Identification of heavy metal-contaminated Tegillarca granosa using laser-induced breakdown spectroscopy and linear regression for classification

Tegillarca granosa (T. granosa) is susceptible to heavy metals, which may pose a threat to consumer health. Thus, healthy and polluted T. granosa should be distinguished quickly. This study aimed to rapidly identify heavy metal pollution by using laser-induced breakdown spectroscopy (LIBS) coupled with linear regression classification (LRC). Five types of T. granosa were studied, namely, Cd-, Zn-, Pb-contaminated, mixed contaminated, and control samples. Threshold method was applied to extract the significant variables from LIBS spectra. Then, LRC was used to classify the different types of T. granosa. Other classification models and feature selection methods were used for comparison. LRC was the best model, achieving an accuracy of 90.67%. Results indicated that LIBS combined with LRC is effective and feasible for T. granosa heavy metal detection.     

神经性贝类毒素免疫亲和柱的制备与应用

通过纯化PbTX-2单克隆抗体,以琼脂糖凝胶(sepharose 4B)为载体制备神经性贝类毒素(NSP)免疫亲和柱,并建立NSP的免疫亲和柱净化-液相色谱-串联质谱分析方法。样品以含80%甲醇水溶液提取,PBS缓冲液稀释,经过NSP免疫亲和柱净化和富集后进行LCMS/MS测试分析。同时对上样液、淋洗液、洗脱液等亲和柱参数进行优化。结果表明:NSP免疫亲和柱对贝类样品有很好的净化作用,降低了贝类基质中脂肪、蛋白质、色素、分泌黏液等杂质干扰,提高了样品回收率。在40~800μg/kg范围内样品加标平均回收率为82.95%~112.95%,相对标准偏差为1.27%~2.94%。该NSP免疫亲和柱净化效果强、灵敏度高且受基质干扰小,适用于贝类中神经性贝类毒素的分析测定。     

舟山市海产贝类和食源性腹泻病例中肠道腹泻病毒监测

目的了解肠道腹泻病毒在舟山市海产贝类的分布和食源性腹泻病例中的感染特征以及两者相关联系,为预防和控制食源性疾病提供有效的对策和措施。方法采用荧光定量聚合酶链式反应(PCR)方法对466份舟山市海产贝类和422份食源性腹泻标本中诺如病毒(NoV)GⅠ型/GⅡ型、轮状病毒(RV)、札如病毒(SPV)、星状病毒(AsV)和肠道腺病毒(AdV)5种主要肠道腹泻病毒核酸进行检测。结果 466份海产贝类样品病毒总阳性率为18.03%(84/466),其中NoV GⅡ型为4.08%(19/466),SPV为9.67%(45/466),AsV为2.79%(13/466),AdV为1.50%(7/466);春季和冬季病毒阳性率较高,农贸市场的牡蛎病毒阳性率最高(24.75%,25/101),养殖毛蚶病毒阳性率最低(10.12%,17/168),不同的病毒阳性率以及不同来源和不同季节贝类的病毒阳性率差异均有统计学意义(P0.05)。422例腹泻患者粪便标本总阳性率为15.64%(66/422),NoV为4.74%(20/422,以GⅡ为主),RV为4.74%(20/422),SPV为3.55%(15/422),NoV GⅡ型、RV和SPV的阳性率相对较高,春季和冬季病毒阳性率较高,不同病毒阳性率和不同季节病毒阳性率差异有统计学意义(P0.05)。结论舟山市海产贝类中肠道腹泻病毒的污染状况较为严重,与食源性腹泻病例中的病毒感染特征存在相关联系,其流行季节和优势病毒类型相同。     

秦皇岛海域食用贝类重金属污染情况分析

为了解秦皇岛海域食用贝类中重金属的污染水平及其存在的健康风险。采用电感耦合等离子体发射质谱法(ICP-MS)和原子荧光光谱法测定了该地区15种常见贝类肌肉和内脏中As(无机砷和总砷)、Pb(铅)、Cr(铬)、Cd(镉)、Hg(甲基汞)的含量,并采用污染指数法、系统聚类法评价了各重金属的污染特征和分类。结果表明:海虹(贻贝,Mytilus edulis)、栉孔贝(Chlamys farreri)、生蚝(Ostrea gigas thunberg)和毛蚶子(Scapharca subcrenata)等的重金属综合污染(MPI)较高。根据NY 5073-2006 无公害食品水产品中有毒有害物质限量标准进行重金属评价。秦皇岛海域食用贝类六种重金属含量最高为Cd,均值为1.129 mg/kg,超出NY 5073-2006 无公害食品水产品中有毒有害物质限量标准,污染较严重,总As和无机As均值分别为0.161、0.021 mg/kg。Cr和Hg均值分别为0.107和0.013 mg/kg,均未受污染,Pb均值为0.188 mg/kg,污染较轻。但部分贝类Cd污染比较严重。其中内脏污染比肌肉部分严重,尤其是大小海螺(Busycon canaliculatu)。因此,需进一步研究秦皇岛海域可食用贝类中Pb和Cd的污染来源并进行有效防治。     

乐清湾北部及嵊泗区域养殖贝类肌肉中镉含量调查及健康风险评估

目的 进行乐清湾北部及嵊泗区域养殖贝类肌肉中镉含量调查并探讨通过摄食途径可能导致的健康风险。方法 测定2016～2018年间采自乐清湾北部缢蛏、青蛤及嵊泗区域紫贻贝等三种养殖贝类肌肉中镉含量水平,与国家限量标准进行比对,并运用单因子污染指数(Pi)评估样品中镉污染程度,并以目标危害系数(THQ)和目标致癌风险(TR)为考量指标,评估通过摄食途径可能导致的健康风险。结果 三种肌肉样品中镉检出率为100%,但超标率为0。含量平均值总体比较：紫贻贝﹥缢蛏﹥青蛤。结论 本研究中所有样品Cd 单项污染指数的Pi 值均远低于1.0 ,绝大部分为正常背景水平,少量呈轻度污染水平。健康风险评估结果显示,所有样品THQ 值均小于安全基准值1.0,数据表明暴露人群没有明显的非致癌健康风险。TR的评价结果均在美国环保署推荐的可接受风险值(10_^-6～10_^-4)范围内,但也有部分样品超过了国际辐射防护委员会(ICRP)推荐的最大可接受风险值(5.0×10_^-5),需引起关注。     

2015~2016年河北省贝类水产品中诺如病毒污染状况研究

目的 对2015~2016年河北省6个设区市的市售牡蛎、海虹、血蚶样品中的诺如病毒进行连续一年的检测, 了解河北省贝类水产品中诺如病毒污染状况。方法 分离贝类水产品消化腺, 用蛋白酶K消化后进行前处理, 提取RNA, 用一步法实时荧光逆转录聚合酶链式反应法检测GⅠ型和GⅡ型诺如病毒。对河北省贝类水产品的诺如病毒污染状况进行分析。结果 共采集691份贝类样品进行检测, 其中仅检出GⅠ型阳性样品16份, 阳性率2.32%; 仅检出GⅡ型阳性样品22份, 阳性率3.18%; 同时检出GⅠ和GⅡ型的阳性样品4份, 阳性率0.58%; 总阳性率6.08%。2016年6~7月阳性率最高。沿海地区样品阳性率为内陆地区的2.35倍, 有统计学差异(χ2=8.58, P<0.01)。结论 河北省市售贝类水产品中部分存在诺如病毒污染, 需要加强对贝类水产品的诺如病毒污染检测和控制, 特别是沿海地区, 降低诺如病毒引起的食源性腹泻的疾病负担。     

Matrix effects on a cell-based assay used for the detection of paralytic shellfish toxins in bivalve shellfish samples

Detecting marine biotoxins such as paralytic shellfish toxins (PSTs) is essential to ensuring the safety of seafood. The mouse bioassay is the internationally accepted method for monitoring PSTs, but technical and ethical issues have led to a search for new detection methods. The mouse neuroblastoma cell-based assay (Neuro-2a CBA) using ouabain and veratridine (O/V) has proven useful for the detection of PSTs. However, CBAs are sensitive to shellfish-associated matrix interferences. As the extraction method highly influences matrix interferences, this study compared three extraction protocols: Association of Official Analytical Chemists (AOAC) 2005.06, AOAC 2011.02 and an alternative liquid–liquid method. These methods were used to assess the matrix effect of extracts from four commercially important bivalve species (Chilean mussel, Magellan mussel, clam and Pacific oyster) in Neuro-2a CBA. Extracts from all three protocols caused a toxic effect in Neuro-2a cells (without O/V) when tested at a concentration of 25 mg of tissue-equivalent (TE) ml^?1. The greatest toxicity was obtained through the AOAC 2011.02 protocol, especially for the Chilean mussel and Pacific oyster extracts. Similar toxicity levels (less than 15%) were observed in all extracts at 3.1 mg TE ml^?1. When assessed in Neuro-2a CBA, AOAC 2005.06 extracts presented the lowest matrix interferences, while the highest interferences were observed for AOAC 2011.02 in Magellan mussel and clam extracts. Finally, the AOAC 2005.06 and alternative protocols were compared using Chilean mussel samples fortified with 40 and 80 µg STX per 100 g meat. The AOAC 2005.06 method demonstrated better results. In conclusion, the AOAC 2005.06 extracts exhibited the fewest interferences in the Neuro-2a CBA. Therefore, this extraction method should be considered for the implementation of Neuro-2a CBA as a high-throughput screening methodology for PST detection.     

酶联免疫法快速检测贝类中腹泻性贝类毒素

应用美国ABRAXIS腹泻性贝类毒素(DSP)试剂盒对广西钦州和防城港海域近江牡蛎各20个样品、北海海域文蛤20个样品进行了ELISA法DSP检测.全部检测过程在2h内完成,检测限为0.1 ppb,灵敏度为1.7 ppb.ELISA法检测DSP探作方便、成本低,检测限和灵敏度均满足要求.ELISA法有望作为理想的DSP...     

全自动氨基酸分析仪法测定4种贝类肌肉中谷胱甘肽和游离氨基酸含量

建立一种测定贝类肌肉中谷胱甘肽和游离氨基酸的全自动氨基酸分析仪法,以4种常见贝类为研究对象,采用0.02 mol/L盐酸溶液作为提取剂,体积分数5%磺基水杨酸溶液为蛋白沉淀剂,同时优化了缓冲溶液的组成及洗脱程序,并对该方法的线性范围、检出限、精密度以及回收率进行测定。结果表明:贝类肌肉中的谷胱甘肽和游离氨基酸有着较好的分离效果,线性关系良好,相关系数为0.999 1~0.999 9,检出限(RSN=3)为0.07~0.27μmol/L,加标平均回收率为86.40%~102.42%,日内和日间相对标准偏差分别为0.31%~0.73%和1.14%~2.60%。4种贝类肌肉组织中的游离氨基酸含量比较丰富,总游离氨基酸含量分别为1 396.39、1 016.04、911.15、287.01 mg/100 g,文蛤青蛤缢蛏牡蛎,主要的游离氨基酸为牛磺酸(Tau)、谷氨酸(Glu)、丙氨酸(Ala)、精氨酸(Arg);谷胱甘肽含量分别为103.20、82.53、61.77、33.37 mg/100 g,青蛤文蛤缢蛏牡蛎。该方法适用于贝类肌肉中谷胱甘肽和游离氨基酸含量的测定与分析。     

Effect of steam cooking on distribution of paralytic shellfish toxins in different tissue compartments of scallops Patinopecten yessoensis

This study investigated the effect of steam cooking on distribution of paralytic shellfish poisoning toxins (PSP-toxins) in scallops Patinopecten yessoensis. Toxins analysis by high-performance liquid chromatography showed that most of the PSP-toxins (>70%) were accumulated in viscera and adductor muscle of the raw scallops. Steam cooking induced significant loss (p < 0.05) of PSP-toxins from viscera (16%), adductor muscle (24%), gill and mantle (11%) while 32% of the toxins were retained inside viscera and adductor muscle. Overall, 51% of PSP-toxins leaked out from scallop tissues during steam cooking. However, there was no significant loss (p > 0.05) of PSP-toxins from gonad. Consumption of viscera was the most significant risk factor for causing PSP, while gonad and scallop soup were the second most significant. A rapid PSP test further verified detectable levels of PSP-toxins in all samples. However, possible interfering substance(s) in adductor muscles and gonads might potentially affect the results from this test.