间接免疫荧光法检测栉孔扇贝急性病毒性坏死症病毒

用纯化栉孔扇贝(Chlamys farrei)急性病毒性坏死症病毒(AVNV)免疫兔子,以兔抗血清为一抗,荧光标记的羊抗兔抗体为二抗,采用冰冻切片技术,建立了栉孔扇贝AVNV的间接免疫荧光检测方法。用该方法分析栉孔扇贝和海湾扇贝(Argopecten irradians)体内AVNV感染强度,并对感染率进行统计。结果表明,栉孔扇贝的肾脏、肝胰腺中,AVNV阳性信号最强,呈现中度到重度感染,其AVNV感染率100％。鳃丝、性腺及闭壳肌中未检测到阳性信号。在海湾扇贝的肝胰腺及肾中AVNV的感染率也较高。提示AVNV感染扇贝的靶器官主要是肾脏、肝胰腺。对与栉孔扇贝同一海区养殖的海湾扇贝在栉孔扇贝发病期间存活率较高的原因进行了讨论。     

First report of signs of infection by Coccomyxa-like algae in wild blue mussels,Mytilus spp., in the Gulf of Maine (USA,Maine)

In August 2019, visual inspection of intertidal zones of the Gulf of Maine (ME, USA) revealed young and adult wild blue mussels, Mytilus spp., in Alley Bay (Jonesport area) with the distinctive L-shaped shell deformity (LSSD) and green spots (GS) in the mantle and adductor muscle. LSSD is a characteristic sign of current or previous mussel infection by photosynthetic unicellular alga from the group Coccomyxa, while GS are algal colonies. Based on these findings, this study represents the first report of infection signs by pathogenic Coccomyxa-like algae in mussels from the coastal waters of the Northeastern United States, providing a base for future large scale monitoring of the alga in the region.     

Bioaccumulation and interrelationship of manganese and zinc in horse mussel Modiolus modiolus fromEastern Canada

ABSTRACT:   Bioaccumulation and distribution of Mn and Zn in the total softtissues, digestive glands, residuals and adductor muscles of thehorse mussel Modiolus modiolus from three sites, includingindustrialized and non-industrialized locations in Eastern Canada,were investigated. Extremely high digestive gland metal concentrationswere found in individual mussels, as high as 1819 µg/gMn wet weight and 1964 µg/g Zn wet weight,with mean values from 358 to 404 µg/gMn and from 399 to 614 µg/g Zn for thecollection sites. High Mn to Zn interrelationships were observed inall types of tissues and at all sites. Between different tissues,Zn was interrelated by linear relationships, and Mn was best describedby power curve relationships for all tissue types. In the totalsoft tissue, Mn and Zn interrelations were fitted to power regressioncurves with different slopes between the three study sites. Thisindicated that horse mussel was exposed to different metal levels inthe environments and could be useful for monitoring these metals.The uptake of both metals at extremely high concentrations, thelack of regulation and the occurrence of interactions all suggest thatMn and Zn may play a biological role in horse mussels. Zn and Mninteractions, surprisingly, were not disrupted at the very highconcentrations of either metal, which proves that the mechanism ofmetal interactions does not involve a detoxification role.     

紫贻贝4种同工酶的组织特异性表达研究

采用聚丙烯酰胺垂直电泳法对紫贻贝Mytilu sedulis的鳃、消化腺、外套膜、足、闭壳肌等5种组织的超氧化物歧化酶（SOD）、三磷酸腺苷酶（ATPase）、苹果酸脱氢酶（MDH）、苹果酸酶（ME）进行了检测分析。结果表明,SOD在紫贻贝的5种组织中均有表达,共有4条酶带,但迁移率最大的酶带在足和外套膜中未出现,而在另外3种组织中呈弱表达;ATPase在除消化腺外的组织中都表现为2条带;MDH在5种组织中都表达出1条酶带,其中足和闭壳肌的MDH染色最深;ME存在1～2条酶带,其中迁移率大的条带在5种组织中都有表达,而迁移率小的条带只存在于外套膜、闭壳肌和足中。从检测结果来看,上述4种酶在紫贻贝体内的表达有一定的组织特异性．     

Influence of microalga lipid composition on the sexual maturation of Mytilus galloprovincialis: a hatchery study

The effect of microalga lipid composition on the reproduction of the mussel Mytilus galloprovincialis has been assessed to determine the best feeding strategy for producing large quantities of mussel seed. Three diets based on two microalgae Isochrysis galbana (clon T‐iso) and Chaetoceros gracilis were tested. Besides, hatchery groups were compared with mussels from natural populations. Lipid content and fatty acid profile of digestive gland and mantle of both sexes, eggs and microalgae were analysed and related to sexual maturation. Hatchery groups, specifically the one fed on T‐iso, showed better results in reproduction success, and these differences were reflected on tissue and egg lipid composition. Microalga fatty acid profile influenced tissues and sexes, and higher levels of 18:1n‐9, 22:6n‐3 (DHA) and 18:2n‐6 were detected in groups fed on T‐iso while higher level of 20:5n‐3 (EPA) and 16:1n‐7 in groups fed on C. gracilis. Evidence of synthesis capacity of EPA from 18:4n‐3 and DHA from EPA is detected comparing their levels and the mobilization between tissues. Egg fatty acid profile was influenced by the female diet, and differences among groups were detected and confirmed by PCAs.     

Study of the Toxic Contaminants Cadmium and Lead in Sardines (Sardina pilchardus) in the Bay of Algiers (Algeria)

The study of the discharge of metallic pollutants into the marine environment through biological material such as sardines makes it possible to evaluate the quality of the biological resources and of the marine environment. The sardine (Sardina pilchardus) is the most popular species for fishing and the most consumed species by the Algiers population. Two toxic metals that can affect human health are studied in this fish. A year of sampling with fishermen was conducted. Sampling was done in the spring, in the autumn, and in April between April 2014 and April 2015. Sampled sardines were grouped by size classes. The gills and muscle were recovered by size classes. The results were explored using a principal components analysis and compared by a Mann-Whitney distribution test. An atomic absorption spectrophotometer was used in the laboratory for analysis. The most important results show that lead levels were higher than those of cadmium in the gills and muscle. For sardines, lead contamination appears to have both gill and food origins. Mann-Whitney comparison tests confirmed this homogeneous distribution of lead between the gills and muscle. The correlation between cadmium in the gills and height or weight is more substantial than that with muscle. The origin of cadmium in sardines is branchial. The food origin appears secondary. From our point of view, the effect of season, i.e., winter (December) and spring (April), favors an increase in metal levels in parallel to the production reserves planned for spawning. The effect of season was observed by use of a biplot. A significant reduction in cadmium and lead content was observed in September. The high temperature, an environmental factor, induced stress that reduced the metal concentrations and the decontamination induced by the issuance of genital products in early summer.     

虾夷扇贝组织中微量元素的分布特性

研究了虾夷扇贝闭壳肌、外套膜、内脏团、瓣鳃和性腺中Cu、Fe、Mn、Zn、Pb和Cd等元素的分布特性。试验结果表明,内脏团中的Cu显著(P<0.05)高于其他各组织,其他各组织间差异不显著(P>0.05);闭壳肌中Zn的含量显著(P<0.05)高于内脏团、外套膜、瓣鳃和性腺中的含量;Fe含量内脏团和瓣鳃中显著(P<0.05)高于性腺、闭壳肌和外套膜;各组织中Mn的分布特性为瓣鳃>闭壳肌>性腺>内脏团>外套膜;Pb的分布特性为内脏团>闭壳肌>性腺>外套膜>瓣鳃;内脏团中的Cd占全贝总量的67%,显著高于其他组织。因此,约占全贝质量10%的内脏团蓄积了较高含量的Cu、Fe、Pb和Cd,尤其是Cu和Cd(分别约占全贝蓄积总量的71%和67%),食用时去掉内脏团,可保证虾夷扇贝的食用安全和较高的食用及营养价值。     

Lipid composition of the mantle and digestive gland of <Emphasis Type="Italic">Octopus vulgaris</Emphasis> juveniles (Cuvier, 1797) exposed to prolonged starvation

Lipid composition of the mantle and digestive gland of Octopus vulgaris that were not fed for 27 days were determined. Every 3 days, three octopuses were killed and samples of the mantle and the digestive gland (DG) were taken, in order to determine total lipids as well as lipid classes and fatty acids. Composition in total lipids (TL) for the mantle was similar until day 21, then decreased and remained similar until the end of the experiment. Composition in total lipids for the DG decreased significantly after 3 days, then remained similar until day 21, and then decreased until the end of the experiment. As for the lipid classes, in the DG the main components were triglycerides and sterol esters. Sterol esters suffered strong reductions after 10 days of starvation, while triglycerides remained similar until day 21 and then decreased until the end of the experiment. Cholesterol decreased gradually throughout the experimental period. For polar lipids, phosphatidylcholine and phosphatidylethanolamine increased during the first 3 days and then decreased throughout the experiment. In the mantle, the only neutral classes that decrease were triacylglycerols and sterol esters, while no polar lipid classes decreased in this organ. It was noticeable the decrease in almost all fatty acids in the DG after 3 days of starvation, while in the mantle there were no differences in fatty acid concentrations during the experiment.     

Digestive dynamics during chyme formation of Octopus maya (Mollusca,Cephalopoda)

In the present study, digestive dynamics, measured through the changes in the in situ pH along the digestive tract, was evaluated along with the glycogen concentration in the digestive gland (DG) and the proteolytic enzyme activity of the gastric juice (GJ) (both acid and alkaline proteases) to obtain useful information that will allow for the understanding of the DG's function and the role that it plays as an extracellular digestive enzyme source and for energy storage, providing new information on the digestive physiology of Octopus maya. The results showed that pH along the digestive tract changed according to the postprandial time following the food transit. At the beginning, the pH on crop (St) was 5, changing to 6 when the food arrived. Similar changes were observed on the caecum (Ce) and DG. Glycogen from DG is used as a source of energy during digestion and recovered 8 h after feeding. Maximum activities of digestive gland GJ enzymes were observed 6 h after feeding, indicating that chyme enzymes are still active when they arrive at the DG's lumen. The presence of food in the digestive tract modifies the pH, which, in combination with GJ, favours the activity of the released enzymes of O. maya.     

Changes in lipid composition of different tissues of common octopus (Octopus vulgaris) during short‐term starvation

In this work, we study the variations in concentration (mg 100 g^?1 dry weight) and total content (mg individual^?1) of different lipid classes in muscle and the digestive gland of Octopus vulgaris during an 8‐day starvation period. In all the samples analysed, polar lipids (PL) predominated in muscle (267.3–337.2 mg 100 g^?1) compared with neutral lipids (66.9–104.7 mg 100 g^?1). A significant positive tendency was observed in muscle for the content and concentration of monoglycerides as a consequence of starvation (P < 0.05). Furthermore, a higher cholesterol (CHO) content was detected in this tissue after the fourth day of starvation compared with the fed animals (P < 0.05). Neutral lipids predominated in the digestive gland (12 958–14 151 mg 100 g^?1) compared with PL (3157–6517 mg 100 g^?1), with triglycerides, free fatty acids and monoglycerides being the major lipid classes. The concentration of PL and monoglycerides showed a positive trend with starvation, while the triacylglycerol showed a negative tendency (P < 0.05). The results suggest that triglycerides and CHO are transported simultaneously from the digestive gland to the muscular tissues during starvation and the preferential use of PL and CHO during growth phases. It was estimated that lipids contributed 26% of the energy costs of the animals during starvation, mainly in the form of triglycerides from the digestive gland.     

Seasonal Changes in Quality and Fatty Acid Composition of Black Mussel (Mytilus galloprovincialis)

The study was designed to assess the seasonal variations in the quality and lipid profile of mussel meat (Mytilus galloprovincialis) harvested along the Bulgarian coast of the Black Sea. The trial period lasted from June to October, and the sampling was carried out in the area of Varna Bay. Technological quality of the mussels was determined by measuring the water holding capacity (WHC) and cooking and roasting losses. Proximate composition, total aerobic plate count, and fatty acid analysis were carried out. Technological parameters and nutritional quality of the mussel meat were strongly affected by the season. During the summer, the mussels had higher mass, meat content, and meat yield (p < 0.001) compared to the early autumn, which corresponds to the lowest protein content during this season. Lipid profile showed seasonal variations in both individual and total fatty acid amounts (p < 0.001). Saturated fatty acids (SFA) had the highest content in summer, while monounsaturated fatty acids (MUFA) decreased gradually until the autumn. The content of polyunsaturated fatty acids (PUFA) reached its peak in October, contributing to the most favorable values of the nutritional indices of lipids in the early autumn.     

The interaction of infectious salmon anaemia virus (ISAV) with the blue mussel,Mytilus edulis

Integrated multi‐trophic aquaculture (IMTA) is an alternative approach to mono‐culture aquaculture that reduces environmental impacts of commercial aquaculture systems by combining the cultivation of fed species with extractive species. Shellfish play a critical role in IMTA systems by filter‐feeding particulate‐bound organic nutrients. They may also increase or decrease disease risk on farms by serving as reservoirs or barriers for important finfish pathogens such as infectious salmon anaemia virus (ISAV). This study aimed to optimize culture and molecular assays in shellfish tissues and to determine the fate of ISAV in mussels, Mytilus edulis. To determine detection limits, qRT‐PCR and culture assays in both CHSE‐ and ASK cells were optimized in ISAV‐inoculated mussel tissue homogenates. Both qRT‐PCR and culture assays performed in ASK cells had comparable detection limits of 10^2.8 TCID₅₀ mL^?1. The ISAV RNA genome was consistently detected in digestive gland tissue of ISAV‐exposed mussels. Viable ISAV was not detected in mussel tissues by culture analysis in CHSE‐ and ASK cells. The fact that qRT‐PCR analysis resulted in positive cycle threshold (CT) values that corresponded to the detectable range of ISAV in ASK culture assays suggests that little to no viable ISAV particles are present in the mussel tissues.     

Relationships among environment, spawning and settlement of Queen scallop in the Ría de Arosa (Galicia, NW Spain)

This study examines the relationship of the reproductive and reserve storage cycles and spat settlement of the Queen scallop (Aequipecten opercularis) to environmental factors. The possibility of farming this species from spat settled on collectors is considered. The gonad condition index shows maximum values between mid-May and early August. Another peak occurs in mid-November. In spring and summer, the energy needed for gametogenesis is derived from food, although it is possible that direct transfer may occur from the digestive gland to the gonad. The glycogen and lipid content of the reserve organs reaches maximum values between late summer and early autumn. The energy requirements related to gametogenesis, which begins in autumn, involve the consumption of accumulated reserves with the resulting reduction in the condition indices. There is a positive correlation between temperature and the condition indices of the digestive gland, the adductor muscle and their lipid and glycogen content, respectively. Two settlement periods were recorded, one in winter, which could not be associated with the spawning of the experimental animals, and another in summer, which coincided with the spawning that took place in spring. The reduced gonad indices recorded at the end of the summer and in winter did not lead to settlement.     

扇贝内脏团中耐镉菌株的分离及其吸附镉机理

为探讨扇贝内脏团中微生态系统与扇贝高能力富集镉(Cd)的关系,从自然环境中采集的栉孔扇贝内脏团中分离、纯化了2株耐Cd细菌(编号为菌株A和B),运用16S rDNA基因序列进行分子鉴定,通过金属吸附实验研究了耐Cd菌株对Cd的吸附能力及其对Cd的吸附特性,并利用扫描电镜、透射电镜相结合的方法研究了Cd胁迫条件下耐镉菌株细胞形态与结构变化,探讨了其对Cd的吸附机理。结果显示,2株菌株(A和B)在固体培养基上能耐受Cd的浓度分别为100和80 mg/L。经16S rDNA测序鉴定菌株A与Nitratireductor sp.亲源关系最近,菌株B与Ruegeria sp.亲源关系最近。2株菌株对Cd的吸附率远高于铜(Cu)、锰(Mn)、锌(Zn)和铅(Pb)等重金属。在50 mg/L Cd浓度的液体培养基中,菌株A、B对Cd富集量分别为48.57和42.14 mg/g,富集系数分别为971.4和842.8。电镜观察结果显示,经过Cd处理后,2菌株数量均有所减少,细胞中出现空泡,菌株A细胞外沉淀增多,菌株B表面变得粗糙且出现凹陷,表明胞外沉积作用可能是耐Cd菌株对Cd富集作用的重要途径。     

海产品与淡水产品中重金属的差异性及其风险评价

为了解我国东部沿海地区海产品与淡水产品中重金属污染现状及摄入风险,实验分析了东部沿海地区市售的12种海产品和8种淡水产品,采用微波消解-电感耦合等离子体质谱法（ICP-MS）测定铅、镉、汞、砷、铬共5种常见重金属,根据单因子污染指数（P_i）和金属污染指数（MPI）评价其污染程度,比较不同水产品重金属含量差异及污染状况,并通过靶标危害系数（THQs）评价其健康风险。检测结果显示,海产品与淡水产品中,铅含量范围为ND～1.100 mg/kg,铅超标率为1.4%,各品种铅均值含量无显著差异。镉含量范围为ND～1.600 mg/kg,镉超标率为2.8%,长牡蛎、海湾扇贝镉均值含量高于其他品种。总汞含量范围为ND～1.900 mg/kg,蓝鳍金枪鱼总汞均值含量高于其他品种,其他品种总汞均值含量无显著差异。总砷含量范围为0.004～4.100 mg/kg,淡水鱼（乌鳢除外）总砷均值含量低于其他品种。铬含量范围为0.010～32.000 mg/kg,铬超标率为2.3%,鳙铬均值含量高于其他品种。单因子污染指数显示：不同品种的水产品都有一定的污染。重金属污染指数显示：污染指数呈现...     

The effect of diets containing different levels of active charcoal on growth performance,body composition,haematological parameters and possibility of heavy metals detoxification in big sturgeon (Huso huso)

In order to determine the chelating effect of activated charcoal, a total of 150 beluga (270 g) were fed for 60 days with a diet containing 0 (control), 5, 10, 15 and 20 g activated charcoal per kg. Afterwards, the fish were subjected to challenging with 1 mg/L of each of heavy metals including lead, cadmium and copper during seven days. As results, the highest percentage of body weight gain, specific growth rate, condition factor, protein efficiency ratio and the lowest feed conversion ratio before and after challenging with heavy metals were obtained in 15 g treatment with significant differences comparing to other treatment (p < .05). In all treatments, considerable differences were observed between two experimental stages of all parameters except condition factor and feed conversion ratio (p < .05). No mortalities were observed during the experiment. Before challenging with heavy metals, 15 g treatment showed the lowest activities of AST and ALT enzymes, and uric acid, protein, albumin and cholesterol amounts. The minimum amounts of creatinine and glucose were recorded in the control and 20 g treatments respectively. The highest levels of these parameters, except for creatinine and uric acid, were measured in the control group. After challenging with heavy metals, the lowest levels of AST and ALT enzyme activities, uric acid, creatinine, and glucose, and the highest amounts of protein, albumin and cholesterol belonged to 15 g, whereas the results in the control group were almost opposite. In all treatments, significant differences were observed between two experimental stages of all parameters except AST, uric acid, glucose and cholesterol (p < .05). In the pre‐challenge stage, the maximum amount of carcass protein was observed in 15 g while the differences in lipid content were not significant (p > .05). Moreover, the highest contents of moisture and ash were measured in the control group and 20 g respectively. After the challenge, the highest values of protein and lipid were quantified in 15 g, while the maximum values of ash and moisture were observed in 20 g and control group respectively. Also, significant differences were observed just in ash and moisture amounts in both stages (p < .05). Accumulations of cadmium, copper and lead were maximal in the control group in both stages, and their amount were decreased in sample with 15 g of activated charcoal until they reached to their lowest value in sample with 20 g of activated charcoal. The only different parameter between two treatments was copper amount. Also, significant differences were observed in all treatments in both stages (p < .05). Although activated charcoal at 20 g/kg diet yielded the best results in metal chelation, it is recommended to add 15 g/kg of activated charcoal to the diet of beluga because of other parameters.     

Cadmium detoxification effect of Chinese parsley Coriandrum sativum in liver and kidney of rainbow trout Oncorhynchus mykiss

Although fish feed supplemented with Chinese parsley (CP), Coriandrum sativum, does not affect the health of rainbow trout Oncorhynchus mykiss, it has the remarkable effect of reducing the uptake of Cd (12–17% compared with controls without CP). Here, we measured the change in the amount of metallothionein (MT) in the liver and kidney using gel permeation chromatography-high-performance liquid chromatography (TSK GEL SW 3000 column). The increase in MT content over time in the Cd-fed groups corresponded to the change in Cd accumulation. Gel permeation was performed with soluble fractions from the fish liver and kidney, and changes in the Cd, Cu, and Zn concentrations over time for each fraction, and the relationship between the form of Cd that accumulated in tissues and its toxicity were measured. A nontoxic MT fraction of 7,000 Da and a high-molecular-weight fraction of approximately 60,000 Da were isolated from the soluble fractions. The induction of nontoxic MT–Cd was higher in the CP-supplemented groups than in the control groups. The synergetic actions of several compounds in CP may contribute to bind MT, thereby preventing accumulation of harmful heavy metals and essential metals in fish.     

A Novel Metallothionein Gene from Mussel,Hyriopsis cumingii: Identification and Expression under Lanthanum Exposure

Metallothionein (MT) is a superfamily of cysteine‐rich, metal‐binding, low‐molecular‐weight proteins that participate in the storage, transportation, and metabolism of the trace metal elements. In this study, a novel MT gene from Hyriopsis cumingii was cloned (designated HcMT1); it shared 59.2% identity with previously published MT sequence of H. cumingii and 56.1–97.3% identity with other MT genes from mollusks. The expression level of HcMT1 in various tissues was determined by real‐time polymerase chain reaction. Relatively higher expression levels were observed in digestive gonad, mantle, and gill, and the expression level in hemocytes was significantly upregulated by lanthanum (La³⁺) exposure. The nucleotide sequence of HcMT1 amplified from genomic DNA is identical to the cDNA template group under La³⁺ exposure, which indicated that HcMT1 is an intronless gene. These results indicate that HcMT1 may play an important role in La³⁺ metabolism.     

Upstream recolonization by freshwater mussels (Unionoida: Hyriidae) following installation of a fishway

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Marteiliosis in mussels: a rare disease?

Among 1280 cultured and wild adult Mediterranean mussels, Mytilus galloprovincialis, collected over a 1 ‐ year surveillance period from the Slovene Adriatic Sea, 0.3% were histologically positive for the presence of Marteilia spp. The infection was concentrated in winter. Employing the molecular methods of PCR, cloning, DNA restriction and sequencing, only Marteilia refringens type M was detected in all the infected mussels. Although all life‐cycle stages of M. refringens severely infected digestive glands, only sporadic disruption of epithelial cells of digestive tubules and focal destruction of digestive tubules were observed in the infected mussels. This was the first detection of M. refringens in M. galloprovincialis from the Slovene Adriatic Sea with the lowest prevalence reported to date. In addition, our results highlight the need for sequencing to complement the established PCR‐RFLP analysis for correct parasite typing.