四种/株亚历山大藻(Alexandrium)毒性的比较研究

通过对塔玛亚历山大藻(ATHK和AT-6藻株)、链状亚历山大藻(ACDH藻株)和微小亚历山大藻(AM-1藻株)的PSP毒素含量和组成以及它们分别对轮虫、黑褐新糠虾和鲈鱼存活影响的比较研究发现:塔玛亚历山大藻AT-6藻株不产生麻痹性贝毒毒素PSP,而塔玛亚历山大藻ATHK、链状亚历山大藻ACDH和微小亚历山大藻AM-1均产生PSP毒素,其总含量分别为19.74、5.395、5.57fmol/cell(2.60、0.36、1.61pgSTXeq/cell)。塔玛亚历山大藻AT-6对轮虫、黑褐新糠虾和鲈鱼这三种生物均没有不利影响;对于轮虫,塔玛亚历山大藻ATHK和链状亚历山大藻ACDH的96hLC50分别为:200和1200cells/ml,而微小亚历山大藻AM-1无不利影响;对于糠虾,塔玛亚历山大藻ATHK、链状亚历山大藻ACDH和微小亚历山大藻AM-1的96hLC50分别为7000、11000、16000cells/ml;对于鲈鱼,这三株藻的96hLC50分别为3700、4000、20000cells/ml。四株亚历山大藻对三种生物的毒性大小与其PSP毒素含量和组成无直接的相关关系,其毒性作用可能来自其他未知毒性物质。塔玛亚历山大藻ATHK不同组分分别对三种生物的毒性比较研究表明:藻细胞重悬液和藻液对三种生物的不利影响最为显著,去藻过滤液和细胞碎片对轮虫和鲈鱼没有明显影响,但糠虾的存活率有所下降;细胞内容物对轮虫和糠虾也无影响,但对鲈鱼的存活有一定影响。以上研究表明:不同亚历山大藻的毒性大小存在差别,其对不同生物的危害机制也不相同,除PSP毒素外,亚历山大藻可能还存在其他的毒性物质。     

海南岛的海涂土壤及其开发利用

根据调查资料和采样分析,作者划分海南岛海涂土壤类型,并就机械组成、养分含量、盐渍化程度、粘土矿物组成等方面阐述海涂土壤特征。根据海涂土壤的性质及其生态环境特征进行开发利用,宜以海水养殖和晒盐为主,其中海水养殖可分为8个区。     

基于UPLC-Q-Exactive MS构建麻痹性贝类毒素的特征指纹溯源技术

麻痹性贝类毒素污染是世界共同关注和重点管控的食品安全问题, 其溯源一直是产业监管的难点和技术重点。基于指纹溯源技术理论, 采用室内模拟3种产毒藻暴露紫贻贝后风险形成过程, 开发基于超高效液相色谱-四极杆-静电场轨道阱高分辨质谱(UPLC-Q-Exactive MS)鉴定麻痹性贝类毒素区域特征的指纹溯源技术。结果表明, 采用乙腈/甲醇/丙酮(体积比为1︰1︰1)混合溶液作为提取剂, C8色谱柱联合正离子模式, 质荷比(m/z) 100~600和Amide色谱柱联合正离子模式, m/z 600~1 500作为色谱质谱检测条件下, 提取的化合物数量较多且稳定性良好, 覆盖了40.4%指纹信息; 紫贻贝与产毒藻中的共检出11种毒素成分, 且具有高度相关性。N-磺酰胺甲酰基类毒素(如C类)和膝沟藻毒素(GTX类)在暴露期发生了显著的代谢转化; 将摄食3种产毒藻后紫贻贝的差异化合物与特征毒素组分进行主成分分析及正交偏最小二乘法判别分析, 共筛选出13种复合指纹物质。构建6种特征物质的Fisher判别模型, 交叉验证准确率为88.9%, 可实现产毒藻种溯源。该研究建立了基于超高效液相色谱-四极杆-静电场轨道阱高分辨质谱UPLC-Q-Exactive MS紫贻贝特征指纹溯源技术, 初步实现了从污染麻痹性贝类毒素的贝类到肇事藻种的逆向溯源,可应用于贻贝麻痹性贝类毒素风险溯源研究。     

Clean and safe supply of fish and shellfish to clear the HACCP regulation by use of clean and cold deep ocean water in Rausu, Hokkaido, Japan

For the supply of fish and shellfish to consumers in fresh condition, clean handling after catch from the sea is essential. According to HACCP （Hazard Analysis and Critical Control Points）, it is important to meet such requirement by keeping fish and shellfish under a certain low temperature and clean conditions after catching. The deep ocean water （DOW） characterized by low temperature and cleanliness has been chosen for fish and shellfish handlings, particularly for salmon, cod, and sea urchin in Town ＇Rausu＇ in Hokkaido, Japan. DOW below 2.9℃ of an amount of nearly 5 000 m^3 is planned to be pumped up every day from a depth of about 350 m, and temporarily stored in a large simulated tank on land. DOW is then supplied to fish boats through hydrants distributed throughout the harbor and used for keeping salmon in clean and cold conditions. Ice made from DOW is also used for lowering temperature if necessary. DOW and ice made from DOW are also used during the transportation of fish and shellfish. The entire system is scheduled to be completed by the summer of 2005.     

UHPLC-LTQ-Orbitrap-MS非定向筛查贝类中氮杂螺环酸毒素及其代谢产物

为了分析复杂贝类和微藻中的氮杂螺环酸毒素（AZAs）及其代谢产物,建立一种基于超高效液相色谱-线性离子阱/静电场轨道阱组合式高分辨质谱（UHPLC-LTQ-Orbitrap-MS）联用技术的非定向筛查分析方法。采用Full MS/dd MS²、PRM和Target-SIM/dd MS²三种质谱鉴定模式,整合多种数据采集挖掘策略,通过研究多种AZAs在不同基质样品中的质谱裂解规律及特征碎片离子丰度,实现AZAs及其衍生物的快速筛查检测和精准鉴别。结果表明：PRM模式能有效排除AZAs同分异构体的干扰,获得更高的专属性,适用于目标代谢物筛查;应用Full MS/dd MS²和PRM结合的方式,根据AZAs裂解途径及多种AZAs代谢产物裂解规律,在贝类中共鉴别出20种AZAs系列化合物,其中包括初步推测了3种新型AZAs代谢产物的结构。应用本方法还发现AZA9、AZA10、AZA19等代谢物均随代谢过程持续升高,是AZA2在贝类代谢过程中的末端产物。该方法能够为复杂基质中的AZAs系列毒素及其衍生常规检测与精准鉴别提供参考,可应用于解析AZAs毒素在贝类体内的代谢转化机制研究。     

基于3种生物指数的三沙湾养殖活动底栖环境效应研究

为考察我国东海区最为典型养殖海湾三沙湾养殖活动的底栖环境效应,本研究根据2016年8月航次41个样品采集站位的数据,分析香农-威纳多样性指数（Shannon-Wiener index,H'）、海洋生物指数AMBI（AZTI's Marine Biotic Index）和多变量海洋生物指数M-AMBI（Multivariate-AMBI）在4种典型水域（网箱、海藻、鲍鱼养殖和无养殖水域）间的差异及其与沉积物环境因子（δ15N、δ13C、C/N比、总有机碳、总氮、总磷、硫化物、含水率和粉粒-黏粒比例）的相关性。结果表明,航次共采集74种大型底栖生物,H'介于0~3.51;物种多属环境敏感类型（AMBI方法中EGⅠ和EGⅡ生态组）,AMBI指数介于0~4.13,M-AMBI指数介于0.30~0.77。3种生物指数的分布水平表明此海域总体底栖生态质量良好,仅在局部水域存在显著扰动。Kruskal-Wallis检验显示仅H'在不同类型水域之间存在显著差异（P<0.05）,后续配对检验（Pairwise comparision）显示大黄鱼养殖水域H'显著高于海藻养殖水域（P<0.01）。基于斯皮尔曼秩相关系数（Spearman correlation coefficient）的相关性分析表明仅H'和M-AMBI指数分别与总磷存在显著相关性（P<0.05）。聚类分析（Cluster analysis）结果显示不同类型站位在聚类图中随机分布,聚类结果未体现出采样水域的差异性。综上,基于3种生物指数的分析结果显示三沙湾内网箱等典型养殖活动未对区域底栖生境质量造成显著影响。     

传染性皮下及造血组织坏死病毒(IHHNV)在双壳贝类中的感染情况研究

采用国际兽医局推荐的传染性皮下及造血组织坏死病毒(infectious hypodermal and hematopoietic necrosis virus,IHHNV)检测方法,首次开展IHHNV在双壳贝类(中国蛤蜊、泥蚶、花蛤、缢蛏、文蛤和白蛤)和螺类软体动物(螺蛳、中华圆田螺)中的感染情况。此外,本研究以泥蚶为研究对象,建立了IHHNV在双壳贝类中的实验室感染模型,研究了人工感染泥蚶后IHHNV在其体内的增殖和致病情况。结果显示:在采集的不同种类贝类样品中均检测到IHHNV,其中在泥蚶中的阳性率最高(50%),在文蛤中的阳性率最低(15%),表明IHHNV在双壳贝类中的分布较为广泛,是IHHNV的重要携带物种。在采集的螺类软体动物中均未检测到IHHNV。系统发育分析表明,来自不同贝类中的IHHNV构成了进化树中不同的分支,其中中国蛤蜊、花蛤、白蛤源的IHHNV属于Ⅱ型感染株,而泥蚶、缢蛏、文蛤源的IHHNV单独成簇,形成了一个全新的分支。实验室感染结果显示,IHHNV对泥蚶没有明显的致病性,但可在其体内增殖,在感染后第5天其体内的病毒载量达到最大(1.8×104copies/g),随着感染时间的增加,其体内病毒含量呈现递减趋势,但在感染后第30天体内仍可检测到病毒的存在。本研究结果对对虾养殖尤其是虾贝混养模式中预防和控制IHHNV的传播具有重要意义。     

Comparative study on in vitro transformation of paralytic shellfish poisoning (PSP) toxins in different shellfish tissues

Dissected tissues of three shellfish species,the Chinese scallop,Chlamys farreri,Manila clam,Ruditapes philippinarum,and Razor shell,Solen strictu were evaluated for in vitro transformation of paralytic shellfish poisoning(PSP) toxins. Tissue homogenates were incubated with extraction from toxic algae Alexandrium minutum to determine toxin conversion. The effects of heating and addition of a natural reductant(glutathione) on toxin conversion were also assessed. The toxin profile was investigated through high performance liquid chromatography with fluorescence detection(HPLC-FLD) . The evident variations in the toxin content were observed only in Chinese scallop viscera homogenates. The concentration of GTX4 was reduced by 45%(approximately 0.8 μmol/dm 3 ) and 25%(approximately 1 μmol/dm 3 ) for GTX1,while GTX2 and GTX3 increased by six times(approximately 1 μmol/dm 3 ) and 3 times(approximately 0.3 μmol/dm 3 ) respectively. Simultaneously,the total toxicity decreased by 38% during the 48 h incubation period,the final toxicity was 20.4 nmol STXeq/g. Furthermore,heated Chinese scallop viscera homogenates samples were compared with non-heated samples. The concentration of the GTX4 and GTX1 was clearly 28%(approximately 0.53 μmol/dm 3 ) and 17%(approximately 0.69 μmol/dm 3 ) higher in heated samples,GTX2 and GTX3 were four times(0.66 μmol/dm 3 ) and two times(0.187 μmol/dm 3 ) lower respectively. GSH(+) Chinese scallop viscera homogenates samples were compared with GSH(-) samples,the concentration in the GTX4 and GTX1 was 9%(approximately 0.12 μmol/dm 3 ) and 11%(approximately 0.36 μmol/dm 3 ) lower respectively,GTX2 and GTX3 was 17%(approximately 0.14 μmol/dm 3 ) and 19%(approximately 0.006 μmol/dm 3 ) higher respectively. In contrast,there was a little change in the concentration of PSP toxins of Manila clam and Razor shell tissue homogenates. These observations on three shellfish tissues confirmed that there were species-specific differences in PSP toxins transformation. PSP toxins transformation was more pronounced in viscera tissue than in muscle tissue. PSP toxins was possibly interfered by some carbamoylase enzyme,and the activity in Chinese scallop viscera tissue is more remarkable than in the other two species.     

Fishery resources in arid zone mangroves in gulf of Kachchh, Gujarat, northwest coast of India

The finfish and shellfish resources were assessed quantitatively and qualitatively in regard to their abundance in creek waters at three sites within a period of two years, from January 1999 to December 2000, in the western mangrove areas of Kachchh. The catch rate varied from 0.69 to 6.99 kg h⁻¹. It was low during monsoon (July to October), which could be due to the freshwater-flow-induced salinity reduction in all the sites. Among 38 species recorded, 5 were shellfish and 33 were finfish. The spawning period of fishes was found to be during summer and early monsoon period (May to August). Surface water temperatures varied from 17 °C to 37 °C. Salinity values varied from 34 to 44 and the pH ranged between 7 and 8.9. Variation in dissolved oxygen content was from 3.42 to 5.85 mL L⁻¹. The high fishery densities in these semi arid mangrove creek areas were recorded during monsoon and early winter season.     

An individual-based population dynamic model for estimating biomass yield and nutrient fluxes through an off-shore mussel (Mytilus galloprovincialis) farm

The fluxes of carbon, nitrogen and phosphorus through an off-shore long-line Mytilus galloprovincialis farm during a typical rearing cycle were estimated by combining a simple population dynamic model, based on a new individual model, and a set of field data, concerning the composition of the seston, as well as that of mussel meat and faeces. The individual model, based on an energy budget, was validated against a set of original field data, which were purposely collected from July 2006 to May 2007 in the North-Western Adriatic Sea (Italy) and was further tested using historical data. The model was upscaled to the population level by means of a set of Monte Carlo simulations, which were used for estimating the size structure of the population. The daily fluxes of C, N and P associated with mussel filtration, excretion and faeces and pseudo-faeces production were integrated over the 10-month-long rearing cycle and compared with the total amount of C, N and P removed by harvesting. The results indicate that the individual model compares well with an existing literature model and provides reliable estimations of the growth of mussel specimen over a range of trophic conditions which are typical of the Northern Adriatic Sea coastal area. The results of the budget calculation indicate that, even though the harvest represents a net removal of phosphorus and nitrogen from the ecosystem, the mussel farm increases the retention time of both nutrients in the coastal area, via the deposition of faeces and pseudo-faeces on the sea-bed. In fact, the amount of nitrogen associated with deposition is approximately twice the harvested one and the amount of phosphorus is approximately five times higher. These findings are in qualitative agreement with the results of literature budget and model calculations carried out in a temperate coastal embayment. This agreement suggests that the proper assessment of the overall effect of long-line mussel farming on both the benthic and pelagic ecosystem asks for an integrated modelling approach, which should include the dynamic of early diagenesis processes, as well as of that of nutrients released from the surface sediment.