Mass Selection for Fast Growth in the Third Generation of the Orange Line of the Manila Clam Ruditapes philippinarum

The Manila clam Ruditapes philippinarum is natively distributed along the Pacific coast of Asia, where it is one of the most important bivalve species for local fisheries. This species has been intentionally introduced due to its high adaptability to various coastal environments and is currently widely cultivated in Europe and North America. We have initiated a selective breeding program to improve the growth rate of the Manila clam since 2007, and a full-sib family with orange shell color was selected. This family features high survival but slow growth. Thereafter, two generations of mass selection were conducted in this family by 10% upward selection for faster growth. In 2011, three types of lines were produced by selecting the breeder from the second-generation of selected lines. These lines are SS(subject to three generations of selection for growth in shell length), SC(selected for the second but not the third generation), and C(randomly sampled individuals from the national population). The genetic parameters of the shell length including hR2(realized heritability), SR(selection response), and GG(genetic gain) were analyzed. Results showed that the shell length of the SS line was significantly larger than those of the SC and C lines at all developmental stages(P 0.05). For the larval stage, the values of hR2, SR, and GG were 0.42, 0.73, and 6.66, respectively. For the juvenile stage, the values of hR2, SR, and GG were 0.40, 0.69, 21.76, respectively. For grow-out stage, the values of hR2, SR, and GG were 0.48, 0.83, and 18.22, respectively. The results of hR2 indicated the good potential of the SS line in selective breeding. The level of GG improvement was encouraging and consistent with the previous expectations. No inbreeding depressions in the shell length were observed in the SS and SC lines at all sampling days. All of the results indicate that maintaining selection pressure in successive generations may be effective in the selective breeding program of this family.     

Experimental evaluation of fluorescent(alizarin red S and calcein) and clip-tag markers for stock assessment of ark shell,Anadara broughtonii

Release programs to enhance stocks of ark shell(Anadara broughtonii) have been undertaken in a number of Asian countries,but their effectiveness has rarely been investigated owing to a lack of marking methods.The quality and longevity of fluorescent markers,alizarin red S(ARS) and calcein(CAL)(200 and 300 mg/L),as well as clip tags,were tested on juvenile A.broughtonii.No significant differences in survival or shell growth were observed in juveniles stained with either of the two fluorochromes after a 160-day culture period,but the retention rate was 100%after 1 year.Fluorescent marks(grade 3) were observable microscopically in juveniles stained with the two fluorochromes,and some fluorescent marks(grade 4) were visible with the naked eye after 1 year.ARS-marked shells were brighter than those marked with CAL,and shells marked with 300 mg/L of the fluorochromes were easier to detect than those marked with 200 mg/L.Clip tags were incorporated into the shell as the bivalve grew,and the retention rate was64.25%after 160 days.Significant differences in survival(at 30 days),shell length(at 60,90,120,and 160days),and wet weight(at 90,120,and 160 days) were observed between the clip-tagged and control groups(all P0.05),indicating that the tags may have passive effects on the ark shell.The results suggest that both ARS and CAL are suitable to mark A.broughtonii for large-scale restocking programs,and that optimal marking quality was achieved with 300 mg/L ARS.Lighter and smaller clip tags need to be developed to reduce injury and increase survival rate of clams.     

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.     

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.     

Importance of kelp-derived organic carbon to the scallop <Emphasis Type="Italic">Chlamys farreri</Emphasis> in an integrated multi-trophic aquaculture system

Bivalves and seaweeds are important cleaners that are widely used in integrated multi-trophic aquaculture (IMTA) systems. A beneficial relationship between seaweed and bivalve in the seaweed-based IMTA system has been confirmed, but the trophic importance of seaweed-derived particulate organic materials to the co-cultured bivalve is still unclear. We evaluated the trophic importance of the kelp Saccharina japonica to the co-cultured scallop Chlamys farreri in a typical IMTA farm in Sungo Bay (Weihai, North China). The dynamics of detritus carbon in the water were monitored during the culturing period. The proportion of kelp-derived organic matter in the diet of the co-cultured scallop was assessed via the stable carbon isotope method. Results showed that the detritus carbon in the water ranged from 75.52 to 265.19 μg/L, which was 25.6% to 73.8% of total particulate organic carbon (TPOC) during the study period. The amount of detritus carbon and its proportion in the TPOC changed throughout the culture cycle of the kelp. Stable carbon isotope analysis showed that the cultured scallop obtained 14.1% to 42.8% of its tissue carbon from the kelp, and that the percentages were closely correlated with the proportion of detritus carbon in the water (F =0.993, P= 0.003). Evaluation showed that for 17 000 tons (wet weight) of annual scallop production, the kelp contributed about 139.3 tons of carbon (535.8 tons of dry mass). This confirms that cultured kelp plays a similar trophic role in IMTA systems as it does in a natural kelp bed. It is a major contributor to the detritus pool and supplies a vital food source to filter-feeding scallops in the IMTA system, especially during winter and early spring when phytoplankton are scarce.     

Allelopathic effects of mixotrophic dinoflagellate Akashiwo sanguinea on co-occurring phytoplankton: the significance of nutritional ecology

Blooms of Akashiwo sanguinea frequently break out around the world, causing huge economic losses to the aquaculture industry and seriously damaging coastal ecosystems. However, the formation mechanisms of A. sanguinea blooms remain unclear. We investigated the allelopathic effects of A. sanguinea on multiple phytoplankton species, explored the mode of allelochemicals action and the way of nutrient factors regulation of the allelopathic activity. Results show that strains of A. sanguinea could inhibit the growth of co-occurring phytoplankton including Scrippsiella trochoidea, Phaeocystis globosa, and Rhodomonas salina, but inhibition of Prorocentrum micans was not obvious. The inhibition rates on phytoplankton were positively correlated with the cell densities of A. sanguinea. The highest inhibition rate of 94% on R. salina was for A. sanguinea CCMA256 culture of 2 000 cells/mL at 72 h. We observed that cells of S. trochoidea, Ph. g lobosa, and R. sali na were lysed when co-cultured with A. sanguinea, with the shortest time for S. trochoidea. Additionally, the growth rates of A. sanguinea were promoted by coculturing with S. trochoidea, Ph. globosa, and R. salina. Four components of A. sanguinea culture were all able to inhibit growth of R. salina:the strongest inhibitory effect was found in the sonicated culture, followed by whole-cell culture, filtrates of sonicated culture, and filtrate culture. The crude extract of A. sanguinea culture also lysed cells of R. salina, and the inhibition rates on R. salina increased with the increasing dose of crude extract. It was shown that both nutrient enrichment and nitrogen:phosphorus ratio imbalance enhanced remarkably the allelopathic activity of A. sanguinea. The highest inhibition rate on R. salina of 70% occurred in A. sanguinea JX13 treatment at 2 000 cells/mL under high nutrient condition in 48 h. In JX14 treatment at 2 000 cells/mL for N:P of 10:1, the inhibition rate increased by 1.7 times of that for N:P of 20:1. In addition, the allelopathy of A. sanguinea could not only be a competitive strategy but also a nutrition strategy, playing an important role in formation and/or maintenance of blooms of the mixotrophic dinoflagellate A. sanguinea.     

Use of antioxidant enzymes of clam Ruditapes philippinarum as biomarker to polycyclic aromatic hydrocarbon pollution

The typical organic pollutant polycyclic aromatic hydrocarbon(PAH) anthracene was selected as a contaminant to investigate its eff ects on the activities of superoxide dismutase(SOD), catalase(CAT) and glutathione peroxidase(GSH-Px) in the clam Ruditapes philippinarum. The results show that SOD, CAT and GSH-Px had diff erent induction and inhibition reactions to anthracene stress, and that three diff erent organs in R. philippinarum(visceral mass, muscle tissue and mantle) had diff erent sensitivities to anthracene stress. This study suggest that SOD activities of the visceral mass, CAT activitities of the mantle and the visceral mass, and GSH-Px activity of the muscle tissue could be used as sensitive indicators of anthracene stress in R. philippinarum.     

Introduction,spat-rearing and experimental culture of bay scallop,Argopecten irradians lamarck

Bay scallops were introduced from the east coast of the U.S. into China to shorten the economic turn-over from 2 to 1 year. Parent scallops were carried to Qingdao on Dec. 20, 1982 and stocked in indoor tanks at controlled temperature and fed with a mixture ofPhaeodictylum tricornutum, Pyramimonas sp. andChlorella sp. They spawned on Jan. 26 of the next year. The larvae were reared at a temperature of 18–21°C and fed withIsochrysis galbana, Pyramimonas sp. andChorella sp. In 4 weeks’ growing, the spats averaged 827 μm. They attained a height of 6.9 mm on May 9. In the middle of May, the seed scallops were transferred to Luoyuan Bay in Fujian Province, and Jiaozhou Bay, as well as the area off Taiping Jiao Cape, Qingdao, Shandong Province for experimental culture in plastic netcages suspended on a single line raft. Bay scallops cultured in Luoyuan Bay grew to an averaged shell height of 10.4 mm. In Jiaozhou Bay and the Taiping Jiao Cape culture area, they grew to 50 mm in average shell height (marketable size) and 26 g in average weight by late September; and attained 59 mm in average shell height (R. 39–75 mm) and 46 g in average weight in late December. The ovary and testis could be distinguished by color in August. In early September, eggs and sperms were collected for our laboratory where the second generation of seed scallops was successfully reared to suitable size for growing outdoor and breeding. This species can be harvested within a year after fertilization of the eggs, so we consider it suitable for culture in the Yellow Sea and East China Sea. This is the first reported successful introduction of Atlantic mollusks to the China Seas. This paper was published in Chinese inOceanologia et Limnologia Sinica 17 (5): 367–374, 1986.     

Isozyme Analysis on the Populations of Ruditapes philippinarum

Horizontal starch gel electrophoresis was used to investigate the genetic structure of six populations of Ruditapes philippinarum in the coast of China. Seven enzymes revealed eleven putative loci, and seven of them were polymorphic, PGM^＊, MDH-1^ ＊, MDH-3^＊, LAP-1^＊, LAP-2^＊, MPI-1^＊ and MPI-2^＊. The proportions of polymorphic loci of R. philippinarum populations varied from 0.454 5 to 0.636 4. The values of observed and expected heterozygosities were from 0.039 4 to 0.154 5 and from 0.111 1 to 0.238 5, respectively. Nei＇s genetic distance varied from 0.003 3 to 0.025 3 with an average of 0.013 5. It is suggested that the genetic diversity of R. philippinarum was high.     

Commercial production of tiger puffer (<Emphasis Type="Italic">Takifugu rubripes</Emphasis>) in winter using a recirculating aquaculture system

Tiger puffer (Takifugu rubripes) is a promising species for aquaculture production because of its high value and limited supply. However, in the north of China, using sea cages to culture this species in winter is hampered by the fact that the seawater temperature is extremely low. Here, a large scale commercial production of tiger puffer has been successfully realized using a recirculating aquaculture system (RAS) from 3 October 2012 to 31 May 2013. The RAS was comprised of nine culture tanks (total water volume 200 m³) and stocked with approximately a total of 14400 fish (initial mean weight 160 g). The tiger puffer was hand-fed at a rate of 0.7% of total body weight per day, and the feed conversion rate was (1.21 ± 0.3) kg kg^?1. The recycle water in RAS was treated by a sieve bend screen, a foam fractionator, a submerged biofilter, an UV sterilizer and a submersible aerator. During the whole culture period, an excellent water quality control was achieved in RAS. At the end of this experiment, the survival rate of tiger puffer was more than 98%. The final tank densities averaged 31.2 kg m^?3, and the final individual mean weight was 440 g.     

Relationship Between Shell Color and Growth and Survival Traits in the Pacific Oyster Crassostrea gigas

Shell color is regarded as an economic trait in many breeding programs of bivalve mollusks, but the relationship between shell color and production traits remains controversial. In our breeding program of Crassostrea gigas, solid white, black, and orange shell lines were obtained, and second-generation(F2) and testcross families with segregating shell colors were constructed by crossing these three lines. These segregating families provided ideal samples for investigating the relationship between shell color and production traits in C. gigas. The growth and survival of 7-month-old oysters with different shell colors sampled within the same families were compared in seven F2 families and 13 testcrosses. In addition, the growth and survival of oysters from the three shell color lines were compared at 4 and 16 months of age. The growth and survival rates of the orange shell line were significantly lower than those of the white and black shell lines. However, no significant difference in growth between oysters with different shell colors was observed within segregating families, except the testcrosses produced by crossing orange-and white-shelled grandparents, and no significant difference in survival was observed in any family. Overall, no significant correlation was observed between shell color and production traits in C. gigas. These results suggest that shell color cannot be used as a marker to guide the selection of growth and survival traits. Thus shell color and production traits should be selected independently in oyster breeding programs.     

Effects of starving and re-feeding strategies on the growth performance and physiological characteristics of the juvenile tongue sole (<Emphasis Type="Italic">Cynoglossus semilaevis</Emphasis>)

Starving and re-feeding trials were conducted to evaluate the effects of starvation duration and recovery times on the growth performance and physiological characteristics of juvenile tongue sole Cynoglossus semilaevis, which included growth, body composition, intestinal morphology and digestive enzymes activities. The fish starved for one (D1), two (D2) and four (D4) days, respectively, were recovered for three (R3, R6, R12), seven (R7, R14, R28) and eleven (R11, R22, R44) folds of starvation days while the control fish were fed constantly. The experiment was conducted for 96 days. The fish individuals in D1R11, D2R14 and D2R22 caught up in weight with the control, indicating that complete compensatory growth existed in these fish individuals. The highest body weight gain and total specific growth rates were observed in D2R22. Food intake increased significantly in most treatments in comparison with that of the control except for D1R7 and D2R22 (P < 0.05). Food conversion efficiency and apparent digest coefficient in D2R22 were significantly promoted, being higher than that of the control (P < 0.05). The activities of trypsin and lipase were found to be closely related to the growth performance. The activities of liver and hindgut trypsin, also midgut and hindgut lipase in D2R22 were significantly higher than those of the control (P < 0.05). The fold height (HF) of foregut and midgut increased significantly in D2R22, and HF decreased significantly in D1R3, D2R6 and D4R12 (P < 0.05). On our findings, we may conclude that the optimum starving and re-feeding strategy is starving for 2 days and re-feeding for 22 days. And in this starving and re-feeding strategy, the compensatory growth could be mostly attributed to the promotion of food conversion efficiency and digestibility coefficiency.     

Relationships of growth and mortality to enzymatic activity,and the relative mRNA expression of cultured scallops Patinopecten yessoensis in the Yellow Sea,China

The bottom aquaculture of yesso scallop(Pa tinopecten yessoensis)has developed rapidly in the north of Yellow Sea,China,but not without accompanying productivity problems.We,therefore,conducted field surveys to investigate factors related to growth and mortality rates of bottom-cultured scallops after release.Specifically,we focused on the effects of total antioxidant capacity(TAOC)and three key enzymes involved in antioxidant and metabolic function:glutamic pyruvic transaminase(GPT),lactic dehydrogenase(LDH),and superoxide dismutase(SOD).Across summer and winter,we also measured the relative mRNA expression of GPT and SOD to understand their seasonal variation in the scallops,along with how such variation correlated to growth and mortality.Results show that bottom cultured scallops experienced mass mortality during the first six months post-release.During winter(December),scallops grew more rapidly and suffered less mortality than in summer(August).The observed lower performance probably resulted from less advantageous environmental factors during the summer seasons,such as high temperature and low dissolved oxygen.These environmental stressors enhance protein consumption while decreasing energetic resources in scallops.Furthermore,scallops in summer exhibited high antioxidant levels that probably competed for energy with process integral to growth and survival.These negative factors combined to elevate mortality rates.In conclusion,we provided evidence suggesting correlations between metabolic/antioxidant activity and growth and mortality of bottom-cultured yesso scallops.These correlations implied us an accurate method to estimate the performance of bottom culture system.Suggestions about innovative aquaculture techniques were also discussed in the study.Our results might provide a possible guideline to the improvement of bottom culture techniques for this commercially valuable seafood species.     

Ontogenetic development in the morphology and behavior of loach （Misgurnus anguillicaudatus） during early life stages

Loach （Misgurnus anguillicaudatus） is a commercially important fish in China and an ideal aquaculture species. However, culturists experience high larval and juvenile mortality during mass production. To provide insight into ways to improve larviculture techniques, we describe the morphological characteristics and behavior of loach during the larval and early juvenile stages. Yolksae larvae ranged from 2.8 to 4.0 mm body length （BL） between days 0 to 4; preflexion larvae ranged from 3.6 to 5.5 mm BL between days 4 to 6; flexion larvae ranged from 4.8 to 8.1 mm BL between days 5 and 14; and post-flexion larvae ranged from 7. l to 15.7 mm BL between days 11 to 27; the minimum length and age of juveniles was 14.1 mm BL and 23 d, respectively. Loach are demersal from hatch through to the early juvenile stages. A suite of morphological characteristics （e.g., external gill filament and ventral mouth opening） and behavioral traits have developed to adapt to demersal living. We observed positive allometric growth in eye diameter, head length, head height, and pectoral fin length during the early larval stages, reflecting the priorities in the development of the organs essential for survival. Our results provide a basis for developing techniques to improve the survival of larval and juvenile loach during mass production.     

菊黄东方[鱼屯]1龄幼鱼生长特性

对600尾菊黄东方[鱼屯]（Takfugu flavidus） 1龄幼鱼在池塘养殖条件下的生长特性进行研究。结果表明;菊 黄东方[鱼屯]67日龄幼鱼经过127d的培育后,体长从（ 3.53±0.73）cm增至（ 10.33±0.98） cm,日均增长0.054 cm, 体长增长率193.14%;体质量从（2.41± 1.60）g增至（ 54.39±13.62 ） g,日均增重0.413 g,体质量增长率2135.8%, 体质量与体长呈幕函数关系,关系式为：W=0.0448L30570 （R2=0.9922）, b值接近于3,属等速生长型。体长 （l）与日龄（t）表现为线性相关：l= 0.0585t ＋ 3.7811 （R2= 0.9621 ）,体质量（w）与日龄（t）表现为指数相 关：w=3.3211e00286t （R2=0.9363）。     

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).     

Morphological and karyotypic variation in three wild populations of Meretrix meretrix

Three wild populations of Meretrix meretrix sampled from Dongxing, Beihai, and Shankou along the coast of Guangxi, China, were investigated with morphometry and karyometry. Six morphological indices （shell length, shell height, shell width, hinge length, total wet weight and shell weight） were measured. Differences in all morphological indices except hinge length were significant among the three populations （P 〈 0.05）. The mean values of these indices （except for the hinge length） in the Dongxing population were larger than those in the Beihai and Shankou populations, although the latter had the largest hinge length. The karyotype of the Beihai, Shankou and Dongxing samples had ten metacentric, six submetacentric, and three subtelocentric chromosome pairs. No significant difference was shown in the centromeric index values of the chromosomes in the populations （P〉0.05）. However, the order of metacentric, submetacentric and subtelocentric chromosome pairs was variable among the three populations. The results indicate a high level of inter-population variation in morphology and karyotype.     

Application of Wet Waste from Shrimp (Litopenaeus vannamei) with or Without Sea Mud to Feeding Sea Cucumber (Stichopus monotuberculatus)

In the present study, the applicability of the wet waste collected from shrimp(Litopenaeus vannamei) to the culture of sea cucumber(Stichopus monotuberculatus) was determined. The effects of dietary wet shrimp waste on the survival, specific growth rate(SGR), fecal production rate(FPR), ammonia- and nitrite-nitrogen productions of sea cucumber were studied. The total organic matter(TOM) level in the feces of sea cucumber was compared with that in corresponding feeds. Diet C(50% wet shrimp waste and 50% sea mud mash) made sea cucumber grow faster than other diets. Sea cucumber fed with either diet D(25% wet shrimp waste and 75% sea mud mash) or sole sea mud exhibited negative growth. The average lowest total FPR of sea cucumber occurred in diet A(wet shrimp waste), and there was no significant difference in total FPR between diet C and diet E(sea mud mash)(P > 0.05). The average ammonia-nitrogen production of sea cucumber in different diet treatments decreased gradually with the decrease of crude protein content in different diets. The average highest nitrite-nitrogen production occurred in diet E treatment, and there was no significant difference in nitrite-nitrogen production among diet A, diet B(75% wet shrimp waste and 25% sea mud mash) and diet C treatments(P > 0.05). In each diet treatment, the total organic matter(TOM) level in feces decreased to different extent compared with that in corresponding feeds.