链状亚历山大藻暴露下紫贻贝体内麻痹性贝毒蓄积转化规律

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。     

Modelling current speed and carrying capacity in long-line blue mussel (Mytilus edulis) farms

The development of the mussel (Mytilus edulis) farming industry in Norway is based on suspended long‐line culture, and large areas of the coast are potentially suitable for farming. Norwegian fjords and coastal waters are regarded as oligotrophic environments in comparison with sites where most studies on mussel feeding on natural seston have been carried out. High mussel culture densities in oligotrophic water may cause seston depletion, resulting in low growth or tissue wasting due to reduced feeding and negative net energy balance. In this paper we present a carrying capacity model based on rate conditional processes, balanced against flushing and with emphasis on flow reduction as a function of farm design. The model is based on assumptions that friction forces are a function of geometric shape of the channel made up by the suspended mussel ropes as vertical boundaries and it quantifies carrying capacity according to information of farm length, space between long lines, seston concentration and background current speed and the relative importance of these factors. Estimates of how stocking density in mussel farming can be optimized in relation to the food supply (i.e. carrying capacity) are crucial to production management decisions, and the model may provide predictors for decisions regarding new site selection or expansion of existing operations.     

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.     

Optimization of blue mussel (Mytilus edulis) seed culture using recirculation aquaculture systems

By introducing recirculation aquaculture systems (RAS) in the nursery phase of the blue mussel (Mytilus edulis) (17–18 mm), we aimed at a similar growth and survival and a similar water quality compared to the commonly used flow‐through systems (FTS). To calculate water flow and size of the biofilter, a series of experiments were done to determine clearance rate (9.26 mL min^?1), pseudo faeces threshold (60 000 cells Pavlova lutheri mL^?1), nitrogen production (0.00065 mg TAN h^?1 ind^?1 and 1.6 × 10^?5 mg NO₂–N h^?1 ind^?1) and oxygen consumption (0.03 ± 0.01 mg O₂ h^?1 ind^?1). RAS showed no significant differences in water quality (0.06 mg TAN L^?1; 7.7 mg O₂ L^?1) and growth performance of mussel seed specific growth rate (SGR = 5% day^?1) after the experimental period of 4 weeks compared with FTS. The low water refreshment, 10% per day, as well as the constant chlorophyll concentrations (9.76 ± 1.06 μg L^?1), suggests the potential of RAS as culture system for mussel seed.     

Growth variations within a farm of mussel (Mytilus galloprovincialis) held near fish cages: importance for the implementation of integrated aquaculture

Fish farming releases extensive amounts of particulate organic waste that can be exploited by bivalves in integrated culture. We tested if mussels Mytilus galloprovincialis cultured at two depths (1 and 6 m) in a raft, moored 170 m from a fish farm had greater growth than bivalves held 550 m from the fish cages. Mussel growth was monitored monthly, covering the second phase of the culture, from thinning‐out to harvest (March to November 2011). We also studied if fish solid and dissolved nutrients increased the organic content of the seston and chlorophyll‐a levels near the fish cages through weekly samples. Results showed no differences in seston, chlorophyll and physiochemical characteristics of the water among rafts. Maximum growth and Condition Index (CI) occurred during spring–summer (April–August), when mussels had access to greater food quality and quantity. Mussels cultivated close to the cages showed similar shell length, weight and CI compared with mussels distant from the fish farm. Average shell length, meat dry weight and CI at harvest were 76.31 mm, 2.51 g and 23%. Bivalves cultured distant from the fish cages displayed 26% higher biomass than the other raft at the end of the experiment. Differences in biomass were explained by the significantly higher recruitment of mussel seed observed at the raft distant from the fish cages from June to November. The lack of a significant enhancement in growth of the bivalves cultured next to finfish is discussed.     

DNA fragmentation in blue mussel (Mytilus edulis) sperm: aquaculture and fisheries implications

The objective of this study was to assess sperm DNA longevity in blue mussels (Mytilus edulis) using a dynamic assessment of sperm DNA fragmentation (SDF) after sperm activation. Mature blue mussels (n = 57) in Vigo (Galicia, Spain) were obtained, specifically rope farmed blue mussels (n = 38) and wild blue mussels (n = 19). After the sperm collection, a subsample was assessed for SDF (0 h), while the rest of the sample was incubated for 6, 9, 12, 24 and 48 h at 15°C, assessing each time point using the Sperm‐Halomax kit (Halotech DNA, Madrid, Spain). The Kaplan–Meier estimator, log‐rank (Mantel–Cox) test and Mann–Whitney U‐test were used for statistical analyses (spss v. 16.0), α = 0.05. The rate of SDF (r‐SDF) between rope farmed and wild blue mussels over 0–6 h incubation was not significantly different (P = 0.278), but was for 6–24 h (P = 0.004). Differences in r‐SDF were observed when comparing the means between the two groups (P < 0.0001). Individual differences in r‐SDF existed among the rope farmed (P < 0.0001) and wild blue mussels (P < 0.0001). Wild blue mussels presented a higher DNA longevity than the farmed blue mussels. Selection of blue mussel males with a low level of sperm DNA damage and greater sperm DNA longevity may result in better fertilization and seed production.     

Culture of the blue mussel Mytilus edulis (Linnaeus, 1758) fed both phytoplankton and zooplankton: a microcosm experiment

With a 10‐week microcosm experiment, we demonstrated that the mussel Mytilus edulis could feed and grow upon zooplankton, phytoplankton and mixture of them. The group supplied with the mixture showed the highest shell growth rate, egestion rate and largest size of faecal pellets. Individuals feeding on seawater (the control group) had the lowest growth rate, egestion rate and smallest size of faecal pellets. Egestion rates and faecal sizes of all the groups decreased with experimental time. Therefore, the mussel M. edulis could derive energy from many kinds of diet particles. Most of these particles within the water column may play an important role in bivalve nutrition, feeding and aquaculture. A mixed diet of phytoplankton and zooplankton yielded better growth performance and metabolism than diets of each fed separately.     

Heritability of body size in the Chilean blue mussel (Mytilus chilensis Hupé 1854): effects of environment and ageing

Selection, either natural or artificial, induces an inter‐generational response only when the trait exhibits additive genetic variation. An index of additive genetic variation is narrow‐sense heritability (h²): the ratio between additive genetic variance and phenotypic variance. Here, we present narrow‐sense heritabilities for mass and size in the Chilean blue mussel Mytilus chilensis at different ages and in three different populations. We found that h² changed with age and that maternal‐non‐additive variance was high, generating relatively low estimates of h² in most cases. Our results, together with previous studies, suggest that genotype‐by‐environment interaction does not appear to play a significant role. However, we found that non‐additive and common environmental variance are important determinants of phenotypic morphological variation in body size in M. chilensis. Further studies are needed to establish the precise contributions of these sources of variation, to assess the real potential for response to selection.     

A molecular tool for parentage analysis in the Mediterranean mussel (Mytilus galloprovincialis)

Estimation of selection response and genealogical tracing in family mixtures require an appropriate tool for parentage analysis. In this study, we tested 19 marker loci for parentage analysis allocation in Mediterranean mussel (Mytilus galloprovincialis). To this aim, we reared families in tanks isolated from wild mussel seed, analysed them using the 19 marker loci and characterized their performances based on Mendelian rules. Probabilities of exclusion of a false parent were estimated for different groups of loci and contrasted to the real paternity assignment. Based on this, we chose nine microsatellites with the highest exclusion probabilities and a real paternity assignment of 99.6%. Next, we analysed 600 individuals reared as in the usual production process, where contamination from wild seed is likely. We obtained a real assignment of 94.7% and were able to identify individuals from the wild as the most likely hypothesis to explain the observed incompatibilities with candidate parents. This information was used to evaluate parental contribution in offspring obtained from gamete mixtures of several parents, which bestowed results of interest for future breeding programs of Mediterranean mussel.     

Heritability estimates of larval and spat shell height in the Chilean blue mussel (Mytilus chilensis Hupe 1854) produced under controlled laboratory conditions

The heritability of larval size is estimated in Mytilus chilensis, based on a nested design, involving 95 full‐sib and 19 half‐sib families (19 males each mated to 5 separated females) grown under controlled laboratory conditions. The half‐sib heritability estimates at 10, 25 and 40 days for larval and spat shell height ranged between 0.38±0.33 and 0.84±0.45, suggesting that selection for shell size would be effective.     

Shell-shape variation along the latitudinal range of the Chilean blue mussel Mytilus chilensis (Hupe 1854)

Shell‐shape variation in the Chilean blue mussel Mytilus chilensis was examined in eight populations covering the totality of its distribution range, which represents over 1800 km of latitudinal gradient in the southeastern Pacific. The shell outline shapes were assessed using elliptic Fourier analysis followed by canonical variates analysis. Fourier coefficients showed a highly significant morphological variation between the populations studied. Canonical variates analysis showed a visual association of the First canonical with convexity of ventral edge and umbo shape, while the second axis was associated with shell elongation. Previously described genetic patterns were marginally congruent with our morphometric pattern, though geographic distance had a stronger effect on morphology. Shape change (elongation) was correlated with latitudinal clinal variation. The morphometrical analysis proved to be an important tool for evaluating the components of shell‐shape variation in mussels and to document association patterns with geographical and ecological variables. Such patterns are useful to establish a wide field of work, including sexing, stock recognition, traceability studies and productive management in general.     

Evaluation of growth performance and intestinal barrier function in Arctic Charr (Salvelinus alpinus) fed yeast (Saccharomyces cerevisiae), fungi (Rhizopus oryzae) and blue mussel (Mytilus edulis)

Arctic charr (Salvelinus alpinus) were fed for 99 days on experimental diets with 40% of fish meal replaced, on a crude protein basis, with intact yeast (Saccharomyces cerevisiae) (ISC), extracted yeast (ESC), Rhizopus oryzae fungus (RHO) or de‐shelled blue mussels (Mytilus edulis) (MYE). The fish were evaluated for growth performance, nutrient digestibility and fish intestinal function. Growth performance, retention of crude protein and sum of amino acids were not affected in fish fed diets ISC or MYE compared with those fed the reference (REF) diet. However, fish fed diet ISC displayed decreased digestibility of crude protein and indispensable amino acids and decreased intestinal barrier function compared with fish fed the REF diet. Fish fed diet ESC exhibited decreased growth performance and protein retention, but had comparable digestibility to fish fed the REF diet. Fish fed diets MYE and RHO showed similar performance in terms of growth, nutrient digestibility and intestinal barrier function. Overall, the results indicated that blue mussel and intact S. cerevisiae yeast are promising protein sources for Arctic charr.     

Predation by the polyclad flatworm Imogine mediterranea on the cultivated mussel Mytilus galloprovincialis in Bizerta Lagoon (northern Tunisia)

This study was carried out to investigate a paradigm of predation under natural conditions in Bizerta lagoon. The free‐living marine polyclad flatworm Imogine mediterranea was found to feed on the commercial mussel Mytilus galloprovincialis. Such predation could represent a real threat to mytiliculture at this site. Our field experiments have shown that the mortality of the commercial mussel as a result of active predation by this polyclad is approximately 52% when one polyclad and one prey were placed together in a same experimental container. We found a statistically significant relationship between the size of the polyclad and the extent to which the mussels were disabled or completely eaten. Moreover, in our field experiments, the times required for predation‐caused decreases of populations of the bivalves, that is, LT₁₀, LT₂₅ and LT₅₀, decreased significantly with increases in the size of the polyclad. The time, LT₅₀, required for polyclads to kill half of the bivalve population in field experiments was approximately 4.5 days. A simple disabling (an attack by the polyclad on the adductor muscle of the mussel) was shown to be sufficient to cause irreversible changes leading to mortality. Furthermore, disabling and mortality were found to be linearly correlated. The importance of mucous secretions, pharyngeal type and body musculature in supporting predation by polyclads is discussed. We suggest that measures to control the polyclad pest could incorporate the control of this flatworm's embryonic and postembryonic developmental stages.     

Effects of environmental factors and food availability in Northern Aegean sea on the cultivation of Mediterranean Mussels (Mytilus galloprovincialis)

This study used two different collectors made of polypropylene (PP) rope and polyethylene (PE) fishing net to determine Mediterranean mussel's settlement, growth, condition index (CI), meat yield (MY) and shell thickness index (STI) during the period spent between June 2017 and November 2018. With this regard, chlorophyll a, water temperature, salinity, pH and particulate matter were defined through water samples that were collected on monthly basis triplicate. The mean chlorophyll a level was recorded as 0.32 ± 0.31 μg/L, while water temperature was 19.73 ± 5.14°C, salinity was 35 ± 2.070‰, pH was 8.12 ± 0.04, and TPM was 14.91 ± 10.48 mg/L. As a result of the cultivation period of 8 months spent following the first intense grip, the length of the mussels is measured as 31.79 ± 6.20 mm. Based on the Pearson correlation analysis results, the most important environmental parameter affecting Mediterranean mussels growth in length on PP rope collectors is the temperature (p < .05). The STI and CI were determined to be related with environmental temperature parameters, while no determinations could be obtained concerning the correlation of MY with environmental parameters. Assessment of Mediterranean mussels’ growth rate in PP rope collectors proved significant differences (p < .05) between March and November 2018. As no Mediterranean mussels are observed on PE fishing nets during the samplings performed throughout the study period, it is determined that PE fishing nets are not suitable collector types for Mediterranean mussel cultivation.     

厚壳贻贝人工繁殖技术的研究

了解厚壳贻贝 (Mytilus coruscus) 的生活环境对其生长的影响,对贻贝养殖业的高质量发展极为重要。以枸杞岛潮间带野生和浮筏养殖厚壳贻贝为研究对象,构建以壳长为基准的13个体框特征指标 (包括壳宽、壳高等),比较两种厚壳贻贝的主要形态特征,并结合线粒体16S rRNA 基因的序列信息,分析其遗传关系与形态特征的关联性。结果表明,野生和养殖贻贝有10个体框特征指标存在显著性差异 (P<0.05),野生的贝壳整体更宽厚、质量更大。应用逐步判别法筛选出的壳宽 (L₁)、壳高 (L₂)、壳顶至铰合部上端的距离 (L₁₀)、壳顶至足丝孔的距离 (L₁₂) 4个变量对野生和养殖厚壳贻贝的综合判别准确率达到94.9%。16S rRNA基因测序结果显示,野生厚壳贻贝的核苷酸多样性 (π) 为0.089,单倍型多样性 (H_d) 为0.894;养殖厚壳贻贝的π为0.087,H_d为0.682。野生与养殖厚壳贻贝的遗传多样性水平均较高,但系统发育树和单倍型网络图证明二者不存在显著的遗传分化,推测其形态差异可能主要受栖息环境 (波浪暴露强度、营养条件) 影响。

     

Growth and mortality of Mediterranean mussel Mytilus galloprovincialis Lam., 1819, in relation to size on longline in Mersin Bay, Izmir (Turkey – Aegean Sea)

The growth and mortality rate of different size of mussels, Mytilus galloprovincialis, were compared in Mersin bay, Izmir. Mussel sampling was performed on a monthly basis over 1 year, together with hyrobiological parameters. Water temperature, salinity, chlorophyll a, total particulate matter (TPM) and particulate organic carbon (POC) were determined. Average chlorophyll a, TPM and POC values were 3.88±1.62 μg L⁻¹, 13.12±3.68 mg L⁻¹ and 252±121.89 μg L⁻¹ respectively. Initial mean lengths of mussels for each size group were 10 mm (9.91±0.25 mm), 20 mm (20.14±0.29 mm), 30 mm (30.66±0.21 mm), 40 mm (40.14±0.21 mm), 50 mm (50.62±0.21 mm), 60 mm (59.77±0.21 mm) and 70 mm (69.47±0.62 mm). Maximum growth in length occurred during the spring–summer months. The annual length increments of mussel size groups were 46.22, 41.42, 30.3, 28.03, 20.63, 16.34 and 12.03 mm from small to large size groups respectively. Small mussels grew faster than large mussels and reached commercial size at the same time. Mortality was <5% for all groups at the end of the experiment.     

Novel decaplex PCR assay for simultaneous detection of scallop species with species‐specific primers targeting highly variable 5′ end of the 16S rRNA gene

Scallops (family Pectinidae) comprise species of high commercial value, supporting both commercial fisheries and mariculture activities. Accurate and reliable molecular methods for the species level identification are of outstanding utility for taxonomic and food authentication surveys. The mitochondrial 16S rRNA gene has been used to design species‐specific primers for identification of different bivalve species. However, the low interspecific variability at the 3′ end of this gene has limited its utility and only few scallop species have been assessed. In this study, we used the high variable 5′ end of the 16S gene to develop a novel decaplex PCR assay that enabled a fast and accurate identification of eight commercially important scallop species in a single PCR reaction. A total of 285 individuals including fresh and manufactured samples from eight different processed presentations from 11 different scallop species were collected representing diverse locations around the world. Our assay accurately identified all the analysed samples at the species level. Furthermore, to enhance the utility of our assay, the PCR product amplified by the family specific primer set that was utilized as positive control was also used for the identification of unknown (non‐target) scallop species by DNA sequencing analysis. In its present form, our multiplex PCR method can be of great utility for different types of studies involving scallop species and for research institutes and governmental agencies that regulate seafood authentication around the world.     

The genetic structure of Mytilus chilensis (Hupé 1854) populations along the Chilean coast based on RAPDs analysis

The Chilean blue mussel Mytilus chilensis is an important commercial species. However, little has been published on the population genetics of this species, despite the need to implement management and conservation policies. Randomly amplified polymorphic DNA‐polymerase chain reaction analysis was used to estimate genetic variation within and between eight natural populations along the whole range of its Chilean natural distribution (ca. 1900 km from Arauco (VIII Region) to Punta Arenas (XII Region)). The values of Nei's unbiased genetic distance, D (0.030–0.107), among populations were small, despite the large geographic separation. A mantel test using 50 000 randomizations showed evidence for a significant correlation (r=0.74, P<0.05) between genetic and geographic (coastal) distance. Punta Arenas population was the most genetically differentiated from the others, although the scale of differentiation was not large (D=0.076–0.107). The levels of gene flow (Nm=1.55) found in this study prevent differentiation among populations by genetic drift. This is the result of the long‐lived planktotrophic larvae of M. chilensis, which provides this species with considerable dispersal ability throughout its range, which is favoured by the ocean currents along the Chilean coast. A restricted larval dispersal towards the north due to the Cape Horn Current derived from the West Wind Drift could be the cause of the higher genetic differentiation of Punta Arenas population from the northern populations. For management purposes of the M. chilensis fishery, the results provide no evidence for discrete stocks, with the possible exception of the Punta Arenas population. The present study provides the baseline data in order to continue further characterization of these mussel populations, considering the great increase in aquaculture of this species.