Effects of environmental oxygen deficiency on embryos and larvae of bay scallop,Argopecten irradians irradians

The experimental results showed that: 1) The embryonic development of bay scallop is inhibited at a dissolved oxygen range of 1.38–3.64×10⁻³ at 23°C, and completely blocked below the lower limit. 2) The tolerance of larvae to anoxia increased with larval sizes and was related to their oxygen debt. 3) The scallop larvae exhibited specific behavioral responses to oxygen deficiency, which finally led to velum disintegration and larval death. The possible relationship between environmental oxygen deficiency and the disease of disintegration of the larval velum is also discussed. In this study, considerable oxygen debt was found in bay scallop larvae, which was greater in small animals. Based on the works of previous authors, a new concept is proposed for the estimation of oxygen debt, namely, the compensatory rate of oxygen debt (CROD). This can be used in intra—or interspecific comparison of oxygen debt. The results can be helpful in the management of water quality and for the prevention of larval diseases encountered in scallop culture. Contribution No. 2167 from the Institute of Oceanology, Chinese Academy of Sciences.     

Comparison of growth and survival of larvae among different shell color stocks of bay scallop Argopecten irradians irradians（Lamarck 1819）

In order to improve production and breed new broods of bay scallop Argopecten irradians irradians, different-colored orange, purple and white lines were established by two mating methods of self-fertilization and mass spawning at a commercial scallop hatchery in spring, 2002. And then larval growth and survival of different lines was compared to test whether there is a relationship between shell color and growth and survival at early developmental stage. Both growth and survival have no significant differences among different experimental larvae of self-fertilization or mass spawning. Results are as following in the order of orange, purple and white shell stock. For the self-fertilization, growth rates of larvae were 6.174, 6.412, and6.599 μm/d, respectively. Survival rates of larvae at Day 3 were 74.41%, 76.86%, and 82.05%; Day 6 were 49.14%, 65.63%, and 52.79%; and Day 9 were 25.06%, 20.80%, and 26.47%, respectively. For the mass spawning, the growth rates were 7.836, 7.941, and 7.878 μm/d, respectively. Survival rates at Day 3 were 93.05%,91.95%, and 92.50%; Day 6 were 79.17%, 78.05%, and 82.50%; and Day 9 were 34.72%, 36.67%, and 38.33%,respectively. The absence of any relationship between shell color and growth and survival at the larval stage may be resulted from their common genetic basis.     

Selective feeding of bay scallop Argopecten irradians on phytoplankton community revealed by HPLC analysis of phytopigments in Bohai Sea,China

Understanding the feeding selectivity on phytoplankton by shellfish is currently a big challenge.In order to investigate the feeding behavior of bay scallop(Argopecten irradians)on phytoplankton,we compared its compositions of phytopigments in digestive glands with those in the surrounding seawater,and conducted five consecutive investigations between July and November 2016 in a bay scallop culture area along coast of Qinghuangdao City,northwest of the Bohai Sea,China.Phytopigments in four-size fractionated phytoplankton of seawater(micro-(20-200 μm);nano(L)-[10-20 μm];nano(S)-[2.7-10 μm],and pico-[2.7 μm])and digestive glands of A.irradians were examined to investigate the selective feeding of A.irradians.Results show that fucoxanthin and peridinin constituted the major part of taxonomically diagnostic carotenoids(TDCs)in the micro-and nano(L)-phytoplankton in seawater.Compared with total phytoplankton biomass of seawater(TPB,sum of the four sizes),a substantial decrease of fucoxanthin proportion to total DCs in digestive glands was observed while that of peridinin,19'-butanoyloxyfucoxanthin,alloxanthin and 19'-hexanoyloxy-fucoxanthin showed an obvious increase when those pigments were mainly confined to micro-sized phytoplankton(20-200 μm).However,zeaxanthin and prasinoxanthin were mainly confined to nano(s)-and pico-phytoplankton,of which the proportions in digestive glands were usually lower in TPB.The contribution of lutein to total DCs in digestive glands(with an average of 7.23%)increased compared with TPB of seawater(with an average of 0.63%)during all five sampling times.     

Comparison of growth and survival of larvae among different shell color stocks of bay scallop Argopecten irradians irradians (Lamarck 1819)

In order to improve production and breed new broods of bay scallopArgopecten irradians irradians, different-colored orange, purple and white lines were established by two mating methods of self-fertilization and mass spawning at a commercial scallop hatchery in spring, 2002. And then larval growth and survival of different lines was compared to test whether there is a relationship between shell color and growth and survival at early developmental stage. Both growth and survival have no significant differences among different experimental larvae of self-fertilization or mass spawning. Results are as following in the order of orange, purple and white shell stock. For the self-fertilization, growth rates of larvae were 6.174, 6.412, and 6.599 μm/d, respectively. Survival rates of larvae at Day 3 were 74.41%, 76.86%, and 82.05%; Day 6 were 49.14%, 65.63%, and 52.79%; and Day 9 were 25.06%, 20.80%, and 26.47%, respectively. For the mass spawning, the growth rates were 7.836, 7.941, and 7.878 μm/d, respectively. Survival rates at Day 3 were 93.05%, 91.95%, and 92.50%; Day 6 were 79.17%, 78.05%, and 82.50%; and Day 9 were 34.72%, 36.67%, and 38.33%, respectively. The absence of any relationship between shell color and growth and survival at the larval stage may be resulted from their common genetic basis. This work was financially supported by Project of Scientific Innovation, Chinese Academy of Sciences (ZKCX2-211).     

墨西哥湾扇贝稚贝盐度适应性的研究

在室内控制条件下,观察不同海水盐度对墨西哥湾扇贝(Argopecten irradians concentricusSay)稚贝存活及生长的影响,并将168 h 50%死亡的盐度作为适宜生存临界盐度,将增长率为最佳增长率30%时所对应的盐度作为适宜生长临界盐度。结果表明:在稚贝壳高4.38 mm±0.31 mm,水温25.4~27.8℃,pH值7.8~8.2下,墨西哥湾扇贝稚贝适宜生存盐度为23.6~35.7,最适生存盐度为26.0~31.3;适宜生长盐度为23.9~35.8,最适生长盐度为28.7~31.3。在盐度变化超出最适范围时,`墨西哥湾扇贝稚贝对低盐更为敏感,高盐比低盐对稚贝存活、生长、增重的影响小。当盐度为28.7时,墨西哥湾扇贝稚贝具有最高的相对存活率(100%)、壳高日增长率(0.116 2 mm.d-1)及体重日增长率(1.95 mg.d-1)。当盐度在最适范围之外升高或降低,稚贝的存活率、壳高及体重日增长率也随着降低。     

Developmental Dynamics of the Larval Muscle System of Bay Scallop (Argopecten irradians)

Though the larval development of bivalves has been extensively studied for commercial purposes,the dynamic development of larval muscle system remains largely unknown.In this study,we characterized the larval muscle system at different developmental stages(D-shaped veligers,umbo veligers and spats)in the bay scallop(Argopecten irradians)by phalloidin staining and under a confocal microscopy.The functional muscles are initially established at the early stage of veligers,which have four pairs of velar retractors and one anterior adductor.At the veliger stage,the velum and posterior retractor muscles are functionally important for velar contractility but undergo an irreversible shrink until they disappear at the end of the larval stage.During metamorphosis,three crucial modifications take place in the larval muscle system.The metamorphosis process involves the gradual degeneration of velum retractors,mantle margin development from an unfolded to a three-fold state,and remodeling of the adductor muscle system from dimyarian(two adductors)to monomyarian condition(one adductor)as in juveniles/adults.All retractor muscles are composed of striated muscle,but both anterior and posterior adductors have smooth and striated components.These findings highlight that the morphological changes at different stages are typical features of myogenesis in scallops.The present knowledge on the developmental dynamics of myogenesis in the bay scallop will not only improve our understanding of phenotypic diversity of larval myoanatomy in bivalves,but also provide useful information on the larval culture in hatcheries.     

Responses to two-way selection on growth in mass-spawned F1 progeny of Argopecten irradians concentricus (Say)

A bstract In the present study,the effect of one-generation divergent selection on the growth and survival of the bay scallop(Argopecten irradians concentricus)was examined to evaluate the effi cacy of a selection program currently being carried out in Beibu Bay in the South China Sea.A total of 146 adult scallops were randomly selected from the same cultured population of A.i.concentricus,and divided into two groups in shell length(anterior-posterior measurement): large(4.91–6.02 cm,n=74)and small(3.31–4.18 cm,n=72).At the same time,a control group was also randomly sampled(4.21–4.88 cm,n=80).Mass-spawned F 1 progenies from the three size groups were obtained and reared under identical conditions at all growth phases.The effects of two-way(or upward-downward)selection on fertilization rate,hatching rate,survival rate,daily growth in shell length and body weight were assessed in the three size groups.Results show that signifi cant differences(P0.01)were found in hatching rate,survival rate and daily growth of F 1 progenies,but not in fertilization rate(P0.05),among the three groups.The hatching rate,survival rate and daily growth of the progeny of large-sized parents were greater than those of the control group(P0.05),which in turn were larger than those of small-sized group(P0.05).Responses to selection by shell length and body weight were 0.32±0.04 cm and 2.18±0.05 g,respectively,for the upward selection,and-0.14±0.03 cm and-2.77±0.06 g,respectively,for the downward selection.The realized heritability estimates of shell length and body weight were 0.38±0.06 cm and 0.22±0.07 g for the upward selection,and 0.24±0.06 cm and 0.37±0.09 g for the downward selection,respectively.The change in growth by bidirectional selection suggests that high genetic variation may be present in the cultured bay scallop population in China.     

Yield trait improvement of bay scallops following complete diallel crosses between different scallop stocks

This study describes the complete diaUel hybridization between newly introduced bay scallop stock （W） from Canada and local commercial stock （D） grown under laboratory conditions, in China. Larval survival and growth during all life stages （larvae, spat, and adult） were compared among hybrid （DW, WD） and purebred （DD, WW） populations. Significant heterosis was detected for survival during the larval stage （〉 1% of the mid-parent values）. The mean heterosis （Hm） varied in growth throughout the life span. More than 50% of the Hm values were positive and negative in the DW and WD groups, respectively. The influence of maternal effects and mating types （intrapopulation vs. interpopulation crosses） on growth for all life stages was not consistent. Larval survival did not differ significantly （P〉0.05） with maternal effect or mating type. In the harvest stage, shell length （SL）, shell height （SH）, shell width （SW）, and total weight （TW） were larger in the hybrid compared with the inbred groups. Positive Hm values were observed in SL （1.5%）, SW （5.8%）, and TW （12.3%）, and were more significant in the DW groups （6.1%, 4.5%, 6.8%, and 27.2%）. These results suggest that hybridization between two geographic populations is a good tool for improving bay scallop growth. However, unstable heterosis between the two populations requires further study.     

北部湾墨西哥湾扇贝形态性状与闭壳肌质量的关系

为寻找墨西哥湾扇贝中对其闭壳肌重量影响最显著的形态学特征,在整个扇贝的生长过程中,每一个月测一次壳高(mm)、壳宽(mm)、铰合线长(mm)、体质量(g)和闭壳肌质量(g),每一轮的样本采集量为60,通过通径分析研究这些形态学特征对闭壳肌质量的影响。结果显示,根据各形态学特征对闭壳肌质量的直接的、间接的、总路径的贡献,发现体质量的影响是最显著的(p<0.01),其他的形态学特征如壳高、壳宽和铰合线长对于闭壳肌生长既不限制也不促进。     

Effects of elevated seawater pCO<Subscript>2</Subscript> on early development of scallop <Emphasis Type="Italic">Argopecten irradias</Emphasis> (Lamarck, 1819)

To investigate the effects of elevated seawater pCO₂ on the early developmental stages of marine benthic calcifying organisms, we exposed the eggs and larvae of Argopecten irradias, an important bivalve species in Chinese aquaculture, in seawater equilibrated with CO₂-enriched (1000 ppm) gas mixtures. We demonstrated that elevated seawater pCO₂ significantly interfered with fertilization and larval development and resulted in an increased aberration rate. Fertilization in the treatment (pH 7.6) was 74.3% ± 3.8%, which was 9.7% lower than that in the control (pH 8.3) (84.0% ±3.0%). Hatching success decreased by 23.7%, and aberration rate increased by 30.3% under acidic condition. Larvae in acidified seawater still developed a shell during the post-embryonic phase. However, the shell length and height in the treatment were smaller than those in the control. The development of embryos differed significantly at 12 h after fertilization between the two experimental groups. Embryos developed slower in acidified seawater. Nearly half of the embryos in the control developed into D-shaped larvae at 48 h after fertilization, which were considerably more than those in the treatment (11.7%). Results suggest that future ocean acidification (OA) would cause detrimental effects on the early development of A. irradias.     

Effects of Elevated Seawater pCO_2 on Early Development of Scallop Argopecten irradias(Lamarck, 1819)

To investigate the effects of elevated seawater p CO_2 on the early developmental stages of marine benthic calcifying organisms, we exposed the eggs and larvae of Argopecten irradias, an important bivalve species in Chinese aquaculture, in seawater equilibrated with CO_2-enriched(1000 ppm) gas mixtures. We demonstrated that elevated seawater pCO_2 significantly interfered with fertilization and larval development and resulted in an increased aberration rate. Fertilization in the treatment(pH 7.6) was 74.3% ± 3.8%, which was 9.7% lower than that in the control(p H 8.3)(84.0% ±3.0%). Hatching success decreased by 23.7%, and aberration rate increased by 30.3% under acidic condition. Larvae in acidified seawater still developed a shell during the post-embryonic phase. However, the shell length and height in the treatment were smaller than those in the control. The development of embryos differed significantly at 12 h after fertilization between the two experimental groups. Embryos developed slower in acidified seawater. Nearly half of the embryos in the control developed into D-shaped larvae at 48 h after fertilization, which were considerably more than those in the treatment(11.7%). Results suggest that future ocean acidification(OA) would cause detrimental effects on the early development of A. irradias.     

Effects of hypoxia on survival,behavior,and metabolism of Zhikong scallop Chlamys farreri Jones et Preston 1904

Hypoxia,a frequent occurring threat in coastal regions,often results in mass mortalities of marine organisms and brings a serious ecological problem.The commercially important Zhikong scallop Chlamys farreri is being under such a threat as the risks of eutrophication and hypoxia have risen in their culture areas.However,little information has been known concerning their tolerance to hypoxia and their strategy for survival.In the present study,a 20-day experiment was conducted to determine the ef fects of hypoxia on the survival,behavior,and metabolism of Zhikong scallop.With the LC_(50) for dissolved oxygen(DO) being estimated as 1.8 mg/L,the survival of Zhikong scallop can be greatly challenged even under the moderate hypoxic condition of around 2.0 mg/L DO.The survival rate ranged from 69% to 59% when DO dropped from 3.0 to 2.0 mg/L,and it was further reduced to 47% at 1.5 mg/L DO.In hypoxic conditions,the scallops became significantly active,which may be explained as escape attempts to avoid hypoxic water.To save energy,Zhikong scallop would depress their respiration.However,when DO dropped from 3.0 to 2.0 mg/L,the oxygen consumption rate hardly changed.The upregulation of lactate dehydrogenase activity and the unrepressed phosphofructokinase activity,which often result in the unbalanced cellular homeostasis and energy budget,may account for the observed increase in the mortality rate of Zhikong scallops.In general,Zhikong scallop is sensitive to hypoxia events,though possible escape attempts,depressed respiration,and oxaloacetate-pathway may increase their survival chance.     

Effects of different combinations of cryoprotectants on spermatozoon vitality of Chinese scallop,Chlamys farreri,during low temperature equilibrium

The effects of membrane penetrating anti—freezing agents (MPAAs), DMSO (dimethyl sulfoxide), glycerol, EG (ethylene glycol) and methanol in combination with different cryoprotective additives such as carbohydrates, macromoleules and inorganic compounds on the spermatozoon vitality of Chinese scallop,Chlamys farreri, during 1 h 0 °C equilibrium were investigated. When only MPAAs existed, the detrimental effects of different MPAAs ranked in the following order: DMSO<methanol<EG<glycerol. When carbohydrates were added into MPAAs solution, 5% glucose caused larger decrease of spermatozoon vitality than 2.4% lactose. 5% glucose or 2.4% lactose in 7.5% glycerol caused complete damage. 10% yolk was best in maintaining the spermatozoon vitality except when used in combination with 10% methanol. 10% milk significantly decreased spermatozoon vitality in EG and methanol and enhanced its vitality in glycerol, but did not significantly influence it in DMSO. Glycine MPAAs ranked in the following order. 10% yolk <10% yolk +2.4% lactose <2.4% lactose <2.0% citrate <10% milk <10% milk +2.0% citrate <5% glucose <80mmol/L glycine +55 mmol/L NaHCO₃. Contribution No. 2434 from the Institute of Oceanology, Chinese Academy of Sciences.     

Characterization,expression and function analysis of DAX1 gene of scallop (Chlamys farreri jones and preston 1904) during its gametogenesis

DAX1, a member of nuclear receptor superfamily, has a function in the sex determination and gonadal differentiation of several vertebrate species. However, little information about DAX1 of invertebrates is available. Here we cloned a homolog of scallop (Chlamys farreri Jones and Preston 1904) dax1, Cf-dax1, and determined its expression characteristics at mRNA and protein levels. The cDNA sequence of Cf-dax1 was 2093 bp in length, including 1404 bp open reading frame (ORF) encoding 467 amino acids. Unlike those of vertebrates, no conserved LXXLL-related motif was found in the putative DNA binding region of Cf-DAX1. Fluorescence in situ hybridization showed that Cf-dax1 located on the short arm of a pair of subtelocentric chromosomes. Tissue distribution analysis using semi-quantitative RT-PCR revealed that Cf-dax1 expressed widely in adult scallop tissues, with the highest expression level found in adductor muscle, moderate level in mantle, gill and testis, and low level in kidney, ovary and hepatopancreas. The result of quantitative real-time PCR indicated that the expression of Cf-dax1 was significantly higher (P<0.05) in testis than in ovary at the same stage, showing a sex-dimorphic expression pattern. Furthermore, immunohistochemical detection found that Cf-DAX1 mainly located in spermatogonia and spermatocytes of testis and in oogonia and oocytes of ovary, implying that DAX1 may involve in gametogenesis of bivalves.     

Effect of zinc on the growth and development of larvae of bay scallopArgopecten Irradians

The 48-h straight-hinge larvae ofArgopecten irradians were exposed for 9 days to various concentrations of zinc in seawater. The growth rate of larvae in the 50 ppb Zn mediums was 77% that of the controls, and nearly zero in the 200 ppb Zn mediums. A progressive decrease in growth and increase in larval deformity and mortality was observed with increasing zinc concentrations from 50 to 200 ppb. 50 and 100 ppb Zn resulted in delayed development of eye spot and metamorphosis and reduced numbers of larvae at both stages. All larvae subjected to higher zinc concentrations died before attaining the eye-spot stage. The zinc concentrations which caused 50% reduction in growth (G C₅₀) and 50% mortality (LC₅₀) were 109 and 120 ppb respectively. Growing in the medium with 100 ppb Zn, the larvae fed withIsochrysis galbana cultured in a medium with 100 ppb Zn showed more suppressed growth and development than those fed with normal food. It is suggested that zinc contaminated food has strong influence on the bay scallop larvae. Contribution No. 1616 from the Institute of Oceanology, Academia Sinica.     

Survival, growth and immune activity of scallop Chlamys farreri cultured at different depths in Haizhou Bay(Yellow Sea,China)during hot season

Survival, growth and immune response of the scallop, Chlamys farreri, cultured in lantern nets at five different depths (2, 5, 10, 15, and 20 m below the sea surface) were studied in Haizhou Bay during the hot season (summer and autumn) of 2007. Survival and growth rates were quantified bimonthly. Immune activities in hemolymph (superoxide dismutase (SOD) and acid phosphatase (ACP)) were measured to evaluate the health of scallops at the end of the study. Environmental parameters at the five depths were also monitored during the experiment. Mortalities mainly occurred during summer. Survival of scallops suspended at 15 m (78.0%) and 20 m (86.7%) was significantly higher than at 2 m (62.9%), 5 m (60.8%) or 10 m (66.8%) at the end of the study. Mean shell height grew significantly faster at 10 m (205.0 μm/d) and 20 m (236.9 μm/d) than at 2, 5 or 15 m in summer (July 9 to September 1); however, shell growth rate at 20 m was significantly lower than at the other four depths in autumn (September 2 to November 6). In contrast to summer, scallops at 5 m grew faster (262.9 μm/d) during autumn. The growth of soft tissue at different depths showed a similar trend to the shell. Growth rates of shell height and soft tissue were faster in autumn than in summer, with the exception of shell height at 20 m. SOD activity of scallops increased with depth, and ACP activity was significantly higher at 15 and 20 m than at other depths, which suggests that scallops were healthier near the bottom. Factors explaining the depth-related mortality and growth of scallops are also discussed. We conclude that the mass mortality of scallop, C. farreri, during summer can be prevented by moving the culture area to deeper water and yield can be maximized by suspending the scallops in deep water during summer and then transferring them to shallow water in autumn.     

Survival, growth and immune activity of scallop Chlamys farreri cultured at different depths in Haizhou Bay (Yellow Sea, China) during hot season

Survival, growth and immune response of the scallop, Chlamys farreri, cultured in lantern nets at five different depths (2, 5, 10, 15, and 20 m below the sea surface) were studied in Haizhou Bay during the hot season (summer and autumn) of 2007. Survival and growth rates were quantified bimonthly. Immune activities in hemolymph (superoxide dismutase (SOD) and acid phosphatase (ACP)) were measured to evaluate the health of scallops at the end of the study. Environmental parameters at the five depths were also monitored during the experiment. Mortalities mainly occurred during summer. Survival of scallops suspended at 15 m (78.0%) and 20 m (86.7%) was significantly higher than at 2 m (62.9%), 5 m (60.8%) or 10 m (66.8%) at the end of the study. Mean shell height grew significantly faster at 10 m (205.0 μm/d) and 20 m (236.9 μm/d) than at 2, 5 or 15 m in summer (July 9 to September 1); however, shell growth rate at 20 m was significantly lower than at the other four depths in autumn (September 2 to November 6). In contrast to summer, scallops at 5 m grew faster (262.9 μm/d) during autumn. The growth of soft tissue at different depths showed a similar trend to the shell. Growth rates of shell height and soft tissue were faster in autumn than in summer, with the exception of shell height at 20 m. SOD activity of scallops increased with depth, and ACP activity was significantly higher at 15 and 20 m than at other depths, which suggests that scallops were healthier near the bottom. Factors explaining the depth-related mortality and growth of scallops are also discussed. We conclude that the mass mortality of scallop, C. farreri, during summer can be prevented by moving the culture area to deeper water and yield can be maximized by suspending the scallops in deep water during summer and then transferring them to shallow water in autumn.