太平洋牡蛎中大肠杆菌净化的实验研究

静水中人工感染太平洋牡蛎,使其体内埃希氏大肠杆菌(Escherichia coli)浓度达到105 MPN/100g,然后放入紫外线循环系统中对其进行净化,研究贝水质量比(1∶3,1∶6,1∶10,1∶50)、水交换频率(1,3,5次/h)、温度(10,15,20,25℃)及净化层数(上层,中层,下层)等不同环境因子对牡蛎体内大肠杆菌净化的影响。在前6h大肠杆菌净化迅速,之后速度减慢,至36h时能净化2～3个对数值。不同环境因子的净化实验结果表明,当贝水比为1∶6,水交换律为3次/h,温度为15或25℃,净化层数为上层时净化效果最佳。     

繁茂膜海绵清除大肠杆菌的过程

通过分析海绵清除大肠杆菌的过程,研究海绵净化细菌的机理。作者利用荧光显微镜和激光共聚焦显微镜观测等手段,监测和分析了绿色荧光大肠杆菌(Escherichia coli)在繁茂膜海绵(Hymeniacidon perlevis)体内、体外水环境中数量变化过程。在1 L含有3×107个/m L绿色荧光大肠杆菌的海水中放入鲜重(1.02±0.11)g的繁茂膜海绵24块,处理7 h,海水中的荧光大肠杆菌数量逐渐降低;而海绵体内荧光大肠杆菌数量在2 h时内逐渐增多,之后的2 h趋于稳定,4 h以后开始逐渐减少。水体中大肠杆菌不仅进入海绵体内,而且进入海绵细胞内。含有荧光大肠杆菌的海绵块转入无菌海水中后,海绵体内及细胞中大肠杆菌逐渐消失,而且大肠杆菌没有被释放到环境海水中。分析表明,繁茂膜海绵能够以摄食的方式净化水环境中的大肠杆菌。     

钦州湾海水和香港牡蛎体内脂溶性贝类毒素污染特征

近年中国沿海普遍受到脂溶性贝类毒素(lipophilic shellfish toxins,LSTs)的污染。钦州湾是国内外闻名的牡蛎养殖基地,目前仍不了解该海域海水和主要养殖品种香港牡蛎(Crassostrea hongkongensis)沾染LSTs毒素情况与食用安全性。本文于2015年10月至2016年9月,在钦州湾香港牡蛎养殖区海水交换口处,通过固相吸附毒素跟踪技术(solid phase adsorption toxin tracking,SPATT)吸附收集海水中的脂溶性贝类毒素,同时每月采集香港牡蛎样品,应用液相色谱串联质谱法分析海水和香港牡蛎体内的LSTs毒素含量与组成,并对香港牡蛎食用安全性进行评估。结果表明:(1)海水共检出GYM、OA和PTX2三种组分,平均浓度分别为0.44、14.20和1.67μg/(kg resin·30d)。香港牡蛎共检出DTX2、GYM、SPX1、Homo-YTX四种组分,其中Homo-YTX为定性结果, DTX2、GYM、SPX1三者平均浓度分别为0.95、18.86和0.95μg/kg。(2) GYM毒素均值在海水中和香港牡蛎体内含量变化趋势呈正相关,Pearson相关系数为0.70;OAs毒素在海水中和香港牡蛎体内变化趋势类似, Pearson相关系数为0.35。(3)毒素存在一定季节变化。海水OA和PTX2夏季高,其他季节低,均值最高值分别发生在7月和6月;香港牡蛎体内GYM冬季和夏季较高,DTX2春季和夏季较高;但海水中的GYM和香港牡蛎体内的SPX1全年变化均不明显。总体而言,钦州湾海水和香港牡蛎已受到多个LSTs毒素组分污染,海水LSTs毒素含量较国内其他海域低,参考欧盟LSTs毒素标准,本海域的香港牡蛎脂溶性贝类毒素仍未超标,可放心食用。     

厦门西海域海水和养殖动物污染物初步调查

1999年2月和4月对厦门西海域海水和两种养殖动物的重金属（Cu、Zn、Cd、Pb)、石油类和大肠杆菌数量进行了调查。结果表明：西海域海水的重金属（Cu、Zn、Cd、Pb)、石油类含量基本符合国家一类海水水质标准，养殖动物鲈鱼和僧帽牡蛎也基本符合国家食品卫生标准。海水中的大肠杆菌数量超过了国家海水卫生标准，显示该海域有一定程度的有机质污染，其养殖产品不宜生食。马銮湾内海水和养殖动物的污染物含量较海外高。养殖鲈鱼的肝脏对重金属和石油类的浓集系数远较肌肉为高，而僧帽牡蛎对污染物的浓集能力又明显比鲈鱼强。     

海水工厂化养殖废水循环利用的初步探讨

以海水工厂化养殖排出的废水养殖双齿围沙蚕(Perinereis aibuhitensis)、菲律宾蛤仔(Ruditapes phi-lippinarum)、长牡蛎(Crassostreagigas)和细基江蓠(Gracilariatenuistipitata).结果表明:双齿围沙蚕耐污染能力最强;细基江蓠和双齿围沙蚕具有较强的净化水质能力.养殖废水经7 d处理后,NH4+-N、NoO3--N、NO2--N、PO43--P、COD和底质有机物分别下降至处理前的11.20%、23.69%、27.50%、14.6%、3.2%和0.32%,且沙蚕和江蓠平均日生长率分别为2.30 mg/g·d和80 mg/g·d.该方法不但能净化水质,还能实现养殖废水的循环利用.     

膜集成技术在海水养殖中的应用与探索

采用超滤、紫外辐射、臭氧氧化三种形式的多效滤除、混合协同作用，展示并模拟出了应用膜集成技术预防养殖用海水水质恶化的最佳技术路线，通过可行性试验表明，超滤膜过滤与臭氧氧化作用均有可靠的杀菌效果，组合工艺不但保持了养殖所需的盐氛围，且可以增加水中的溶解氧，净化水质，从而提高海水的重复利用率，节省能源。     

一株海洋芽孢杆菌B09的筛选及其发酵条件优化研究

从海水养殖环境中筛选出一株具有较强抗菌活性的芽孢杆菌B09,并对其进行抗菌活性研究和发酵条件优化。结果显示B09对金黄色葡萄球菌、大肠杆菌、鳗弧菌等致病菌有较强抑制作用,其最佳发酵条件如下:KB培养基(蛋白胨20g,甘油10mL,K2HPO41.5g,MgSO4.7H2O1.5g,水1000mL),发酵温度32℃,发酵起始pH为7,发酵时间48h,250mL的三角瓶装量100mL,接种量1%。     

细基江蓠繁枝变种与益生菌净化养殖废水的研究

通过室内实验和室外排水沟实验研究了细基江蓠繁枝变种Gracilarla tenuistipitatavar.liui与益生菌(光合细菌Rhodopseudomonas capsulata和芽孢杆菌Bacillus lichenifirmis24h内对集约化对虾养殖废水的协同净化作用。室内实验显示益生菌和细基江蓠繁枝变种对灭菌后的养殖废水有净化作用,净化效果表现为:混合组(江蓠 益生菌)>江蓠组>益生菌组>对照组;室外排水沟实验中江蓠组与混合组都显示对养殖废水的净化有明显作用,但两组之间各项指标(除TN外)差异不显著(p>0.05),而益生菌组与对照组的各项指标之间的差异也不显著(p>0.05)。这并不能否定江蓠与益生菌之间的协同作用,而可能与排水沟水体的异养细菌数高(检出数为104—105CFU.ml-1)有关。     

不同长牡蛎群体对海水酸化的生理响应差异分析

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。     

四倍体与二倍体太平洋牡蛎离体精子的存活能力比较

在不同海水温度和不同精子密度条件下对四倍体(4n)与二倍体(2n)太平洋牡蛎(Crassostrea gigasThunberg)离体精子的存活能力进行了比较.结果表明,四倍体与二倍体太平洋牡蛎精子的存活时间均随水温的升高和密度的降低而缩短,四倍体牡蛎精子的存活能力低于二倍体.四倍体牡蛎精子在海水温度10℃可存活8 h,在28℃仅存活2 h;二倍体牡蛎精子在10℃可存活12 h,在28℃仅存活4 h.四倍体牡蛎精子在较高密度(6.4×10<'7>个/Ml)下可存活8 h,较低密度(2.0×106个/Ml)下低于1 h;在与四倍体牡蛎精子相同的高、低密度下,二倍体存活时间分别超过10 h和1 h.     

Depuration dynamics of oysters (Crassostrea gigas) artificially contaminated by Salmonella enterica serovar Typhimurium

The State of Santa Catarina produces the greatest quantity of edible mollusks in Brazil. To guarantee sanitary qualify, mollusk cultures should be monitored for contamination by pathogenic microorganisms. A self-purification or "depuration" system that eliminates Salmonella enterica serovar Typhimurium contamination from oysters has been developed and evaluated. The depuration process occurred within a closed system, in which 1000 L of water was recirculated for 24 h. The water was sterilized with ultraviolet (UV) light, chlorine, or both together. Oysters (Crassostrea gigas) artificially contaminated with S. typhimurium were harvested every 6 h. Samples of oyster tissue were excised and both the presence and numbers of bacteria were determined. Combined UV light and chlorine treatments resulted in total elimination of bacteria within 12 h. Polymerase chain reaction detected bacteria in water exposed to the three treatments. This pioneering study is the first of its kind in Brazil and represents a major contribution to commercial mollusk culture in this country.     

香港巨牡蛎抗副溶血弧菌感染相关microRNA的筛选

香港巨牡蛎（Magallana hongkongensis,旧称Crassostrea hongkongensis）是我国华南沿海重要的经济贝类,但频发的养殖病害严重制约了香港巨牡蛎养殖业的可持续发展。作为一类重要的调控因子,microRNA（miRNA）能够对其靶基因进行转录后调控从而参与海洋动物各类生命活动过程。本研究利用高通量测序技术建立了副溶血弧菌不同诱导时间下的香港巨牡蛎鳃组织miRNA文库,并通过生物信息学分析方法挖掘与免疫调控有关的miRNA,共获得了453个已知miRNA和222个新miRNA,通过进一步筛选空白对照组/12 h病原注射组、空白对照组/24 h病原注射组、12 h病原注射组/24 h病原注射组特有的miRNA,发现多个在不同比较组间差异表达的或具有高表达水平的miRNA。其中miR 10、miR 23、miR 92、miR 100、miR 125、miR 144、miR 145、miR 146、miR 335、miR 750等已被证实参与了多种海洋动物的免疫调控过程。对差异表达miRNA靶基因的KEGG富集分析结果显示miR 411 5p、miR 4454 5p、novel m0159 3p、novel m0187 3p的靶基因涉及免疫相关通路。分析结果表明上述miRNA可能在香港巨牡蛎免疫调控过程中发挥重要作用,有必要对其开展进一步研究。     

The potential food poisoning hazard of Vibrio parahaemolyticus in New Zealand Pacific oysters

The level of Vibrio parahaemolyticus was generally very low in Pacific oysters sampled from 4 New Zealand oyster farms between November 1982 and May 1983. Of the 149 samples collected, only one was found to contain greater than the 1000 organisms g^‐1regarded as the maximum allowable by international standards (International Commission on Microbiological Specifications for Foods 1982: Micro‐organisms in Foods, Vol. 2. Academic Press, New York). V. parahaemolyticus was detected in 57% of the oyster samples, but 95% of these contained less than 10 organisms g^‐1. Maximum levels appeared to coincide with high water temperatures at the farm sites. Chilling, freezing, and depuration reduced V. parahaemolyticus numbers in oysters after harvest. However, holding at ambient temperatures (19–25°C) resulted in up to a 35‐fold increase in organism numbers with a maximum after one day. The food poisoning hazard from V. parahaemolyticus in New Zealand‐grown Pacific oysters thus appears to be minimal. Recommendations are made to keep harvested shellfish cool and to chill them as soon as practical.     

Feasibility of co-culture of the Australasian sea cucumber (Australostichopus mollis) with the Pacific oyster (Crassostrea gigas) in northern New Zealand

The Australasian sea cucumber (Australostichopus mollis) has attracted commercial attention for aquaculture development, partly due to its potential for co-culture with shellfish and finfish species. However, minimal attention has been given to the possibility of co-culturing this species with oysters. In this study we evaluated the growth of juvenile sea cucumbers (36.7 ± 0.9 g, wet weight) caged underneath Pacific oyster farms in northern New Zealand. Co-culture started at the end of the summer, and after 304 days the juveniles had doubled in size (79.8 ± 3.3 g, wet weight), but their subsequent growth appeared to be constrained by overstocking of the cages and summer water temperatures, reaching a carrying capacity of 720 g m^?2. Overall, the results of this study indicate that the co-culture of juvenile sea cucumbers with Pacific oysters is feasible, if sea cucumber losses are reduced (between 33% and 52% lost in this study) and careful attention is given to stocking rates and the water temperature regimes of oyster farms in order to maintain adequate growth rates.     

养殖密度、水层和养殖地点对马氏珠母贝选育群体生长存活的影响

用通过壳高定向选择培育的第三代马氏珠母贝Pinctada martensii为实验材料，研究不同的养殖密度、养殖水层和地点对生长和存活的影响。实验结果表明在一定试验范围内养殖密度对体重增长有显著影响（p<0.05），高密度组（200只/笼）的生长较低密度组慢，密度与水层对生长无明显的交互作用。水层对生长的影响不显著（p>0.05），但对存活率的影响明显，2m水位的存活率较高，且水层与养殖密度间有交互作用。不同地点的贝的生长无显著性差异（p>0.05）。该研究结果将为珍珠贝优良品种（系）养殖技术的优化和品种推广体系的建立提供指导。     

A non-invasive anaesthetic method for accessing the brood chamber of the Chilean flat oyster (Ostrea chilensis)

A cost effective method to discern brooding from non-brooding specimens of the Chilean flat oyster (Ostrea chilensis Phillipi, 1844) is needed to optimise current hatchery practices for this species. Accordingly, we investigated the efficacy of two treatments, AQUI-S® and MgCl₂, in anaesthetising individuals of this species and then mapped any subsequent mortality of anaesthetised oysters. Oysters immersed for 3 h in solutions of 20, 40, 80 and 100 µL L^?1 of AQUI-S® exhibited no signs of anaesthesia and also recorded low levels of mortality (i.e. 8% or less). Conversely, oysters immersed in 30, 40 and 50 g L^?1 MgCl₂ showed between 96%–100% anaesthesia after 3 h whereas only 70% of oysters immersed in 60 g L^?1 MgCl₂ were anaesthetised over this time period. Time taken for 50% of oyster shells to open (SO₅₀) was compared among MgCl₂ treatments with oysters immersed in 60 g L^?1 taking significantly longer than (103 min) those immersed in 30–50 g L^?1 (SO₅₀ values of 43, 54 and 37 min respectively). Furthermore, treating oysters with MgCl₂ resulted in low levels of mortality (i.e. 13% or less) and was not related to MgCl₂ concentration. Thus, our results show that immersing O. chilensis in MgCl₂ to be an efficient and cost effective method to access the brood chamber of this species. We recommend the use of 30 g L^?1 for routine anaesthesia and recommend that future work investigates the efficacy of cheaper MgCl₂ sources and what effects MgCl₂ anaesthetisation has on developing larvae.     

冷冻牡蛎贮存过程中脂溶性有毒物质的变化研究

牡蛎是我国出口量较大的水产品之一,由于产品本身的特殊性,在生长过程中易产生腹泻性贝毒(DSP).本实验中原料牡蛎DSP生物法检测结果为阴性,冷冻贮存4个月后检测结果即为阳性.本研究以分析这一变化原因为目的,采用LC-MS法对DSP生物检测的阳性样品进行确认,结果表明,生物法DSP检测的结果为假阳性,造成生物法DSP检测中小白鼠死亡的是牡蛎在贮藏过程中产生的脂溶性有毒物质.进一步分析确定该有毒物质为牡蛎脂肪水解物--游离脂肪酸.同时本研究对牡蛎在不同贮存温度和贮存时间下产生的游离脂肪酸含量进行了测定,在-10 ℃贮存温度下,贮存0,2,4,6,8,10个月,游离脂肪酸的变化范围为5.2～788.6 mg/kg;在-10 ℃,-18 ℃,-25 ℃贮存温度下,贮存4个月,游离脂肪酸的变化范围为92.5～389 mg/kg.     

中国牡蛎产业的嬗变——新认知、新品种和新产品

牡蛎是近海生态系统的重要成员,也是世界性海洋主养贝类。我国牡蛎养殖历史悠久,但高质牡蛎产品的长期匮乏已成为新形势下产业转型升级的卡点。在国际海鲜市场,高质牡蛎就意味着品质好、品相优。为实现产业高质量发展,研究了中国经济牡蛎物种组成和地理分布;揭示了温度是不同尺度种性形成的重要环境驱动因子之一;构建了首个贝类全基因组序列精细图谱,发现基因组的高变异性和基因家族的特异性扩张是种性形成的重要遗传基础;对全球27个长牡蛎群体487个个体进行全基因组深度重测序,构建了50M级单核苷酸多态(SNP)资源库并制成190k高密度SNP分型芯片。这些资源基因组学(Resourceomics)研究为高质牡蛎创制奠定了基础。其次进一步查清了牡蛎经济性状的遗传力及表型相关性,锚定糖原含量调控的基因组模块区域,建立品质性状基因模块选育技术,育成"海蛎1号"新品种,糖原含量提高25.37%,比传统育种效率提高65.81%,实现了单一营养物质的定向选育,破解了牡蛎肉质改良的世界性难题。所建立的基因模块育种技术使高质牡蛎遗传创制成为可能。在育成新品种的基础上,还利用牡蛎附着变态阶段的生物学特性及上升流和下降流的物理学原理,创新牡蛎单体种苗制备技术,使种苗单体化率提高3倍;建立设施塑形生态育肥技术,通过壳型重塑并结合养殖水层的增肥调控,优型率达92%,出肉率达20%—23%。单体塑形养殖技术使牡蛎品相也达到国际知名品牌的产品标准。团队建立的高质牡蛎创制技术体系在县域规模进行标准化应用示范,支撑了"乳山牡蛎"成为行业第一品牌,产业经济效益提高2—3倍,实现了牡蛎产业从低质低效到高质高效的嬗变,示范带动了中国牡蛎产业的高质量发展。     

Response of phytoplankton to multispecies mariculture:a case study on the carrying capacity of shellfish in the Sanggou Bay in China

A muhispecies model for shellfish polycuhure in the Sanggou Bay in China used for large-scale long-line cultivation of the Chinese scallop Chlamysfarreri, the Pacific oyster Crassostrea gigas and the kelp Laminaria japonica is presented. The model includes key physical processes which are the transports of matter at the system boundary, and the main biological process that is the primary production and nutrients release from the bottom. By the model, the seasonal fluctuations of phytoplankton biomass and dissolved inorganic nitrogen（DIN） in 1994 are simulated. Furthermore, if the kelp culture scale is kept constant and the Chinese scallop and the Pacific oyster culture scales are adjusted, virtual shellfish farms are funded and responses of phytoplankton to the largescale shellfish culture are simulated. According to these simulated results, the room limitation, and the hypothesis that shellfish will not grow well if the phytoplankton biomass is less than 8.2 mg/m^3 , the expandable multiple of scallop culture k and that of oyster culture y are determined as k = -0.276 5y ＋4.690 5 and 0.133 3k ＋0.006 6y≤0.667 5, where, k （ or y） is equal to 1, the culture scale of scallop （ or oyster） is 8.8 x 109 individuals （or 66 ha, with a density of 59 ind./m^2 ）, and the kelp culture scale is 3 300 ha with a density of 12 ind./m^2.