Abstract:The gene fragments of mitochondrial DNA control region of 47 individuals of Sebastiscus marmoratus from eastern Guangdong coastal waters were amplified, sequenced and analyzed.By comparing with 10 corresponding D-loop gene sequences of Sebastidae from GenBank, the taxonomic status of Sebastidae was examined. Fragments (538~544 bp) of control region in mtDNA from 47 individuals of S.marmoratus were determined.A total of 69 variable nucleotide sites and 36 sites with gaps/missing were found and 38 haplotypes were defined. The relative amounts of A, T, C and G bases were 34.5 %，29.0 %，15.8 % and 20.7 %, respectively. The average number of haplotype diversity (Hd) and nucleotide diversity (Pi ) were 0.978 and 0.019 2，respectively. The molecular phylogenetic trees of Sebastidae constructed with H. armatus as outgroup showed that S. marmoratus was basal in the tree and differentiated earlier from their common ancestor. Sebastidae was monophyletic and 11 species were divided into four clades including genus Sebastes, Sebastiscus, Hozukius and Helicolenus, which was consistent with the traditional taxonomic results. In conclusion, mtDNA control region is suitable for the analysis of S.marmoratus not only in population genetic diversity but also in phylogenetic relationship.

Abstract:Lophius litulon is an important benthic fish species which distributes in the northwest Pacific. From May 2008 to February 2009, a total of 663 L. litulon samples at body lengths of 45 ~ 650 mm were collected from the East China Sea and the Yellow Sea. Based on the stomach content analysis, feeding habits and ontogenetic variation of L. litulon were investigated by chi-squared test, one-way ANOVA test, and cluster analysis. More than 120 species of prey were ingested by L. litulon. Osteichthyes and Natantia were the major important prey groups. At species level, the dominant preys were Pseudosciaena polyactis,Champsodon capensis, Apogonichthys lineatus, Acropoma japonicum, and Trichiurus haumela.No significant difference was found for feeding intensity between different regions or different size-groups, but the diet composition was significantly different among three regions in ingestion of Osteichthyes and Natantia, and significant ontogenetic differences among the ingestion of Crangon affinis, Palaemon gravieri, A. japonicum, P. polyactis, and T. haumela. Cluster analysis on ontogenetic variation revealed two major groups, one group consisting of L.litulon at body lengths shorter than 250 mm, the other group at BL longer than 250 mm. The food diversity index, diet composition, and food weight of prey species varied with the increasing of body length. Thus, it can be concluded that ontogenetic diet composition variation follows the “optimal foraging theory”.

Abstract:In May, August and October 2006~2010, thirty integrated surveys of coastal water environment were carried out in Yantai Sishili Bay (YSB) twice per month. Mornitored items included water quality, phytoplankton, surface chlorophyll-a and hydrometeorology. The spacial and temporal distribution of chlorophyll-a and primary productivity in YSB was studied. The results showed that phytoplankton density, chlorophyll-a concentration and primary productivity were generally higher in August; But in 2008，the maximum of phytoplankton density appeared in October, and in 2009 the maximums of both chlorophyll-a concentration and primary productivity appeared in May. In October 2008, the phytoplankton density, the highest during the survey period, reached 51.61×104 cells/L, and significant differences were found among all six sampling sites（P<0.01）; The lowest value (0.02×104 cells/L) appeared in October 2007. From 2006 to 2009 both chlorophyll-a concentration and primary productivity showed a growing trend, but in 2010 they decreased to a similar level as 2008. In the last five years，phytoplankton density, chlorophyll-a concentration and primary productivity were generally higher in east YSB than in the west. Furthermore, a significant positive correlation was found between primary productivity and chlorophyll-a concentration (R=0.921,P<0.01), and a regression equation was developed. It will offer an option to estimate and assess the primary productivity of the coastal waters.

Abstract:The morphological parameters of 30 Hexagrammos otakii individuals ( body length between 22.3 and 36.5cm ) were observed and measured. The structure of their digestive system was observed by anatomic method. The correlation equation of total length and body length was: LB= 0.848 9LT+1.030 9( R2 = 0.978 ). The correlation equation of weight and body length was: W= 0.021 1 L2.992 7B（R2=0.833 7）. The countable characteristics includes dorsal fin rays, anal fin rays, pectoral fin rays, ventral fin rays, caudal fin rays, lateral line, lateral line scales, pyloric caecum, and the first gill arch raker. The parameters were as follows: dorsal 38～43, anal 18～22, pectoral 5～7, ventral 17～18, caudal 13～15; lateral line 5; lateral line scales 80~128; pyloric caecum 27~36; the first gill arch raker 16~19. The digestive tract is consisted of clearly distinguishable oesophagus, stomach and intestine; the intestine twisted two times and the relative length was 0.801±0.01. The morphology of the digestive tract was consistent with the specie's flesh-food habits.

Abstract:A comparison analysis of trace elements in muscle, liver and eggs of wild and cultured Paralichthys olivaceus broodstock was conducted. The results showed that contents of Se and Fe were significantly higher, while Cu was significantly lower in liver of wild fish than cultured fish (P<0.05), and no difference was found for other elements (P>0.05); Contents of both Se and Mn in dorsal muscle of wild fish were statistically higher than those of cultured fish (P<0.05), while no change was found for other elements (P>0.05); Se, Fe and Mn content in eggs of wild fish was about 1.7, 2.0 and 2.8 times higher than those of cultured fish, respectively (P<0.05). Cr content in eggs of cultured fish was 1.78 times higher than those of wild fish, however, no differences were found for Cu, Cd and Pb contents in eggs of wild and cultured P. olivaceus broodstock.

Abstract:A five-valent vaccine was made of Vibrio harveyi, V.anguillarum, Ewardsielatarda, V.scophtalmi and V.alginolyticus which were firstly inactivated by formalin, andthen mixed with astragalan-adjuvant. Turbot Scophthalmus maximus was subjected to vaccines via intraperitoneal injection, and the vaccines were respectively appended with(Group 2) or without astragalan-adjuvant（Group 1). Antibody titers, activities of lysozyme and superoxide dismutase were examined on the 7th, 14th, 21th, 28th, 35th, 42th, and 56th day after firstvaccination. Finally, all the fish were challenged with the five kinds of bacteria (mentioned above) on 56d. The results showed that the activities of lysozyme and superoxide dismutase of both groups reached the peak value on 28d, in which Group 2 were significantly higher than those in Group 1 (P<0.05）. The antibody titers of both groups also reached the highest on 28d, but which were much higher in Group 2 than that in Group 1 (P<0.05）. In Group 2, the antibody titers of five kinds bacteria were 27.25,26.75,28.5,28.75,26.25 respectively, and in Group 1 they were 26, 25.75, 26.75, 27.5, 25. The RPS of Group 2 was 50.0%, 50.0%, 71.4%, 62.5%, 44.4% respectively, and 37.5%, 30.0%, 42.9%, 50.0%, 33.3% in Group 1. The results suggest that the five-valent vaccine can enhance the immunity in turbot Scophthalmus maximus. In addition,the immune enhancement was markedly improved in astragalan-adjuvant group, indicating that astragalan-adjuvant has a positive effect on immunity.

Abstract:In order to reveal the effect of nocardiosis on Pseudosciaena croceas physiological conditions，the process of physiological and biochemical pathology was explored in this article. With the method of enzymology analysis, the change of enzyme activity in several main lesion tissues of P. croceas infected with Nocardia was tested. The results indicated that, in contrast with the control group, the amylase activities had no significant change in heart and spleen, while decreased apparently in kidney, liver and gill(P＜0.01).Lysozyme activities in heart, liver, kidney and gill increased significantly(P＜0.01), whereas declined significantly in spleen (P＜0.01). Alkaline phosphatase activities in heart, spleen, liver and gill increased significantly, while became lower in kidney (P＜0.01). Acidic phosphatase activities were higher in heart, spleen, and lower in kidney and liver,while no significant change in gill was found. Both glutamic-oxalacetic transaminease and glutamic-pyruvic transaminase increased significantly in kidney and decreased apparently in liver(P＜0.01), while none of them showed significant change in heart, spleen and gill(P＞0.05). Malondialdehyde both in kidney and liver decreased and had no significant changes in heart, spleen and gill. Superoxide dismutase activities reduced significantly in liver and gill(P＜0.01), while no significant changes in heart, spleen.The catalase activities reduced significantly in heart and kidney, and increased in spleen(P＜0.01), while no significant changes in liver or gill.Therefore, P. croceas had obvious stress response to Nocardia infection and showed different levels of damage to the organs of sicken fishes.

Abstract:The mitochondrial genomic DNA of Euphausia pacifica was obtained by long PCR amplification, and was then sequenced using shotgun and primer-walking strategies. The mitochondrial genome of E. pacifica is 16,898 bp in length. The largest non-coding region in E. pacifica mitochondrial genome contains one section with tandem repeats (4.7×154 bp). Nad5 gene has the largest number of different loci (319~321), followed by nad4 (284~285) and cox1 (232~233) gene. Therefore, nad5 and nad4 genes can be used as alternative molecular markers to analyze genetic diversity among krills species and populations. Translocation of three tRNAs (tRNALeu(CUN), tRNALeu(UUR) and tRNATrp) is shared by E. pacifica and E. superba mitochondrial genomes when compared with the pancrustacean ground pattern. The duplication of tRNAAsn and translocation of tRNAIle were found in the mitochondrial genome of E. superba when compared with the E. pacifica mitochondrial genome. Gene orders are not identical between E. pacifica and E. superba mitochondrial genomes, indicating that gene order is not conserved among euphausiids mitochondrial genomes.

Abstract:A pathogenic bacterium strain 20100612001 was isolated from the hepatopancreas tissue of diseased white shrimp Litopenaeus vannamei. The LD50 challenge assay was performed and the LD50 value was found to be 1.44×106CFU/ml. The cell morphotype was observed to be bacilli，coccoid rod or pear-like and some cells could be observed intensely stained at two poles. The strain was Gram-negative, sporeless and had a single polar flagellum. It formed transparent or translucent round colonies on 2216E solid medium but green or blue-green colonies on TCBS medium. It showed the highest identity to V. parahaemolyticus or Vibrio natriegenis by using fatty acid analysis and Biolog system. Based on 16S rRNA sequencing, the strain is >98.9% identical to several species in Vibrio genus, and the phylogenetic tree revealed that it is most close to Vibrio parahaemolyticus. Taken together, the pathogenic strain 2010060612001 is identified as V. parahaemolyticus.

Abstract:The salinity tolerance of “Huangxuan No.1” Portunus trituberculatus was investigated in controlled temporary water by including sudden and gradual changes in salinity. The results showed that the LD50 values of crab larvae at Stage Ⅱ for 24h,48h,72h tolerance to low-salinity were 21.655, 22.109, 23.184，respectively, and for tolerance to high-salinity were 50.711, 50.061, 49.612 respectively. The LD50 values of 80-day crabs for 24, 48, 72h tolerance to low-salinity were 5.13, 7.49, 8.56，respectively, and for tolerance to high-salinity were 54.49, 52.74, 52.21 respectively. Crab larvae at Stage Ⅱ can survive in 6.7 low-salinity water by gradual salinity decrease,and can survive in 47.7 high-salinity water by gradual salinity increase.While 80-day crabs can survive in 5.7 low-salinity water by gradual salinity decrease,and can survive in 47.7 high-salinity water by gradual salinity increase. Therefore,“Huangxuan No.1” P. trituberculatus has a strong salinity tolerability.

Abstract:Bio-floc technology was applied in closed culture system of Litopenaeus vannamei. The optimum carbon source and its addition for bio-floc culture was screened, and the optimum carbon addition and stocking density of L. vannamei was determined. Results showed that in closed culture system of L. vannamei at the stocking density of 150 and 300 PL/m2, sucrose was added in the quantity of 77% feed (containing 42% protein) everyday, bio-floc formed on the 4th day post stocking. During the 84 d culture period, the ammonia-N and nitrite-N concentrations kept at very low levels, and the shrimp survival rate was above 80%, which means better harvest of L. vannamei in bio-floc culture system.

Abstract:Astakine (AST) is a cytokine that promotes cell differentiation and proliferation of hemocytes in the hematopoietic tissue, which plays important roles in crustacean immune responses. Evidence from our previous studies showed that WSSV may compete with AST for a common receptor. By rapid amplification of cDNA ends (RACE) PCR, the full-length gene of Litopenaeus vannamei astakine gene (lvast) was obtained, which consisted of 1,846 bp with a 372 bp ORF, encoding 124 amino acids with the predicted molecular mass of 13.3kDa. The deduced protein contained a prokineticin (PK) domain. Homological comparison among LvAST and other species showed that the homogeneities were higher in comparison with those of Penaeus monodon and Pacifastacus leniusculus, and lower in comparison with those of vertebrates.The recombinant protein of LvAST was successfully expressed with the constructed recombined vector pBAD/g IIIA-lvast containing the lvast ORF. The results laid a theoretical basis for further functional studies of astakine.

Abstract:Aquaculture of abalone was limited in northern China by the lack of fresh food in summer. For a new feeding method, the Pacific abalone Haliotis discus hannai Ino at shell length of 65mm were treated with periodic starvation, i.e. feeding for 2d, then starving for 1~4d, with continuous feeding treatment as control. Results of growth, feeding, defecation and granulocyte suggested that there was no significant difference in growth when starvation was less than 2d (P＞0.05), while body weight dropped in other groups(more than 3d starvation). Food intake rate increased as starvation lasted, but there was no significant difference between each group (P＞0.05). Fecal production rate and organic matter in feces decreased with prolonged starvation, and it was significantly different during each treatment stage (P＜0.05); Ratio of granulocyte showed no difference in treatments of starvation less than 2d, and significantly higher for group of starvation less than 2d than groups of f2s3 and f2s4 (P＜0.05).

Abstract:Survival, growth and feeding rate of sea cucumber Apostichopus japonicus were studied in two different salinity change modes (DG: salinity Decreased Gradually; DS: salinity Decreased Sharply). In each mode, sea cucumbers were kept in seawater at 4 different salinities (31-A, 26-B, 22-C, 18-D) for 30d. The results showed that there were only a few individuals died in group B. There was significant difference between group C and D in DG and DS modes (P<0.05). The lowest survival rate was in group DS-D as 45.67%. There was significantly different FR and FCR between treatments under different low salinity stress (P<0.05). However, no significantly different FR and FCR appeared between different salinity change modes (DG and DS) during the experimental period (P>0.05). In group B, there was significant difference on growth of sea cucumber between two salinity change modes in the first 10d after the salinity decreased (P<0.05), but no significant difference in the next 20d (P>0.05). During the experiment, there were negative growth in group C and D, being significantly different from the other groups (P<0.05).

Abstract:A multiplex polymerase chain reaction was developed for the simultaneous detection of five major foodborne pathogens, Vibrio parahaemolyticus,Staphylococcus aureus, Listeria monocytogenes,Salmonella enteritidi, and Shigella flexneri. Five pairs of specific primers were designed according to the thermolabile hemolysin gene of V. parahaemolyticus, the heat stable nuclease gene of S. aureus，the Listeriolysin O gene of L. monocytogenes, the invasion protein A gene of S. enteritidis, the invasion plasmid antigen H gene of S. flexneri. The reaction conditions were optimized, and the specificity and sensitivity of this method were tested.The detecting results of 48 samples also proved its reliability and practicability.

Abstract:The lolB gene has been shown to code an outermembrane lipoprotein, and serve as a reliable molecular marker for the detection of all V.cholerae serogroup and biotypes. A pair of specific primers based on lolB gene of V.cholerae was designed, and a real-time PCR using SYBR Green I for V.cholerae detection was established. A 519bp gene fragment was amplified from chromosomal DNA of V.cholerae, and no positive reaction was detected in 3 other pathogenic Vibrio species using conventional PCR. The melting curve analysis of SYBR GreenⅠreal-time PCR showed one specific peak with melting temperature（Tm）of 86.5~87 ℃, and no primer-dimers peak present. The results indicated that the PCR primers have good specificity. Amplification curves of SYBR GreenⅠreal-time PCR revealed the geometric phase and plateau phase of PCR. Analysis of standard curves revealed excellent correlation between the quantity of bacteria (2.59×108 to 2.59×100) and PCR threshold cycle (Ct),with the correlation rate of 0.993. The results indicated that the SYBR GreenⅠreal-time PCR could be used as an effective assay for quantification of pathogenic V.cholerae. The assay could be completed within 4~5 h from extraction of nucleic acids to analysis of results. The SYBR GreenⅠreal-time PCR was a simple, rapid, specific and sensitive method for the diagnosis of aquatic animal diseases and investigation of epidemics caused by V.cholerae.

Abstract:The panel test on fouling organisms on five materials (epoxy board, HDPE, FRP, carbon steel and cement block) was carried out in offshore of Yantai Economic and Technological Development Zone(ETDZ) from May to July 2010.A total of 16 fouling organism species were recorded, among which Tubularia marina, Cryptosula pallasiana, Mytilus edulis and Hiatella orientalis were the dominant species. Among four pipe materials in seabed, cover area of the fouling organism on cement bricks was the largest, followed by FRP, HDPE, and carbon steel. The attachment of fouling organisms reached the peak from early July to mid-July. FRP or HDPE is recommended for water supply and drain pipe.