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1.
扇贝中麻痹性毒素的提取与分离纯化   总被引:1,自引:1,他引:0  
为了获得麻痹性毒素标准品,以毒化扇贝为试验材料,对扇贝中的麻痹性毒素(PSP毒素)进行了提取和分离纯化。以酸性80%乙醇溶液反复提取麻痹性毒素,得到了总毒性为6170MU的毒素粗提液。毒素粗提液经过超滤、Bio-GelP-2凝胶柱层析和Bio-Rex70离子交换柱层析等二次层析柱分离纯化后得到以膝沟藻毒素群(GTXs)为主的纯化PSP毒素。HPLC分析结果显示,扇贝中主要含有膝沟藻毒素GTX3,GTX1、GTX2、GTX3、和GTX4的组成比例约为2∶4∶14∶1(以HPLC上的峰面积比例计)。纯化后的毒素可以作为标准物质用于GTX1-4的HPLC分析。  相似文献   

2.
将栉孔扇贝(Chlamys farreri)直接暴露于产毒藻,通过比较各组织器官中毒素的蓄积及代谢转化特征,研究麻痹性贝类毒素(Paralytic Shellfish Toxins,PSTs)在栉孔扇贝体内危害形成的过程。结果显示,分布于中国的一株塔玛亚历山大藻(Alexandrium tamarense,AT5-3株),其生长及产毒性状稳定,产毒能力强,主要成分为Gonyautoxins-1-4(GTX14),单细胞产毒能力为7.95 fmol/cell;栉孔扇贝对该产毒藻具有较强摄食能力及PSTs蓄积能力,总体蓄积效率达到84.4%,最高蓄积浓度为1903μg STX eq./kg,不同组织蓄积能力由高到低依次为内脏团、性腺、外套膜、闭壳肌;内脏团对PSTs的代谢能力也最强,是该毒素在栉孔扇贝体内蓄积代谢的靶器官;此外,栉孔扇贝对PSTs表现出较强的生物转化能力,主要转化途径为:N-磺酰胺甲酰基类毒素(N-Sulfocarbamoylgonyautoxin-2,C1)→膝沟藻毒素(Gonyautxins-2,GTX2)/脱氨甲酰基类毒素(Decarbamoylgonyautoxins-2,dc GTX2),N-磺酰胺甲酰基类毒素(N-Sulfocarbamoylgonyautoxin-3,C2)→膝沟藻毒素(Gonyautxins-3,GTX3)。本研究中,栉孔扇贝对PSTs总体呈现出迅速蓄积和缓慢代谢的特点,同时,GTX14和NEO等高毒成分所占比例较高,造成扇贝中PSTs毒性和风险性均显著升高。本研究有助于科学评估PSTs危害的形成机制,为系统监控我国PSTs风险性提供科学支撑。  相似文献   

3.
为获知牡蛎对有毒微小亚历山大藻(A. minutum)的滤食以及蓄积PSP毒素的规律性,本实验采用细胞毒性为93.42±2.55×10-6 MU/cell的A. minutum藻投喂牡蛎,以无毒扁藻投喂作对照,首先研究了0.5-3.0×103 cells/mL6个不同浓度的毒藻对牡蛎滤食的影响,然后投喂浓度为1.5×103 cells/mL的毒藻,分别研究15 h短期蓄积和5天长期蓄积对牡蛎体内毒素的影响及规律。结果表明,在0.5~1.5×103 cells/mL毒藻浓度范围内牡蛎清滤率较高,超过1.5×103 cells/mL时牡蛎清滤率显著下降(P<0.01),由此确定后期投喂毒藻的浓度“阀值”为1.5×103 cells/mL;短期蓄积实验牡蛎在0~6 h内将有毒藻滤食尽,其体内PSP毒素在6~12 h间达到最大值149.6±10.5 MU/100 g,随后开始下降。长期蓄积实验过程中,牡蛎的清滤率没有发生显著变化(P>0.01),在蓄积实验的第2天牡蛎体内PSP毒素水平超过国家限量标准(400 MU/100g),在5天实验结束时,牡蛎体内PSP毒素水平高达3069.2±178.2 MU/100 g,高于国家限量标准的7.7倍。实验结果作为PSP在牡蛎中蓄积及代谢的重要基础数据,为研究PSP的脱除及净化提供思路。  相似文献   

4.
王亚军  余新威  方力  刘其林  任斌  罗红宇 《水产学报》2017,41(10):1588-1598
为考察营养成分以及培养基础液再利用对一株微小亚历山大藻的生长和产毒的影响,采用单因素实验分别研究了氮(NaNO_3)、磷(NaH_2PO_4)、微量元素(FeCl_3、Na2EDTA、CuSO_4、Na_2MoO_4、ZnSO_4、CoCl_2、MnCl_2)、维生素(Vitamin B12、Vitamin H、Vitamin B1)、碳(NaHCO_3)的不同含量以及海水(培养基础液)利用方式(不循环与循环利用)对微小亚历山大藻C4生长、毒素含量(μmol/L)与毒素组成的影响。结果显示,氮、磷浓度对C4藻的毒素总含量(μmol/L)有显著影响,其他因素影响均不显著;氮、磷、碳及微量元素的浓度对GTX1/4(GTX1+GTX4)在总毒素(GTX1+GTX2+GTX3+GTX4)中所占比例均影响显著,而维生素浓度和海水循环利用对GTX1/4所占比例影响均不明显。氮浓度在0~883μmol/L范围内,毒素含量与氮浓度呈正相关关系,氮浓度进一步增加,毒素含量没有明显变化;磷浓度在0~145.2μmol/L范围内增加,毒素含量先增后降,最终保持稳定的趋势。最佳的产毒条件为氮883μmol/L,磷18.15μmol/L,微量元素为f/2海水培养基中微量元素的0.5倍,碳不添加。  相似文献   

5.
为监测渤海海域唐山贝类养殖区贝类毒素的污染情况,防止食用贝类中毒事件发生,于2019年10月—2020年9月间,每月持续在渤海海域唐山贝类养殖区采集四角蛤(Mactra veneriformis)、菲律宾蛤仔(Ruditapes philippinarum)、脉红螺(Rapana venosa)、牡蛎(Crassostrea gigas)、青蛤(Cyclina sinensis)、文蛤(Meretrix meretrix)和硬壳蛤(Mercenaria mercenaria) 7种经济贝类样品,采用高效液相色谱–串联质谱(HPLC-MS/MS)法测试了5种腹泻性贝类毒素(diarrhetic shellfish poisoning, DSP)和14种麻痹性贝类毒素(paralytic shellfish poisoning, PSP)。结果显示,在7种经济贝类样品中均未检出DSP。检出的PSP成分包括石房蛤毒素(Saxitoxin, STX)、膝沟藻毒素1 (Gonyautoxin 1, GTX 1)、膝沟藻毒素2 (Gonyautoxin 2, GTX 2)和脱氨甲酰基膝沟藻毒素3 (Ddecarbamoy l gonyautoxin 3, dcGTX 3),其中,GTX 1含量最高且最高值为537.95 μg/kg。不同季节贝类毒素蓄积含量有一定差异,PSP主要集中在4月检出。菲律宾蛤仔、牡蛎、文蛤和硬壳蛤中PSP的检出率分别为11.76%、47.06%、5.90%和8.82%,其他贝类均未检出。PSP总量均低于欧盟及中国的食用安全限量标准800 μg STXeq/kg。应用风险熵值法和点评估法进行食用安全风险评估,显示风险熵值和暴露风险指数均在安全范围内,结果表明,渤海海域唐山贝类养殖区7种经济贝类不存在食用安全风险。  相似文献   

6.
氮杂螺环酸毒素(azaspiracids,AZAs)隶属八大类海洋贝类毒素,为一类含氮且具独特螺环结构的聚醚类毒素。采用液相色谱-三重四级杆复合离子阱质谱,研究了分离自中国近海的1株氮杂螺环酸产毒藻腹孔环胺藻(Azadinium poporum,AZDY06)所产氮杂螺环酸毒素在栉孔扇贝体内的蓄积、分布和生物转化机制。通过将栉孔扇贝暴露产毒藻的方式,分析扇贝内脏团、裙边、闭壳肌和其他可食组织4个组织部位的AZAs及其代谢产物分布,研究栉孔扇贝对毒素的代谢机理。结果表明,AZDY06主要产生AZA2毒素,单细胞产毒能力最高为(7.05±0.52)fg/cell;扇贝12 h内摄食5×10~7个产毒藻细胞后,体内AZAs毒素含量已超欧盟安全限量,达165.3μg AZA1eq/kg,蓄积效率为78.2%。AZAs毒素在扇贝各组织间分布存在显著差异:内脏团其他可食组织外套膜闭壳肌。AZA2在扇贝中潜在转化方式为羟基化、去羧基化和氧化,共生成4种代谢产物:AZA6、AZA12、AZA19和AZA23,其中AZA19为最主要代谢产物,约占总毒素40%,其他代谢产物含量较低。本研究证明中国近海分布氮杂螺环酸产毒藻毒性危害较强,建议有关部门加快制定AZAs限量标准。  相似文献   

7.
使用液相色谱串联质谱法(LC-MS/MS)检测9种天津市售主要经济贝类中13种麻痹性贝类毒素(paralytic shellfish poisoning,PSP)的含量,分析PSP的污染现状和成分特征。结果显示,共计6个品种(紫贻贝、虾夷扇贝、牡蛎、毛蚶、脉红螺、菲律宾蛤仔)检出PSP。其中虾夷扇贝、紫贻贝、毛蚶3个品种的检出率较高且含量较高,检出率分别为66.7%、58.3%、58.3%。PSP中检出率较高的毒素包括N-磺酰胺甲酰基类毒素C1、C2和膝沟藻毒素GTX1、GTX2、GTX3,毒力占比主要以膝沟藻毒素GTX1-4为主,毛蚶蓄积STX毒素的能力强于其他贝类。全年的PSP含量季节变化为夏季最高,冬季最低。PSP主要存在于贝类的消化腺内,全年不同月份的PSP含量均在安全食用范围内。  相似文献   

8.
贝类毒素标准样品的制备是保障贝类质量安全的基础,本研究以有毒贻贝(Mytilus edulis)为原料,探究麻痹性贝类毒素标准样品的制备技术。以1%乙酸溶液提取贻贝中的麻痹性贝类毒素,采用羟丙基葡聚糖凝胶柱(Sephardex LH20)净化后收集目标馏分,再经高效液相色谱以TSK-GEL Hillic Amide-80色谱柱(4.6 mm×250.0 mm,5.0μm)分离、纯化,根据麻痹性贝类毒素的保留时间收集色谱峰流出液,连续制备合并馏分,40℃减压蒸馏浓缩近干,用75%乙腈水(含0.25%甲酸)溶解,即制得毒素标准样品溶液。分别采用四极杆/离子阱复合质谱与标准物质的多级质谱信息进行匹配,以高分辨质谱获得的精确质量数进行定性确证,样品溶液所含的麻痹性贝类毒素为4种膝沟藻毒素,分别为膝沟藻毒素-1-4 (Gonyautxins-1-4, GTX14)和膝沟藻毒素-2-3(Gonyautxins-2-3, GTX23)。均匀性实验中,经F检验表明在95%的置信区间范围内样品均匀性良好。稳定性考察实验中,标准样品在4℃和-18℃两种环境条件下储存6个月稳定性较好。标准样品经8家实验室联合定值,并评定了定值结果的不确定度。本研究制得的麻痹性贝类毒素标准样品为GTX 1(3 663μg/L)GTX4(1 147μg/L)和GTX 2(1 884μg/L)GTX3(797μg/L)的混合溶液,溶剂为75%乙腈水(含0.25%甲酸),为国内贝类毒素标准样品研发提供了技术支撑。[中国渔业质量与标准,2019,9(2):62-70]  相似文献   

9.
本研究将紫贻贝(Mytilus galloprovincialis)暴露于一株麻痹性贝类毒素(paralytic shellfish toxins, PSTs)优势产毒藻——链状亚历山大藻(Alexandrium catenella, GY-H25株),模拟现场赤潮藻密度,探究了紫贻贝内脏团和可食组织中蓄积代谢及生物转化过程,并通过蓄积代谢动力学,重点比较了不同细胞密度GY-H25对紫贻贝体内毒素蓄积代谢和转化情况的影响。结果显示,GY-H25生长及产毒稳定,PSTs组分主要为N-磺酰胺甲酰基类毒素(C1和C2),单细胞最高产毒能力为2.96 pg STXeq/cell。暴露实验中,紫贻贝对PSTs有较强的蓄积作用,2种暴露浓度下PSTs含量变化趋势一致,实验结束时,2种暴露组紫贻贝内脏团中PSTs均超过欧盟国际限量标准(800 μg STXeq/kg),但可食组织则均低于限量标准;比较发现,高浓度组紫贻贝内脏团最高蓄积浓度达到6 815.36 μg STXeq/kg,且高浓度组暴露期间平均蓄积速率为17.89%,显著高于低浓度组13.06%的蓄积速率。另外,紫贻贝对PSTs表现出较强的生物转化能力,在对C1、C2和GTX5三者的转化研究中发现,快速代谢时期和平稳期C2→GTX5的转化为GTX5生成的主要途径,同时期C1的相关转化中C1→GTX5途径超过C2→C1,致使C1整体占比减少。综合评估紫贻贝中PSTs转化产物和毒性当量因子(toxic equivalency factor, TEF),发现紫贻贝对PSTs代谢转化进一步促使高毒性GTX5的生成和占比提升,总体终端毒性升高,这也可能是秦皇岛紫贻贝中PSTs风险严峻的主要原因之一。因此,本研究有助于科学评估紫贻贝中PSTs风险,为建立区域性风险监测技术提供科学基础。  相似文献   

10.
《海洋渔业》2021,43(3)
为探究塔玛亚历山大藻(Alexandrium tamarense)产麻痹性贝类毒素(paralytic shellfish poisoning, PSP)的能力,通过设置不同空间效应、盐度和营养素浓度对塔玛亚历山大藻进行培养,分析不同培养条件下塔玛亚历山大藻产麻痹性贝类毒素的能力。结果表明:1)塔玛亚历山大藻中至少含有13种PSP组分,其中主要成分为GTX1、GTX4、GTX5和C1、C2,占总毒素的97.2%。当藻细胞数量低于4×10~3个时,13种组分均不能定量检出;当藻细胞数量在4×10~3~5.9×10~6个时,13种组分不能完全定量检出;当藻细胞数高于5.9×10~6个时,13种组分均能够检出。2)不同条件下塔玛亚历山大藻产PSP能力比较,盐度25、35培养组显著高于盐度15培养组,1 L培养组显著高于0.1 L培养组,寡营养培养组显著高于富营养组。研究结果表明,塔玛亚历山大藻具有丰富的毒素谱,且分别在盐度25、1 L培养液和1/4倍营养素浓度下具有最佳产毒能力。  相似文献   

11.
ABSTRACT: Paralytic toxicity was detected by tetrodotoxin (TTX) bioassay in four and five specimens of new toxic xanthid crab Xanthia lividus collected from Lanyu, south-east Taiwan in April 1999 and Hsiaoliuchiu south-west Taiwan in November 1999, respectively. The average toxicity of crab specimens collected from Lanyu and Hsiaoliuchiu was 230 ± 94 (mean ± SD) mouse units (MU) and 241 ± 114 MU, respectively. The toxin was partially purified by YM-1 membrane ultrafiltration and Bio-Gel P-2 column chromatography. Electrophoresis, high-performance liquid chromatography and gas chromatography–mass spectroscopy analyses showed that the crab toxin was composed mainly of tetrodotoxin (83%), as well as a small amount of gonyautoxin 1-4 (17%).  相似文献   

12.
李磊  蒋玫  王云龙 《海洋渔业》2019,41(3):346-353
邻苯二甲酸二丁酯(dibutyl phthalate,DBP)和邻苯二甲酸二辛酯(dinoctyl phthalate,DOP)是一类重要的工业用有机化合物,在水环境中广泛分布,对水生生物具有一定的毒性效应。为了研究DBP和DOP对大黄鱼(Larimichthys crocea)受精卵及初孵仔鱼的急性毒性效应,开展大黄鱼幼鱼在DBP和DOP中的暴露实验。结果显示,随着DBP和DOP浓度的升高,大黄鱼受精卵孵化率呈现逐步降低的趋势,死亡率、畸形率则呈现逐步升高的趋势。DBP和DOP暴露对大黄鱼初孵仔鱼均产生一定的致死效应,DBP和DOP对大黄鱼仔鱼的96 h半致死浓度分别为5.23 mg·L^-1 和6.57 mg·L^-1 。结果表明,DBP和DOP对大黄鱼鱼卵及初孵仔鱼均具有高毒毒性。DBP和DOP对水生生物的慢性毒性需要进一步关注。  相似文献   

13.
Tetrodotoxin (TTX) was detected by HPLC and LC-MS/MS in specimens of the horseshoe crab Carcinoscorpius rotundicauda collected from Tan Hai village, Vung Tau province, Vietnam, where cases of poisoning due to the consumption of horseshoe crab have frequently been reported. The soft tissue from 10 out of the 12 specimens analyzed showed considerable levels of TTX, making them unsafe for human consumption (81.2 ± 50.3 MU/g). Paralytic shellfish poisoning (PSP) toxins were also detected in all of the specimens, but the levels were low. These findings show that the frequency of occurrence of C. rotundicauda specimens with a high level of TTX is significantly high, and that the TTX is responsible for the food poisonings caused by the consumption of C. rotundicauda in this area.  相似文献   

14.
Dao  Ha Viet  Takata  Yoshinobu  Omura  Takuo  Sato  Shigeru  Fukuyo  Yasuwo  Kodama  Masaaki 《Fisheries Science》2009,75(2):507-512
Previously we reported that a significant level of domoic acid, a causative toxin for amnesic shellfish poisoning, was detected in bivalves belonging to a genus Spondylus from various tropical Asian countries. These findings indicate that causative plankton species for domoic acid widely distribute in these areas. In the present study, we monitored seasonal change of domoic acid level of Spondylus versicolor in association with that of plankton samples in Nha Phu Bay, Khanh Hoa, Vietnam from December 2004 to October 2005. The toxicity of S. versicolor showed distinct seasonal variation. During the period when domoic acid level of S. versicolor was increasing, a significant level of domoic acid was detected in the plankton samples, showing the correlation between these two parameters. These findings show that plankton causative for domoic acid occurred in the bay, and S. versicolor accumulate domoic acid during a bloom of the toxic plankton by food web transfer.  相似文献   

15.
The accumulation of paralytic shellfish poisoning (PSP) toxin in the crab Charybdis japonica was investigated in Kure Bay, when a bivalve Mytilus galloprovincialis, in the same water, showed toxicity caused by PSP toxin. In 2005, 17 specimens of the crab had PSP toxin over 4 MU/g in the hepatopancreas, and the highest toxicity was at 37.4 MU/g. Since the regulation limit of PSP for crab hepatopancreas was set in 2004, this is the first observation of regulatory level of PSP toxin in C. japonica. Unlike in the hepatopancreas of the crab, the hazardous level of the toxin was not detected in the muscle of the cephalothorax and the appendage. The toxin accumulation in the crab was also investigated by feeding toxic mussels to the crab. The crab retained the toxin mainly in the hepatopancreas, and the ratio of retention in the crab was from 12.9 to 24.6%. The toxin profiles, shown in the feeding experiments, suggest that the transformation of PSP toxin occurs in the crab because dcGTX2 and dcGTX3 was detected in all crab specimens despite the lack of these analogs in the mussels used as feed.  相似文献   

16.
During ecological speciation, diverging taxa have the potential to remain in close spatial proximity (i.e., sympatry or micro‐allopatry) theoretically allowing for continued contact and gene flow. In a system where incipient speciation of populations of the Atlantic molly (Poecilia mexicana) appears to be driven by abiotic factors, we investigated whether one of these factors, hydrogen sulphide (H2S) toxicity, also constitutes an effective barrier to slow migration within and between habitats. We addressed this experimentally by translocating individuals from high toxicity to lower toxicity within a toxic cave and by translocating individuals from the toxic cave to a nontoxic surface habitat. Using a stepwise‐backwards Cox regression, we found that overall mortality was low, but statistically significant mortality occurred when individuals were transferred from higher toxicity to lower toxicity. In addition, only males were negatively affected by being transferred from low levels of toxicity to nontoxic, surface waters. This indicates that in addition to abiotic factors, other mechanisms, like predation and sexual selection, must be important in maintaining population separation.  相似文献   

17.
李冰  张木子  黎明  袁莉霞  王日昕 《水产学报》2018,42(12):1889-1895
为了研究急性氨氮胁迫对黄颡鱼头肾巨噬细胞抗氧化及炎症相关基因表达的影响,实验通过建立黄颡鱼离体头肾细胞模型,开展为期96 h的急性氨氮胁迫实验。结果显示,0.14和0.28 mg/L总氨氮处理组SOD基因的相对表达量显著低于对照组,但0.28 mg/L总氨氮处理组GPX基因的相对表达量显著高于其他组;0.14和0.28 mg/L总氨氮处理组IL-1和TNF基因的相对表达量显著低于对照组,而0.28 mg/L总氨氮处理组IL-8基因的相对表达量显著低于0.14 mg/L总氨氮处理组和对照组;吖啶橙染色检测发现,0.14 mg/L总氨氮处理组实验鱼头肾巨噬细胞内可见边缘不规则的黄绿色荧光,0.28 mg/L总氨氮处理组实验鱼头肾巨噬细胞内可见致密浓染的黄绿色荧光信号。研究表明,氨氮毒性能够影响黄颡鱼头肾巨噬细胞抗氧化相关基因的表达;细胞凋亡可能是造成鱼类氨中毒致死的主要原因之一。  相似文献   

18.
研究了吡喹酮不同给药方式对鲫(Carassius auratus)的急性和亚急性毒性试验。急性毒性试验结果显示:浸泡时吡喹酮对鲫的24、48、96 h的半致死浓度(LC50)分别为54.80 mg/L、51.07 mg/L、50.78 mg/L;按每千克鱼体重10 g吡喹酮的剂量灌胃时,96 h未见明显中毒症状。亚急性试验结果显示:在不同的给药方式下,与对照组相比,低浓度的吡喹酮未对碱性磷酸酶(ALP)、超氧化物歧化酶(SOD)和谷丙转氨酶(ALT)的活性造成显著影响(P>0.05);随着试验时间的延长,较高浓度的吡喹酮使SOD活性先升高后降低,ALT活性显著(P<0.05)或极显著高于对照组(P<0.01)。  相似文献   

19.
ABSTRACT:     The difference in paralytic shellfish poisoning (PSP) toxin accumulation in the crabs Telmessus acutidens and Charybdis japonica was investigated at Onahama, Fukushima Prefecture, from 2002 to 2005. The level of toxin accumulation in the hepatopancreas of T. acutidens corresponded to that of mussels when examined on a yearly basis. In 2003, some crabs had a high toxicity of approximately 1000 MU, which compares to one-third of the human minimum lethal dose. Therefore T. acutidens should be monitored as a vector species of PSP toxin. The toxin profile of T. acutidens was also investigated. Because an increase in highly toxic species of PSP toxins with a decrease in low toxic species, such as N -sulfocarbamoyl-11-hydroxysulfate toxins, was not clearly observed between consecutive samples, toxin transformation in T. acutidens was considered to have a minimal impact on toxicity. PSP toxins were also detected in several specimens of C. japonica , but the highest toxicity was only 7.4 MU/g in the hepatopancreas. Because C. japonica is widely distributed in the coastal waters of Japan, investigation of toxin accumulation in the crab should be continued, including its investigation in different areas of water from Onahama, where the causative dinoflagellates of PSP occur in different conditions.  相似文献   

20.
Chloramphenicol (CAP) residues in carp (Cyprinus carpio L.) were investigated after intramuscular injection of a single-dose of 80 mg/kgbw. CAP concentrations in carp serum and muscle with and without addition of β-glucuronidase were determined by high-performance liquid chromatography (HPLC) with UV detection. The results showed that CAP-glucuronide (CAP-G), an important conjugate of CAP in the metabolic process and potentially toxic to human health, was observed after the fish were injected with CAP. CAP concentrations in serum and muscle with added β-glucuronidase were significantly (< 0.05) higher than those without added enzyme. For example, within 24 h of injection, average CAP concentrations in serum and muscle with enzyme digestion were 30 and 48% more than those without the enzyme. In order to exactly determine the trace CAP content of fish, for evaluation of food safety, it is suggested that samples be hydrolyzed with enzyme (e.g., β-glucuronidase).  相似文献   

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