的表达量最高,其次是鳃,与其他组织均有极显著性差异(P<0.01);注射副溶血弧菌后,肝胰腺中Sc-Nramp2基因的表达量较对照组显著上调(P<0.01),且表达量呈现先上升后下降的趋势,在12 h时达到最大表达量,推测Sc-Nramp2基因参与了缢蛏非特异性免疫应答反应。  相似文献   

19.
为探明舟山海域网箱养殖小黄鱼(Larimichthys polyactis)暴发的以体表溃疡、眼球溃烂、内脏出现白色结节为主要症状的疾病病因, 对患病小黄鱼体表溃疡处和结节病灶处采集组织样本, 在血琼脂平板上划线培养, 通过分离、纯化到同一优势菌株ZHNK2101。经形态学观察、生理生化鉴定、16S rRNA基因片段扩增及序列比对,确定该菌株为鰤鱼诺卡氏菌(Nocardia seriolae)。药敏试验结果显示, 菌株ZHNK2101对哌拉西林、头孢曲松、丁胺卡那、庆大霉素等16种抗生素敏感。回归感染试验表明, 该菌株可以感染小黄鱼并引起和自然感染相同的症状, 半致死浓度LD50为7.28×104 CFU/尾。另外,该病株会引起小黄鱼鳃、肝、肾和脾脏发生病理变化, 主要表现为鳃丝紊乱,肝脏、肾脏、脾脏组织细胞纤维化形成病理性结节。试验首次报道了小黄鱼感染鰤鱼诺卡氏菌的病例, 并从理化特性、药敏试验、回归感染及组织病理变化等方面对该菌株的特性进行了初步研究, 为小黄鱼养殖过程中由鰤鱼诺卡氏菌引起的内脏白点病的早期预防和治疗提供基础数据。  相似文献   

20.
Catalase is an important antioxidant protein that can protect organisms against various forms of oxidative damage by eliminating hydrogen peroxide. In this study, the catalase c DNA of Paphia textile(Pt CAT) was cloned using RTPCR and rapid amplification of c DNA ends(RACE). Pt CAT is 1 921 bp long and consists of a 5′-UTR of 50 bp, a 3′-UTR of 349 bp, and an ORF of 1 542 bp that encodes 513 amino acids with a molecular weight of 58.4 k D and an estimated isoelectric point of 8.2. Sequence alignment indicated that Pt CAT contained a highly conserved catalytic signature motif(~(61)FNRERIPERVVHAKGAG~(77)), a proximal heme-ligand signature sequence(~(352)RLFSYSDP~(359)), and three catalytic amino acid residues(H~(72), N~(145), and Y~(356)). Pt CAT also contains two putative N-glycosylation sites(~(34)NKT~(36) and ~(437)NFT~(439)) and a peroxisome-targeting signal(~(511)AQL~(513)). Furthermore, Pt CAT shares 53%–88% identity and 29%–89% similarity with other catalase amino acid sequences. Pt CAT m RNA was present in all tested organs, including the heart, digestive gland, adductor muscle, gonad, gill, and mantle, but its expression was highest in the digestive gland. High-temperature-induced stress produced two expression patterns of Pt CAT m RNA: first, an initial up-regulation followed by a down-regulation in the heart, digestive gland, and gonad and, second, consistent down-regulation in all other organs. These results demonstrate that Pt CAT is a typical member of the catalase family and might be involved in the responses to harmful environmental factors.  相似文献   

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