Abstract:The embryonic development of golden cuttlefish under artificial hatching conditions was observed. The average weight of newly spawned eggs is 0.99 g/particle (n=100), and the major axis is 11~24 mm, the minor axis is 8~15 mm. Egg yolk is egg-shaped, pear-shaped, with long diameter of 5~6 mm and short diameter of 3~4 mm, and is wrapped by three-egg membrane outside. Three egg membrane in the embryonic development processed a soft figuration narrow enlargement change. The fertilized eggs hatched at water temperature of 20~25℃, and the larvae hatched out about after 26d hatching. In terms of the exterior appearance and the typical characteristics, process of embryonic development includes 5 stages: cleavage stage, blastula stage, gastrulae stage, organ development stage and hatching stage. In the embryonic development process, the morphological changes of the eye were relatively large, and it turned from colorless, red into black. KEY WORDSGolden cuttlefish(Sepia esculenta Hoyle)Embryonic development

Abstract:The ultrastructure of barfin flounder Verasper moseri spermatozoon was studied with scanning microscopy and trans mission electron microscopy. Total length of the sperm is 45.57±3.23μm (n=11), and the sperm includes three parts: head, mid piece and flagellum. The spherical head mainly consists of the nearly circular cell nucleus. The mid piece is composed of mitochondria and center grain complex, with some round undifferentiated small mitochondria. The typical ‘9+2’ pattern of the flagellum has quite simple structure, which means that the spermatozoon of barfin flounder can be classified as a primitive type, which is found in most of the advanced fish taxa. But the anterior vesicles and pit of the nucleus of this spermatozoon have seldom reported from other species. There is no acrosome in the sperm. At anterolateral to the nucleus, there is a chromatin free, electron lucent area, which was named the anterior pit of the nucleus. Within and in front of the pit, an irregular shaped structure and some different sized vesicles that are encapsulated in a layer of membrane were found. With regard to acrosome and acrosome vestige present in some other euteleosts, we presume that the vesicles may represent acrosome vestiges though they may also function to release nuclear material, while the irregular shaped structure in front of the nucleus may be caused by improper sample preparation

Abstract:Ten meristic traits and seven morphological characteristics of marbled sole, Pleuronectes yokohamae (78 individuals), collected from Penglai, North Korea and Kamaishi populations, were measured and compared by the principal component analysis method, multivariate analysis method and one way ANOVA. The multivariate analysis and one way ANOVA results show that there are significant differences (P=0.05) among the three populations in two to five morphological characteristics. Discriminant analysis on the high contributing morphological parameters show that there are extremely significant differences among the three populations of P. yokohamae, with 83.3%～100.0% identification accuracy and 92.31% synthetic identification accuracy. There are significant differences between Chinese population and Japanese population, and a certain extent of geographical variances existed in the three populations

Abstract:The types and distribution of mucous cells in the alimentary canal of Cheilinus undulates Rüppell were studied using the AB-PAS (Alcian blue and periodic acid Schiff's reagent) staining method. The mucous cells are distributed in every part of the alimentary canal of Cheilinus undulates Rüppell, however, the cell densities and cell types vary at different locations. Type Ⅱ mucous cells are mainly located near the gastrovascular cavity, while type Ⅲ and type Ⅳ mucous cells are mainly found in the bottom of the mucous layer. Buccopharyngeal cavity, tongue and esophagus mainly posesse type Ⅱ and type Ⅳ mucous cells. The top of protuberances in small intestine and recta is characterized by type Ⅱ and type Ⅳ mucous cells, and the bottom is characterized by type Ⅲ and type Ⅳ mucous cells.KEY WORDSCheilinus undulates RüppellDigestive tractMucous cellsTypes and distribution

Abstract:Nutrient components in the muscle of Humphead wrasse Cheilinus undulates were investigated in this paper. The results are as follows: the content of meat in whole fish was 71.5%, and the crude protein, crude fat, crude ash and moisture in fresh muscle of Humphead wrasse were 18.98 %, 1.56 %, 1.23 % and 76.23 %, respectively. Eighteen common amino acids were found in the muscle of fish and the total content of amino acids ( TAA) was 878.1mg/g (dry weight). Four kinds of delicious amino acids (DAA) were accounted for 355.3 mg/g (dry weight), and this is the main factor that the fish is delicious. The contents of EPA and DHA in fatty acids were 2.0 % and 9.51 %, respectively , EPA+DHA were 12.76% of the total fatty acids.

Abstract:Myostatin is a member of the transforming growth factor-β superfamily, which functions as a negative regulator for the development and growth of skeletal muscle in animal. Based on the cloned myostatin-1 gene of Larimichthys crocea, the promoter sequence of L. crocea myostatin11 gene was cloned by genomic walking. The sequencing results indicate that it shares 90% and 75% homology identity with the corresponding reported sequences of Sparus aurata and Micropterus almoides, respectively. Bioinformatic analysis showed that myostatin-1 gene contains one consensus sequences of TATA box, a CAAT box and five putative E-boxes known as the binding sites to myogenic basic helix-loop helix transcription factors. In addition, it also possesses many transcription factor binding sites, such as MyoD, USF, MEF2, SP1 and NF-Y. These results are expected to be the basis for further studies on the regulation and expression of this gene.KEY WORDSLarimichthys croceaMyostatin-1 promoterCloningBioinformatic analysis

Abstract:Amphiprion ocellaris, an ornamental fish collected from the wild, was cultured to mature and spawned in artificial condition. The embryonic development was observed under a microscope and it was found that the development was similar to most teleosts. The eggs belong to the telolecithal and are subject to discoidal cleavage. The incubation period was 192~216 hours at 27.0±0.5℃. The larval fish could be fed by rotifer in the second morning after hatching, and were developed into juvenile at 4~5 days after hatching and into parr at 18~20 days after hatching. The parrs could be fed by artificial food and shrimp meat at 45~50 days after hatching.

Abstract:Triploid induction of Argopectens irradias (Lamarck) was achieved through inhibiting the second polar body release by soaking fertilized eggs in sea water with low salinity, and the effect of different salinity treatment on triploid induction was examined. When 40％ eggs reached the first polar body stage, the eggs were soaked in sea water with salinities of 30, 22.5, 20, 17.5, 15, 12.5, 10 and 7.5 for 10 min or 15 min respectively. The eggs were then transferred into the normal salinity seawater for 24h hatching. Triploid rate was no more than 2.91% when the salinity was above 20. However, with the decrease of salinity, triploid induction was enhanced so dramatically that the rate of triploid reached up to 85.91％ at salinity of 12.5. Though, when the salinity was below 7.5, due to the lack of D-shaped larvae, the triploid rate could not be measured. The results show that the induction effect of 15 min treatment is better than 10 min. The experiment results suggest that the best triploid induction of A. irradias is achieved by bathing in seawater at salinities of 12.5~15 for 15 min.KEY WORDSArgopectens irradiasLow salinityTriploidD shaped larvae hatching rate

Abstract:The optimum measurement for sea cucumber Apostichopus japonicus body weight and coelomic fluid is an essential experiment for the related research. The results show that the body weight is suitable to weigh in 20～30 min after being taken away from culture water (below 20 ℃). The proportion of coelomic fluid to the total body weight was also examined for small, medium and large sized sea cucumbers, at 26.88±1.39%, 25.24±2.91% and 30.98±1.86%, respectively. The safe dose of coelomic fluid extraction was less 1.5% to the total body weight of sea cucumber for ensuring physiological stability and survival of the tested animals. The results illustrated that the total cells of sea cucumber experienced a process of significantly reducing to slowly recovering. The resume process was asynchronous for different kinds of coelomocyte. The results show that the resume period for total cells, granule cells, transparent lymphoid cells and the absorbency of coelomic fluid are 4 d, 6 d, 6 d and 5 d, respectively. The measurement of body weight, the proportion of coelomic fluid to the total body weight, the safe dose and the resume process of coelomic fluid is firstly reported in sea cucumber Apostichopus japonicus. This research could be helpful for further study on physiological, immunological and zoological research in sea cucumber..

Abstract:16S rRNA and COI fragments from two mitochondrial genes of Portunus trituberculatus sampled from three sites (Hokkaido of Japan, the east coast of South Korea and Huichang, Jimo of China) were amplified and sequenced. The variable sites, nucleotide diversity, average number of nucleotide differences and nucleotide diversity index were calculated, and the sequence differences and genetic diversity levels among three different populations were compared. The results show that levels of genetic diversity of Chinese wild population are low. The molecular phylogenetic tree of 16S rRNA was different with that of COI gene, which was constructed using NJ method with software MEGA 4.0. Their main difference was in the classification relationship with Charybdis. The phylogenetic tree of 16S rRNA gene sequences show that Portunidae are clustered into two branches: different haplotypes of P. trituberculatus clustered first, then with P. pelagicus and P. sayi; the three kinds of Scylla clustered first, then with Charybdis japonica. In this study, 19 haplotypes were obtained, and Huichang, Hokkaido and east coast of South Korea populations possessed shared haplotypes, indicating that these three wild P. trituberculatus populations had similar genetic background. These findings are expected to provide molecular biological basis for the protection and utilization of P. trituberculatus resources in China..

Abstract:The fertilization experiment of Tegillarca granosa (♀)× Scapharca subcrenata (♂) was carried out three times at Qingjiang Field Research Station of Zhejiang Mariculture Institute in July, 2009. Consecutive cytological observation of fertilization and embryonic development of Tegillarca granosa (♀)× Scapharca subcrenata (♂) were conducted using both optical microscopy and fluorescence microscopy (Hoechst 33258 staining). The results show that sperm of S. subcrenata can attach to the surface of T. granosa eggs rapidly and the acrosomal reaction can be achieved successfully. Subsequently, the interspecifc sperm activated the meiosis of T. granosa egg and induced the releasing of PB1 and PB2. Then, the male and female pronuclei fused into zygotonuceus and the cleavage began subsequently. The rate of crossing fertilization was approximately 60.2％,and the early embryogenetic development of T. granosa (♀)× S. subcrenata (♂) was slower compared to those of intraspecific control groups, respectively. During the whole development process, the blastula stage was relatively long compared to that of control groups and the embryo was obviously abnormal, which may only reach trochophore stage and died before D shaped larva stage. Trochophore larva of T. granosa (♀)× S. subcrenata (♂) were found to keep excessive cilia growth. In addition, polyspermy and multiple separations were observed in this experiment

Abstract:Portunus trituberculatus；Microsatellite marker；Family；Parentage determination

Abstract:Distribution of giant jellyfish and major zooplankton, the ecological type and dominant species of zooplankton, were analyzed based on the sampling data obtained during the surveys in jellyfish release area of Liaodong Bay, the Bohai Sea, in June and July, 2005~2007. The relationship between zooplankton and giant jellyfish abundance distribution was explored. The results showed that there were three major giant jellyfish, Rhopilema esculentum, Nemopilema nomurai and Cyanea nozakii and three major zooplankton, Acartia bifilosa, Paracalanus parvus and Paracalanus crassirostris in the survey area. Besides, Copepodid and nauplii were other two important zooplankton components. The distribution sites with high catch density of giant jellyfish were usually close to the distribution sites with highly abundant zooplankton.

Abstract:10-week feeding trial was conducted to investigate the effects of four diets containing different combinations of selenium and vitamin E on the growth performance, nonspecific immune responses of Japanese flounder Paralichthys olivaceus and disease resistance when challenged with Vibrio anguillarum. The fish were fed purified diets with supplemented vitamin E at 60 mg/kg (designated e) or 300 mg/kg (designated E) combination with selenium at 0 mg/kg (designated s) or 3 mg/kg (designated S). Each diet was fed to triplicate groups of flounder with initial body weight of 38.5±0.15 g. After the final weighing, 8 fish were randomly taken from each aquarium and injected with V.anguillarum. Mortality was recorded daily for 7 days and nonspecific immune parameters were monitored in the remaining fish. At the end of the experiment, weight gain and phagocytic activity in blood was significantly greater in those fish fed the s/E and S/E diets compared with those fed the s/e and S/E diets. Glutathione peroxidase activity in serum and liver were significantly higher in fish fed the S diets compared with those fed the s diets. High cumulative mortality was found in fish fed the s/e diet than other dietary groups. These results indicate that no synergistic effect of excessive dietary vitamin E and selenium on growth performance, nonspecific immune responses and disease resistance to V.anguillarum in Japanese flounder..

Abstract:A method for the determination of 17 sulfonamides(SAs)in aquatic products by ultra performance liquid chromatography(UPLC)with photodiode array(PDA)detector was developed.The SAs in the samples were first extracted with ethyl acetate，then the ethyl acetate extraction was purified and concentrated by HLB cartridges.The analyses were performed on UPLC with an ACQUITY UPLC BEH C18column(2.1 mm×100 mm,1.7 μm),and the mobile phase was methanol 0.1％ acetic acid aqueous solution.SAs were separated by gradient elution and detected by PDA within 8 min.This method showed a good linearity.For sulfacetamide,sulfadiazine and sulfisomidine,the linearity was in the range of 0.01~2 μg/ml,and for the other SAs,the linearity ranged in 0.01~5 μg/ml.The detection limits of SAs were 2.0~5.0 μg/kg (S/N=3).At the spiked levels of 10 and 40μg/kg,the average recoveries of SAs were 70.4%～91.4％ and 73.3%～92.5％,respectively.The relative standard deviations were less than 10％.Routine tests show that the proposed separation and determination method is rapid,sensitive and efficient and indicate that it is practical for the determination of SAs in aquatic products.

Abstract:To screen out the active parasiticidal components, solvent extractions from Rhizorma paridis were evaluated against ciliates (Zoothamnium sp. and cothurnia) with batch treatment. The bioassay based isolation showed that methanol extract presented 100% antihelmintic efficacy at the concentration of 45 mg/L and the killing active site was proved to be its methanol extract. Meanwhile, the median lethal concentration (LC50) values at 96 h determined from the acute toxicity tests of the killing active site for juvenile giant freshwater shrimp was 272 mg/L. The safety concentration was 111.5 mg/L. The results obtained in this study show that Rhizorma paridis could be a potential plant based medicine for ciliates control.

Abstract:Esterase from Bacillus sp. was immobilized with sodium alginate, gelatin and chitosan. The enzyme, immobilized with 2% chitosan, 1% glutaraldehyde and chitosan microspheres∶enzyme loading=4∶3 (g/ml) exhibited the highest activity recovery (66%). The immobilized esterase with chitosan could be reused for 10 times, and 70% of enzyme activity remained after the 10th batch, which exhibited good operational stability. The immobilized esterase can catalyze α-naphthyl acetate in non-aqueous system, and it displayed high activity in isooctane, n-octane and n-hexane.

Abstract:With the expansion of the sturgeon aquaculture scale in China, the sturgeon industrial upgrading through strengthening of the sturgeon processing technology, development of industry chain, and optimizing of inductrial structure has already became the inexorable trend. Caviar is the leading product of the sturgeon processing industry, so caviar processing on a large scale is an important way in accelerating the sturgeon industrial upgrading. The status of sturgeon caviar industry in the world is reviewed in this paper. Based on the analysis on the latest status of industrialization development of sturgeon caviar processing in China, countermeasures to the main problems of lack of farmed sturgeons for caviar production, slow progress of sturgeon deep processing technology research, underdevelopment of sturgeon caviar brands, and the urgent need of revising the industry standard of sturgeon caviar processing are proposed.