Induced nest spawning and artificial hatching of the fertilized eggs of mudskipper, Boleophthalmus pectinirostris

In this study, nest spawning was successfully induced by exogenous hormone injections and seawater flow stimulation, and optimum condition for hatching fertilized eggs of burrow fish mudskipper,Boleophthalmus pectinirostris, was searched. Apart from spawning inside the nests, females also spawned outside the nests. The percentages of spawned nests were 8.0% to 24.2%. Most eggs were observed adhered to the inner wall of the top half of the nest. Fertilization rates of the nest-spawned eggs varied from 17.3% to 80.8% Females could spawn after being artificially confined inside the‘nests with males at ratios of 1:1 or 1:2, but the spawned eggs were not fertilized. Mean hatching rates of artificially fertilized eggs incubated in round plastic buckets were 32.7%-70.6%, and in the net cages, were 4.2%-20.5%, respectively. Mean hatching rates of nestfertilized eggs incubated in the round plastic buckets were 33.6%-76.3%, and in the net cages, were 5.9%-25.2%.Results showed that round bucket incubation was the best way for hatching fertilized eggs of mudskipper.Keeping the hatching seawater flowing is an important way for increasing the hatching rates of the mudskipper fertilized eggs.     

BIOCHEMICAL CHANGES DURING DEVELOPMENT OF EGGS AND YOLK-SAC LARVAE OF HERRING AND PLAICE

Changes in quantities of water, sodium, potassium, lipid and protein were compared in developing eggs and yolk-sac larvae of herring dupes harengus L., a species with demersal eggs, and plaice Pleuroncctcs platessa L., a species with pelagic eggs. The principal difference between the two species was the occurrence in herring, but not in plaice of considerable increases in water content and in one or both of the above ions at three distinct stages: at fertilization,during the middle stage of egg development, and in the days immediately after hatching. Newly hatched larvae of the two species differed greatly in water content because of the difference in mass and water content of the yolk, not in the water content of the rest of the body. In herring, there were increases in sodium and potassium which coincided broadly with the increase in water content. These differences are related to the gradual acquisition of buoyancy by herring during this period, comparable with the buoyancy conferred on plaice eggs a     

EFFECT OF ZINC ON THE GROWTH AND DEVELOPMENT OF LARVAE OF BAY SCALLOP ARGOPECTEN IRRADIANS

The 48-h straight-hinge larvae of Argopecten 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 (LC 50)were 109 and 120 ppb respectively. Growing in the medium with 100 ppb Zn, the larvae fed with Isochysis galbana cultured in a medium with 100 ppb Zn showed more suppressed growth and development than those fed with nonnal food. It is suggested that zinc contaminated food has strong influence on the bay sca     

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.     

Feeding Broodstocks Different Starfish Diets Affect Growth and Survival of Larvae of Trumpet Shell(Charonia lampas sauliae Reeve 1844)

Trumpet shell(Charonia lampas sauliae)(Mollusca, Heterogastropoda, Cymatidae) has extensive economic value. Studies on the artificial larval development of C. lampas sauliae for aquaculture utilization have become especially important due to the finite natural resources. In the present study, the growth and survival rate of the larvae of C. lampas sauliae broodstocks fed three types of starfish diets, Asterina pectinifera Müller Troschel 1842, A. amurensis Lütken 1871 and their mixture were compared. The larval size increased gradually between day 10 and day 20 after hatching at 15℃ and 20℃. No difference was found in body size and specific growth rate(SGR)(two-way ANOVA; P 0.05). However, during transition from trochophore to veliger stage 20 days after hatching, significant increases in larval survival and growth rates were observed. The maximum survival rate was observed on day 10. The mean survival rate was 0.463, 0.730 and 0.515 at 15℃, and 0.369, 0.713 and 0.444 at 20 when ℃ A. pectinifera, A. amurensis and their mixture were fed, respectively. The SGR and survival rate of the larvae were definitely influenced by the diets(P 0.05), and the effect of A. amurensis alone was higher than that of A. pectinifera alone and their mixture.     

Otolith development in larval and juvenile Schizothorax davidi: ontogeny and growth increment characteristics

Laboratory-reared Schizothorax davidi larvae and juveniles were examined to assess the formation and characteristics of David’s schizothoracin otoliths.Otolith development was observed and their formation period was verifi ed by monitoring larvae and juveniles of knownage.The results revealed that lapilli and sagittae developed before hatching,and the first otolith increment was identified at 2 days post hatching in both.The shape of lapilli was relatively stable during development compared with that of sagittae;however,growth of four sagittae and lapilli areas was consistent,but the posterior area grew faster than the anterior area and the ventral surface grew faster than the dorsal surface.Similarly,the sum length of the radius of the anterior and posterior areas on sagittae and lapilli were linearly and binomially related to total fish length,respectively.Moreover,daily deposition rates were validated by monitoring knownage larvae and juveniles.The increase in lapilli width was 1.88±0.080 0μm at the ninth increment,which reached a maximum and the decreased gradually toward the otolith edge,whereas that of sagittae increased more slowly.These results illustrate the developmental biology of S.davidi,which will aid in population conservation and fish stock management.     

Early development and the point of no return in pikeperch( Sander lucioperca L.) larvae

The objectives of the present study were to evaluate the yolk-sac and oil globule absorption and point of no return(PNR) of pikeperch(S ander lucioperca L.) larvae. Artificial propagation of pikeperch was performed at(15±2)°C. Yolk-sac absorption, oil globule absorption, larval growth and the first initial feeding rate were observed to analyze the early growth and to determine the PNR of pikeperch larvae. The total length of newly hatched(0 day after hatching, DAH) pikeperch larvae was(4.25±0.22) mm and the volume of the yolk-sac and the oil globule was(0.30±0.12) mm3 and(5.14±2.28) 10-2 mm 3 respectively. The yolk-sac and the oil globule were exhausted at 11 DAH and 14 DAH, respectively. Pikeperch larvae began feeding at 8 DAH with an initial feeding rate about 10.0%. From 9 to 14 DAH, the initial feeding rate increased rapidly, and reached its highest(about 82.7%) at 14 DAH. It declined thereafter, 48.9% at 15 DAH and 35.6% at 16 DAH, thus the pikeperch larvae reached PNR by 15–16 DAH. The appropriate first feeding time for the pikeperch larvae is 11–12 DAH, when the initial feeding rate is higher than half of the maximum initial feeding rate.     

Influence of salinity on the early development and biochemical dynamics of a marine fish,Inimicus japonicus

Fertilised eggs of the devil stringer(I nimicus japonicus) were incubated at different salinity levels(21, 25, 29, 33, and 37), and then the hatching performances, morphological parameters, and biochemical composition(protein, lipid and carbohydrate) of the larvae were assayed to determine the influence of salinity on the early development of I. japonicus. The tested salinity levels did not af fect the times of hatching or mouth opening for yolk-sac larvae. However, the salinity significantly influenced the hatching and survival rates of open-mouthed larvae, as well as the morphology of yolk-sac larvae. The data indicated that 30.5 to 37.3 and 24.4 to 29.8 were suitable salinity ranges for the survival of embryos and larvae of I. japonicus, respectively. Larvae incubated at a salinity level of 29 had the greatest full lengths, and decreasing yolk volume was positively correlated with the environmental salinity. With increasing salinity, the individual dry weights of newly hatched larvae or open-mouthed larvae decreased significantly. Newly hatched larvae incubated at a salinity level of 29 had the greatest metabolic substrate contents and gross energy levels, while the openmouthed larvae's greatest values occurred at a salinity level of 25. Larvae incubated in the salinity range of 33 to 37 had the lowest nutritional reserves and energy values. Thus, the I. japonicus yolk-sac larvae acclimated more readily to the lower salinity level than the embryos, and higher salinity levels negatively influenced larval growth and development. In conclusion, the environmental salinity level should be maintained at 29–33 during embryogenesis and at 25–29 during early larval development for this species. Our results can be used to provide optimum aquaculture conditions for the early larval development of I. japonicus.     

Polyploidy induction by hydrostatic pressure shock and embryo development of sea cucumber Apostichopus japonicus

The manipulation of the chromosome set for commercially valuable marine animals is important for enhancing aquacultural production. In this study, triploid and tetraploid sea cucumber Apostichopus japonicus were induced by hydrostatic pressure shock, and the conditions of appropriate induction were tested with different starting times, and hydrostatic pressure intensities and durations. The highest rate of triploid induction reached 20% and that of tetraploid was 60%. In consideration of the survival rate and hatch rate, the appropriate treatment for triploid was 55 Mpa of hydrostatic pressure for 5 min at 55 min after fertilization (a.f.), while for tetraploid it was 60 Mpa for 5 min at 61 min a.f. The triploid of the sea cucumber could survive through the pelagic larval stage and attachment stage, and develop like the control group of the experiment. The tetraploid, however, could not survive the attachment stage.     

Inbreeding Depression on Growth and Survival of Full-Sib Family of Manila Clam(Ruditapes philippinarum)

In present study, the inbreeding depression(ID) of growth and survival of Manila clam(Ruditapes philippinarum) was investigated at larval and juvenile stages. Nine inbred families(A_2, B_2, C_2, D_2, E_2, F_2, G_2, H_2 and I_2) were established by mating within nine full-sib families with expected inbreeding coefficient of 0.25. Inbred families showed significant differences in shell length and hatching rate of D-larvae(straight-hinged larvae). The larvae of the nine inbred families grew slower than those of control group(CG), and their ID value ranged from 0.81% ± 6.09% to 16.10% ± 1.49%. The ID value of larval survival rate varied between 27.47% ± 9.36% and 70.50% ± 13.66%. The ID was also detected for juvenile growth in A_2, B_2, C_2, and D_2, which ranged from 4.60 ± 2.21 to 17.71 ± 7.73. The A_2 family maintained the highest juvenile survival rate, whereas the other inbred families exhibited ID values varying between 62.79% ± 4.54% and 96.14% ± 0.87%. The linear relationship of estimated ID between growth and survival was negatively correlated(R =-0.434, P 0.05). The results of this study suggested that the ID of growth was common at the larval stage but was less prevalent at juvenile stage. In contrast, the ID of survival increased from larval to juvenile stage. A better understanding of the effect of inbreeding may aid to selective breeding of Manila clam.     

Induced nest spawning and artificial hatching of the fertilized eggs of mudskipper, Boleophthalmus pectinirostris

In this study, nest spawning was successfully induced by exogenous hormone injections and seawater flow stimulation, and optimum condition for hatching fertilized eggs of burrow fish mudskipper,Boleophthalmus pectinirostris, was searched. Apart from spawning inside the nests, females also spawned outside the nests. The percentages of spawned nests were 8.0% to 24.2%. Most eggs were observed adhered to the inner wall of the top half of the nest. Fertilization rates of the nest-spawned eggs varied from 17.3% to 80.8%. Females could spawn after being artificially confined inside the nests with males at ratios of 1∶1 or 1∶2, but the spawned eggs were not fertilized. Mean hatching rates of artificially fertilized eggs incubated in round plastic buckets were 32.7%–70.6%, and in the net cages, were 4.2%–20.5%, respectively. Mean hatching rates of nest-fertilized eggs incubated in the round plastic buckets were 33.6%–76.3%, and in the net cages, were 5.9%–25.2%. Results showed that round bucket incubation was the best way for hatching fertilized eggs of mudskipper. Keeping the hatching seawater flowing is an important way for increasing the hatching rates of the mudskipper fertilized eggs. Supported by the NSFC (No. 30170739), and also supported by the Provincial Natural Science Foundation of Fujian Province (No. B0210003).     

Triploidy induction with heat shocks toPenaeus chinensis and their effects on gonad development

Heat shocks effectively produced triploids inPenaeus chinensis. Fertilized eggs heat shocked (28–32°C) for 8 to 16 minutes, starting from 8 to 20 minutes after spawning, resulted in triploidy induction rates of 39%–75%. Several triploid populations were cultured to 10 cm. In a triploid population, two kinds of ovaries were observed. Histological examination showed apparent differences between these two kinds of ovaries, whereas among male shrimp, there were no such differences. Contribution No. 3465 from the Institute of Oceanology. Chinese Academy of Sciences. The study was supported by the International Foundation for Science, Asian Fisheries Society and Chinese Climbing Project PDB 6–2, and Chinese National 863 Project (No. 863-819-01-05).     

Tetraploid induction by inhibiting mitosis I in scallopChlamys farreri

Tetraploid induction was carried out by inhibiting mitosis I in fertilized eggs ofChlamys farreri. Mitosis I was blocked with cold shock (5–7°C), Cytochalasin B (0.75 mg/L) and 6-dimethylaminopurine (6-DMAP) (60–75 mg/L) when 60% fertilized eggs released polar body II at 20°C. At 4-cells embryo stage, the ploidy was determined by counting chromosome number. In control groups, most embryos were diploids (72.22%) and aneuploids (24.78%). In Cytochalasin B, cold shock and 6-DMAP treated groups, tetraploids were respectively 10.51%, 4.08%, and 13.34%; aneuploids were 43.10%, 35.93% and 29.16%, and triploids were 7.84%, 8.52% and 18.33%. At D-larva stage, ploidy was determined by flow cytometry (FCM). The ploidy analysis of day 2 larvae showed diploids in control group and also in three treated groups. Juvenile scallops (0.2–0.3cm) which were harvested in two control groups and two CB treated groups were all diploids through checking ploidy individually by FCM. Contribution No. 3648 from the Institute of Oceanology, Chinese Academy of Sciences. Contribution No. 238 from the Experimental Marine Biology Laboratory, Chinese Academy of Sciences. Project funded by grant (No. 819-01-07) from the Chinese Science and Technology Ministry and also by “Hundred Talents Plan” grant from the Chinese Academy of Sciences.     

Production of bioplastics and hydrogen gas by photosynthetic microorganisms

Experiments on salimity tolerance of embryos and larvae of the southern bay scallopArgopecten irradians concentricus Say under controlled temperature of 28°C showed that although the eggs were fertilized and cleaved at salinity 18.1–37.4, they could further develop into normal D-shaped larvae only at salinity 23.4–36.1, while the trochophore larvae could develop into normal D—shaped larvae at salinity 22.1–39.9. For 72 hours, the larvae could tolerate salinity of 18.4–43.9, the salinity favoring growth was 19.8–42.4; the suitable range was 21–36, and the optimum at about 28. They metamorphosed at salinity 18.9–43.5, optimally at 24–32. Contribution No. 3301 from the Institute of Oceanology, Chinese Academy of Sciences. The research was supported by the National Climbing Project B(PDB6-3-2).     

Triploidy induction in Pacific oysterCrassostrea gigas by caffeine with thermal shock

Zygotes of Pacific oyster (Crassostrea gigas) were treated for triploid induction with caffeine (10 mmol/L, 15mmol/L and 20mmol/L) in combination with thermal shocks (at 40 minute post-fertilization) lasting for 5 and 10 minutes. The highest yield of triploids, 41.5%, was obtained from the treatment with 20 mmol/L caffeine at 34°C lasting 10 minutes. The triploid levels were less than 30.0% in other treatments. Triploid induction was more effective in treatment with 15–20 mmol/L caffeine at 34–38 °C than with lower concentrations of caffeine at temperatures below 32 °C. Our results suggest that triploid induction with caffeine in combination with thermal shocks is less efficient than some other methods reported previously.     

Comparison of taste components between triploid and diploid oyster

The main taste components of triploid and diploid oyster (Crassostrea gigas) were compared. Free amino acids, inosine monophosphate, succinate, trimethylamine oxide and betaine in fresh and boiled extractives were analyzed. The protein, lipids, glycogen, moisture and ash, which may affect the flavour, were evaluated. In boiled extractives, the amino acids were 394.1 mg(100 g)⁻¹ in diploid and 183.5 mg(100 g)⁻¹ in triploid. However, in fresh oyster extractives, they were 320.0 mg(100g)⁻¹ and 147.3 mg(100g)⁻¹ respectively. The inosine monophosphate in triploid was 44% more than that in diploid, and a little difference existed in the content of trimethylamine oxide between them. The contents of these taste components were the basis for taste flavour pattern determination.     

鲫鲤杂交一代（F_1）自交的受精细胞学观察

采用人工催产、波恩氏液固定、石蜡切片和Ｈ·Ｅ染色等方法，研究了杂交一代（Ｆ１）自交的受精细胞学过程。结果表明，鲫鲤杂交一代自交时有正常的受精细胞学过程。受精后，卵子完成第二次成熟分裂，排出第二极体；１５ｍｉｎ，雌雄性原核形成，２０ｍｉｎ，两性原核完全融合，形成二倍体的合子，４０ｍｉｎ，开始进行卵裂；１ｈ，两个分裂球出现，本实验为Ｆ２代是二倍体提供了证据。     

AVAILABILITY OF PHOSPHORUS FROM SELECTED INORGANIC PHOSPHATES TO JUVENILE ABALONE, HALIOTIS DISCUS HANNAI INO

The availability of phosphorus to juvenile abalone,Haliotis discus hannai from primary, secondary and tertiary calcium phosphate, primary sodium or potassium phosphate separately or in combination was determined in a 120-day feeding trial. Seven semi-purified diets were formulated to contain relatively constant dietary phosphorus, ranging from 0.78%–0.82%. The concentrations of dietary phosphorus were presumed to be slightly below the requirement for abalone. A brown alga,Laminaria japonica, was used as a control diet. Abalone juveniles of silimar size (average weight 1.18 g; average shell length 18.74 mm) were distributed in a single-pass, flow-through system using a completely randomized design with eight treatments and three replicates each treatment. The survival ranged from 71.1% to 81.1%, and was not significantly (P>0.05) related to dietary treatment. However, the weight gain rate (41.72%–65.02%), daily increment in shell length (36.87–55.07 μm) and muscle RNA-DNA ratio (3.44–4.69) were significantly (P<0.05) affected by dietary treatment. Soft body alkaline phosphatase activity (10.9–19.8 U/g wet tissue) and carcass levels of lipid (7.71%–9.33%) and protein (46.68%–49.35%) were significantly (P<0.05) responsive to available phosphorus of the diets. Dietary treatment had significant effect (P<0.05) on concentrations of phosphorus in the whole body (WB) and soft body (SB). Apparent digestibility coefficients (45%–97%) of phosphorus were also significantly (P<0.05) different between dietary treatments. The results indicated that among these inorganic compounds, primary calcium, potassium and sodium phosphates separately or in combination could be utilized effectively by juvenile abalone as dietary phosphorus sources (availability ranging from 87%–97%). However, secondary and tertiary calcium phosphates were found to be low in availability, the values being 45% and 77%, repectively. Project 39670572 supported by the Natural Science Foundation of China (NSFC).     

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).     

Growth,reproduction & population structure of the freshwater crabSinopotamon yangtsekiense bott, 1967, from Zhejiang,China

Monthly investigations were mae on the population of Chinese freshwater crab,Sinopotamon yangtsekiense Bott, 1967 from April, 1984 to March, 1985. The data on 4413 specimens show that the growth was affected mainly by temperature. During the April to November growth period, the crabs' major development occurred from June through October. One year was required for a fine white oocyte to develop into a mature egg. The reproduction period was June–October. Females bearing eggs were taken from June–August, and crabs with young were found from July–October. The females reproduced once a year but could for more than one year. The number of eggs carried by a female varied greatly according to the size of the crab, ranging from 30 to 100 eggs. New-born crabs become mature after 1–2 years. The sex ratio was approximately 1∶1 in the overall population. However, the larger crabs are predominantly male. The age distribution ofS. yangtsekinese was estimated from size frequency histograms. There were more adult crabs (over 70%) from June to October and more immature crabs (over 50%) from November to May.