Abstract:A wide variety of compounds were used to anaesthetize fish in fisheries research, fisheries management, aquaculture and fish health. Among of them, tricaine mesilate ( MS-222) is one of the anaesthetics widely used in the world for it's safety and high effectiveness. In China, little published information was found on the use of MS 222 in marine fish. In this study, immersion anaesthesia was utilized to investigate the recovery time, recovery rate and survival rate in 24 h of flounder Paralichthys olivaceus after being anaesthetized with different concentrations of MS 222 at the temperature of 13-14 ℃; and the effect of long time immersion anaesthesia in 60 mg/L MS-222 on the recovery rate and survival rate after being transferred from water into air were studied. The results showed that the lowest effective concentration to anaesthetize the flounder was at 50 mg/L. The time to achieve anaesthesia was shortened from 288 s to 50 s and the time to recovery was extended from 94 s to 106 s when the concentration of MS-222 was increased from 50 mg/L to 200 mg/L. Three concentrations of MS-222 including 60 mg/L, 100 mg/L and 200 mg/L were used for long time immersion anaesthesia respectively to evaluate the recovery time, recovery rate and survival rate in 24 h, the results showed that the recovery time in the three groups were prolonged along with the increasing of immersion time, but the longest immersion time to achieve 100 percent recovery rate were 64 min, 32 min and 32 min respectively. Also the time for enduring air exposure was tested after the flounder was exposed in 60 mg/L MS 222, and it was found that their ability to endure air exposure was better than the control. Recovery time showed great difference between individuals and the survival rate in 24h after anaesthesia was 100% once the flounder was recovered. These results indicated that the ideal concentration of MS-222 is 60 mg/L, the time to keep the flounder in MS 222 and the exposed time to air is less than 64 s and 60 min respectively at this concentration.

Abstract:The effects of temperature and pH on protease and amylase activities in skewband grunt Hapalogenys nitens was studied.Results showed that the pH values of the stomach, hepatopancreas and intestine were 4.9～5.4,5.8～6.2；and 6.5～6.9 respectively. Protease activity rose with the increase of water temperature with a peak at 40 ℃,and then decreased. The order of protease activity was as follows: stomach＞foregut＞pyloric caecum＞hindgut＞hepatopancreas. The optimum temperature for amylase in H. nitens was 35 ℃ within the range of 15～50 ℃; the order of amylase activity was as follows: hepatopancreas＞pyloric caecum＞foregut＞hindgut＞stomach.The optimum pH for protease activity in stomach was 2.8 ℃ within the range of 2.2～7.6, while the optimum pH for protease activity in other digestive organs was 7.2 ℃ with in the range of 4.8～8.0. Under the optimum pH conditions, the order of protease activity in different organs was as follows: foregut＞pyloric caecum＞hindgut＞hepatopancreas. The optimum pH for protease activity in stomach was 6.0 in the range of 4.8～8.0. The optimum pH for amylase activity in stomach was 6.0 while it was 6.8 in intestine, hepatopancreas and pyloric caecum. The order of amylase activity was as follows: hepatopancreas＞pyloric caecum＞ foregut＞hindgut＞stomach.Under the optimum temperature and pH conditions, the order of protease activity was as follows: stomach＞foregut＞pyloric caecum＞hindgut＞hepatopancreas, while that of amylase activity was: hepatopancreas＞pyloric caecum＞foregut＞hindgut＞stomach.

Abstract:Seasonal species composition of crustacean was analyzed based on the data from the fishery resource monitoring investigation carried out in spring (April), summer (June), autumn (September) and winter (December) of 2004 in the East China Sea and the Yellow Sea. A total of 33 species of shrimp, 12 species of crab and 3 species of stomatopod were collected. The dominant species in the East China Sea and the Yellow Sea was identified as: Solenocera melantho, Parapenaeus fissuroides, Trachypenaeus curvirostris, Metapenaeopsis longirostris, Solenocera koelbeli, Crangon hakodatei, Plesionika izumiae, Solenocera alticarinata, Charybdis bimaculata, Portunus trituberculatus and Oratosquilla oratoria. Among theses, S. melantho, P. fissuroides and T. curvirostris were the main dominant species in the survey area. The biodiversity 〖JP2〗of crustacean in the East China Sea and the Yellow Sea was indicated by 3 indices: Margalef index (D), Shannon Wienner index (H') and Pielou's evenness index (J'). D and H' trended to decrease from the northern to the southern area in spring, increased from the north to the south in summer and autumn, and showed high value in the north of the East China Sea (ECSN) and low value in the south Yellow Sea (YSS) and the middle of the East China Sea (ECSM) in spring. J' showed little difference among seasons in all areas. The community of the ECSM showed a relative stabilization, while the community characteristics of ECSN varied with season.

Abstract:The extracellulose products (ECP) of Aeromonas salmonicida masoucida was extracted by using cellophane plate technique. It was proved to be lethal to swordtail fish (Xiphophorus helleri), and the LD50 does was 4.72 g protein/g. The ECP was analyzed by SDS PAGE and thirteen proteins were observed. Western blot with rat's antiserum against ECP showed that seven proteins among those thirteen proteins, whose molecular weight were 88, 70, 42, 39, 36, 22 and 15 kDa,respectively, were considered to have immunogenicity and could activate the immunoreaction of the hosts to secrete antibody.

Abstract:A series of experiments have been conducted to study the effects of salinity and temperature on embryonic development and starvation endurance of newly hatched larvae of rock bream Oplegnathus fasciatus (T.& S.). Experiments were carried out in Qingdao Zhonghai Fishery Co., Ltd. from 2004~2007. Eggs were collected immediately after spawning and transferred into tanks for incubation. Salinity study was carried out in 2005, with treatmentsat salinities of 40，35，30，25，20，15，10 and 5.The results showed that the suitable salinities for embryonic hatching ranged from 20 to 35 with the most suitable range between 25～30, when hatching rates were all above 88%. Salinities above 40 or below 20 led to low hatching rate and prolonged hatching time. Temperature study was done in 2004,2005 and 2007, respectively. Taking into consideration the hatching rate,hatching time and deformity rate of the newly hatched larvae, it is deduced that the suitable temperatures for hatching ranged from 21～27℃ with hatching rates above 77%;most suitable temperatures were 23～25℃ with high hatching rates over 89% and low deformity rate of newly hatched larvae. Hatching time was negatively correlated with temperature while embryonic development speed was positively correlated with temperature. The threshold temperature for embryo development was calculated as 10.06℃, the effective accumulated temperature was 4053℃•h. In addition, Q10 value was calculated, and was found to accord with the experimental result. Newly hatched larvae were starved to death at 8 DAH with half death time appeared at 6 DAH;starvation had regative effects on larval growth.

Abstract:Studies on the toxic effects of suspended substances (SS) on the embryos and newly hatched larvae of Cynoglossus semilaevis  Günther were conducted in laboratory in October 2007. The results showed that the effect of SS on hatching time was not evident. When the SS concentration was less than 200 mg/L, there was no significantly difference in hatching rate (P＞0.05)between the treatment and control.When the SS concentration was more than 200 mg/L, there were significant difference between the treatment and control in embryos hatching rate and deformity rate of newly hatched larvae (P＜0.01). The Incipient LC50（95% confidence limits）of SS to embryo was 1 974.8 mg/L (1 375.4-2 835.5 mg/L), and the safety concentration(SC) was 197.5 mg/L.The 48 h LC50 and 96 h LC50（95% confidence limits）of SS to newly hatched larvae were 226.9 mg/L (204.6-251.7 mg/L) and 202.9 mg/L (182.3-225.9 mg/L), respectively,and the SC was 20.29 mg/L.

Abstract:The biological characteristics and phylogenetic analysis of bacteria isolated from Epinephelus lanceolatus L. were studied, including morphological characteristics, physiological and biochemical characteristics. In addition, the 16S rRNA gene were partially sequenced and compared with sequences registered in databases; and molecular phylogenetic trees were constructed. The results showed that the two pure strains （HQ061227 1、HQ061227 2）belonged to P. damselae subsp. damselae (Love et al. 1982; Smith et al. 1991) of Photobacterium (Beijerinck〓1889 ), the sequenced 16S rRNA gene of strain HQ061227 1（accession No. EF635307) is 1 469bp in length, and exhibited high similarity (99%) with the 16S rRNA gene of P. damselae from GenBank database. Antibiotic sensitivity using 37 antimicrobial agents showed that the isolates were sensitive to 32 agents including cefazolin, resistant to 4 agents including penicillin G, and slightly sensitive to ampicillin.

Abstract:Horizontal starch gel electrophoresis was used to investigate the genetic structure 〖JP2〗and differentiation of the three〖JP〗 populations (Qingdao, Zhoushan and Xiamen) of Nibea albiflora. Eleven putative loci were revealed by eight enzymes (G3PDH, IDHP, LDH, MDH, PGDH, PGM, SDH and SOD). The percentage of polymorphic loci for each population was 02. The mean observed and expected heterozygosity ranged from 0010 1 to 0037 9 and from 0023 4 to 0039 9, respectively. The average number of efficient allele for each locus varied from 1.029 0 to 1.052 8. The genetic distances among populations were from 0.000 4 to 0.000 8, and the genetic similarities were from 0.999 2 to 0.999 6. The results showed that there was no significant genetic differentiation among the three populations of N. albiflora

Abstract:The diseased Japanese flounders Paralichthys olivaceus were obtained from a culture farm near Qingdao. The lymphocystis nodules were cut down and washed with TNE buffer (50mmol/L Tris, 100mmol/L, 1mmol/L EDTA, pH 7.4), about 10 gof nodules were homogenized and suspended in 90 ml TNE buffer at 4℃. The suspension was subjected to three rapid freeze/thaw cycles. Following ultrasonication, the virus cell debris suspension was centrifuged at 650×g for 20 min at 4 ℃. Then the supernatant was centrifuged again at 1 800×g for 20 min at 4 ℃, pooled and stored overnight at 4 ℃, while the pellet was discarded. Virus particles were pelleted from the pooled supernatant by ultracentrifugation at 78 500×g for 120 min at 4 ℃ and resuspended in TNE. The suspension was equally divided and loaded onto pre formed gradients made up of 33%, 40%, 47%, 52%, 57% and 62% (w/w) sucrose, and ultracentrifuged at 78 500×g for 120 min at 4 ℃. Two prominent bands were obtained in sucrose gradients after ultracentrifugation. A diffuse band (B1) was located at about 37%~40% sucrose concentration, while a distinct band (B2) was located at about 47%-50% sucrose concentration. B1 contained few viral particles together with amorphous material, membranous structure and other cell debris when assessed by electron microscopy, while B2 from sucrose gradients contained viral particles and was apparently free of other material. In electron micrographs, the virions were intact, round polygon in shape, about 210-250 nm in diameter, some of the viral particles appeared with a more or less hexagonal profile.  In addition, for learning immunological characterization of LCDV purifitied from different zones，the rabbit antiserum was raised against the LCDV purified from Weihai region. Serum homology of LCDV isolates from Qin Huangdao, Qingdao and Weihai were checked by western blotting. Western blotting profile showed three main antigenic proteins with molecular weights of 125, 66 and 55kDa reacted with the antiserum. This experiment showed that the virus from different zones had the same linear epitopes.

Abstract:A pair of PCR primers, with EcoR I and Not I restricted enzyme sites in the 5， and 3， end respectively, was designed according to the major capsid protein (MCP) gene of turbot viral reddish body iridovirus (TRBIV). After PCR amplification, a recombinant expression vector including α factor peptide and 6×His tag was constructed by inserting TRBIV MCP gene into GAP promoter downstream of pGAPZαA directly. The constructed vector pGAPZαA MCP was linearized by Avr Ⅱ, and then transformed and intergrated into the host Pichia pastoris X 33 genome by electroporation. The recombinant yeasts with high level secreted expression of MCP were screened by ELISA Dot. The expressed proteins were identified by SDS PAGE and Western blot. The results revealed that a recombinant protein with the molecular weight of approximately 51kDa was secreted into the supernatant from the recombinant yeast cells successfully. The expressional amount of TRBIV MCP could reach to 60.2 μg/ml in the supernatant secreted from recombinant yeast after being fermented at 28 °C for 72 h. 

Abstract:The study is based on the bottom trawl catch data in Jiaozhou Bay from 2005 to 2006. The Sepiella maindroni abundance in the waters of Jiaozhou Bay was assessed by using resource density area method. The ecological characteristics and seasonal distribution were surveyed； the catch composition of bottom trawl were compared；the species composition, seasonal variations and distribution were analyzed. The results showed obvious changes in Jiaozhou Bay Sepiella maindroni resources in different season；the resources in summer and autumn respectively reached 26.7 t, 21.5t (2006) and was significantly higher than that of the other seasons.The proportion of weight in summer and winter were 23.4% and 278%,respectively,and was obviously higher than that of the other seasons in 2006. In recent years，the cephalopod resources in which the Sepiella maindroni was a main catch species maintained almost stable and the average catch was about 14t, close to the level of mid 1990s. 

Abstract:With α naphthyl acetate and tributyrin as substrate，a precise and rapid screening method was developed．One stable strain EB 1，which was isolated from related samples，showed the highest esterase producing ability among the 281 strains．Through study on its morphological，physiological and biochemical characteristics as well as 16S rDNA sequence homology comparison，strain-EB-1 was identified as Bacillus subtilis var.niger，which was almost the closest relative to the standard strain with 9979% similarity in sequence under the phylogenetic tree．The 16S rDNA sequence of strain EB 1 has been deposited in GenBank，registration number was EU016526．

Abstract:At temperature 24±0.5℃, the filtration rates of scallop Chlamys farreri spats (1.177～2.017 mm in shell length) were determined at different salinities (20,25,30,35) when microalgae density was 3.0×10⁴ cell/ml and different density of microalgae (Dicrateria zhanjiangensis) (3.0×10⁴ cell/ml, 4.5×10⁴cell/ml, 6.0×10⁴ cell/ml) when salinity was 30, using static water method. The relationship between the filtration rate (FRS) and the salinity (S) can be expressed byFRS＝-30.893S²+1 691.5S-196.10 (r=0.847) (calculated by whole dry weight), FRS＝-0.022S2 1.223 6S-14.522 ( r= 0.928, calculated by individual quantity) . We estimated that at the salinity of 27.8，the filtration rate reached the maximum value, 3.54L/g•dw•h（2.49×10^-4 L/ h•ind）. The filtration rate of the scallop spats increased with the salinity when the salinity was lower than 27.8 and decreased when it was more than 27.8. Feeding rate had significant influence on the filtration rates of the scallop (P< 0.05). When fed with D.zhanjiang ensis at in the concentrations of 3.0×104cell/ml, 4.5×104cell/ml, 6.0×104cell/ml, the filtration rate of the scallop spats increased and then decreased in a parabolic trend.It was estimated that the highest filtration rate(0.43×10^-3L/ h•ind)could be reached when the density was 4.7×10⁴ cell/ml.The relationship between the filtration rate (FRQ) and the microalgae density (Q) was FRQ=-0.069 3Q² 0.648 4Q-1.083 5(r= 0.722)。

Abstract:The eggs of Chlamys farreri were activated by ultraviolet irradiated heterogenous sperms of Pacific oyster Crassostrea gigas and the pro embryos were induced by inhibiting extrusion of the second polar body from fertilized eggs with 6 DMAP. Those eggs were treated with different parameters of 6 DMAP which were designed in the method of orthogonal experiment design L9（34）. It was found that the occasion of starting time, the length of the treatment time and the concentration of 6-DMAP were important factors to affect the derivational rate of diploids. The statistic data of the trochophores' chromosomes in the nine groups was analysed with the software SPSS115. Results showed that all the three factors had prominent effect on the derivational rate of diploids; the relationship between the rate of malformed embryos and the rate of diploids was negative correlation and the equation was y=46.632-0.891x (r=-0.813，P＜0.01）. Based on the relationship between derivational rate of diploids and the rate of malformed embryos, in the stated range, it was confirmed that the optimal inducing condition of 6-DMAP for gynogenesis in the C. farreri was that the eggs were to be treated with 6-DMAP at the concentration of 60μg per ml for 25 minutes when of the fertilized eggs expelling first polar body 20% to 25%. In this way, 29.5±5.36% diploidinduction could be got. In this study, the states of poly ploidy yield in the treatment groups were also discussed.

Abstract:A self designed culture system was used for the study of intensive indoor cultivation of Calanus sinicus. Water flow and temperature of the circulating system was automatically controlled. During the course of 61 d laboratory experiment, the number of adult Calanus sinicus increased from 2.3 ind/L to19.7 ind/L, and the egg production increased from14.7 eggs/L to 85.7eggs/L, respectively. Major water quality parameters, such as DO, NH4-N and CODcr were kept at an acceptable range. The results indicated that the establishment of this culture system might lay a foundation for intensive indoor cultivation of Calanus sinicus.

Abstract:Based on four surveys from August 2003 to May 2005 in Sanggou Bay，Shandong Province, the ecological characteristics of species composition, biomass，community structure of phytoplankton were studied. The results showed that there were 72 species belonging to 32 genera of phytoplankton in the surveyed areas. Among them, 60 species of 29 genera were Bacillariophyta,which accounted for 83.3% of all species, and 12 species of 3 genera were Pyrophyta, which accounted for 16.7% of all species. The mean values of the phytoplankton biomass were 5.0×10⁴～3 416.0×10⁴ cell/m³；the maximum appeared in August 2003 and the minimum appeared in May 2005.The analyzing results diversity index，abundance index，simplicity index and evenness index showed that the characteristics of community structure varied with different seasons.The species number,biodiversity of phytoplankton in autumn were obviously greater than that in spring.The comprehensive assessing result suggests that the quantity distribution of species in Sanggou Bay was not even,and the biodiversity of phytoplankton was in moderate grade.

Abstract:In order to understand antioxidant enzymes activities in response to temperature increment and the active oxygen free radical clearing function of antioxidant enzymes in antarctic ice mircoralgae Chlorophyceae L4, the free radical generation rate, the malonaldehyde (MDA) content changes and the corresponding antioxidant enzymes activities changes in Chlorophyceae L4 at different temperatures were determined. The results showed that the free radical generation rate and MDA content in Chlorophyceae L4 increased rapidly in early time (24 to 72 hours) of temperature increment, then decreased rapidly, and could return to tolerable leve1 at the end of time. Under the stress of high temperature, the superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) activities in Chlorophyceae L4 slowed down sharply in early time, then increased obviously, which were much higher than those in the control，and their activities became higher at higher temperatures. In a word, the antioxidant enzymes system plays an important role in the adaptation of antarctic ice microalgae to high temperature environment.

Abstract:In order to study the distribution characteristic of temperature, salinity, pH, dissolved oxygen, DIN and PO3-4-P and evaluate the nutrient level and organic pollution situation in Laizhou Bay and its adjacent waters, samples were collected and analyzed in August 2007. the results showed that the concentration of inorganic nitrogen was relatively high in both surface and bottom of west and southwest waters of Laizhou Bay, which showed an decreasing trend from the west and southwest to the northeast. The PO3-4-P in surface waters was well distributed, a small high value area was found in the middle waters of Laizhou Bay, the concentration decreased from the center toward the periphery gradually. Concentrations of PO3-4-P in bottom waters were different from those in surface waters correspondingly, but the scope of high value area was greater and isoline was more intensive. According to the correlation analysis between nutrient and salinity, there was significant negative correlation between DIN and salinity, indicating that the riverine input was the main sowrce of DIN. Since there was not significant correlation betweenPO3-4-P and salinity, riverine input was not the main source of PO3-4-P； the concentration of PO3-4-P was maintained mainly by the decomposition and mineralization of organic materials. In addition, mass reproduction of phytoplankton was another important influencing factor for PO3-4-P. According to water environment evaluation result, the value of N/P ratio was higher than the Redfield ratio of 16,and phosphate was the limiting factor for the growth of phytoplankton. Based on the evaluation mode of eutrophication and organic pollution index, The nutrient level of Laizhon By was medium phosphorus limiting potential eutrophication, and the organic pollution degree remained at the third level in summer 2007, which indicated that the water area had been polluted by organic materials slightly.

Abstract:In China the resource of the scallop mantle which is rich in proteins is so abundant that there is much to do to utilize them adequately. Combinations of any two of neutral protease (AS1.398), trypsin and compound protease were used to hydrolyze the mantle of Chlamys farreri. The free amino acid nitrogen (AAN) in enzymolysis liquid was chosen as indictor of enzymolysis degree. By orthogonal experiments, effects of enzyme amount, reaction time, temperature and the type of proteases on the hydrolysis of the mantle were investigated. The best hydrolysis condition was optimized and then used to prepare amino acid nutrition powder by spray drying.The analysis results showed that in this product there was 82.89 % crude protein, 1.55% crude fat and 6.82% total sugar. Furthermore, it contained all kinds of amino acids，and the content of total amino acids and free amino acids in 100g crude protein was 79.06g and 51.53g.

Abstract:he effects of feeding for 50d on a diet containing different levels of salt (NaCl) were evaluated in low salinity (1.5 ) and sea water (30 ) for L.vanamei. In low salinity water , the addition of salt to the diet resulted in a significant improvement in the SGR, FCR, survival rate and activity of alkaline phosphatase for L.vanamei. In low salinity water , the addition of salt to the diet resulted in an increase in protein and decrease in moisture and lipid of shrimp body. Moreover,in low salinity water , the addition of salt to the diet resulted in an increase in total free amino acids of shrimp body and significant improvement in flavor characters, especially for the addition of 2.0% salt group. In seawater, the addition of salt to the diet resulted in a significant decrease in the SGR, FCR and survival rate and reduced in activity of alkaline phosphatase for L.vanamei.

Abstract:Comparisons between tensile mechanical properties of high strength polyethylene braided netting twine (HSPE braided netting twine) and common polyethylene braided netting twine (PE braided netting twine) for fishing have been made by applying the SC 5005 fisheries trade standard and SC 110 fisheries trade standard.The results revealed that under the same condition(the pitch of twist of twine was 27 mm, the structure of strand was 3×16,the twine core was a parallel thread which was made from 7 monofilaments without twining), the HSPE braided netting twine（whose specification was HSPE－37tex×3×16＋7）was 10.0% and 9.1% higher than common polyethylene braided netting twine (PE braided netting twine PE－37tex×3×16＋7) in breaking strength and overhand knot strength, respectively, while in percentage of breaking elongation, the former was 11.3% smaller than the latter Under the same condition(the pitch of twist of twine was 30 mm, the structure of strand was 4×16,the twine core was a parallel thread which was made from 18 monofilaments without twining), the HSPE braided netting twine（HSPE－37tex×4×16＋18）was 8.1% and 6.1% higher than common polyethylene braided netting twine (PE braided netting twine, PE－37tex×4×16＋18）) in breaking strength and overhand knot strength, respectively, while in percentage of breaking elongation, the former was 13.3% smaller than the latter. Under the prerequisite of strength of braided netting twine, the HSPE braided netting twine, if used in fishing gear or fish cage, is more feasible than the same structure PE braided netting twine.

Abstract:Anatomical and SEM observation of the inflorescence showed that eelgrass was a monoecious plant, the male and female flowers were alternately arranged on the inflorescence. Each anther had one chamber, and the pollen of eelgrass was filamentous, with no exine sculpturing, inaperturate. The carpel contained one ovule and two stigmas, there were filaments on the epidermal cells in front of the carpel. All the characters showed that eelgrass had already adapted to submarine life.

Abstract:Sponge is one of the most primitive multicellular animals in the world, which distributes widely on earth, from tidal zone to deep sea bed, even can be seen in fresh water. This article outlines the biological characteristics of sponge， including external features, internal structure, life habits and so on. The article also introduces the prospect of using the high efficiency pump function of sponge in purifying aquaculture water and as parts of the symbiotic system to prevent aquaculture disease.