Abstract:Epinephelus coioides GH cDNA was cloned from pituitary cDNA library by random sequencing. The sequence obtained spanned 955bp, w ith an open reading frame encoding a protein of 204 amino acids, which is composed of a putative signal peptide of 17 residues and a mature polypeptide of 187 amino acids. The cDNA f ragment encoding the mature polypeptide of GH was PCR amplif ied and subcloned to expression vector pET- 15b ( Novagen) , and expressed in E. coli BL21 ( DE3) as fusion polypept ide containing a His6 at the N - terminus. The addit ion of 0. 4 mmol..L - 1 IPTG induced expression of a protein band with molecular weight of about 24 kDa. The expressed protein accumulated as inclusion bodies, which were solubilized in 6 mol..L- 1 guanidine HCl, and further purif ied and renatured on Ni2 - NTA resin. The purif ied Epinephelus coioides GH fusion polypeptide migrated as a single band of 24 kDa on SDS- PAGE and exhibited GH immunoreactivity in sea bream GH RIA system.

Abstract:The sperm motilities were determined by using phase contrast microscope and analytical system of computer. The result s show the sperm activity rate signif icant ly increases( P < 0. 01) w ith the concentration of NaCl when that of NaCl ranges from 0 to 0. 3% in Monopterus albus, but the sperm activity rate, linear velocity of the spermatozoal mot ion and the frequency of flagellar beat of spermatozoa signif icantly reduce ( P < 0. 05) when the concentration of NaCl exceeds 0. 7% . The micropylar region of the mature egg is like a funnel cave in M. albus under scanning electron microscopy. A micropylar canal is located at the central section of the bottom of the cave, about 4. 22 .. 0. 66 ..m in diameter. The penetration of sperm into the eggs is slow and the process of the fertilization is long. The sperm originally arrives at the surface of the chorion of M. albus 30s after insemination. The micropylar canal is blocked by the fertilized plug within 5 minutes after insemination.

Abstract:In this study, the random amplified polymorphic DNA( RAPD) technique was used to analyze the genetic variation among the allotetraploid hybrid of red crucian carp ( Carassius auratus red var. , .. ) .. common carp( Cyprinus carpio L. .. ) , red crucian carp and common carp. One hundred 10..nucleotide..long random primers were used in the RAPD analysis. Of these primers, 60 primers produced well..amplif ied and reproducible band patterns. There were 6 to 20 bands ranging in size from about 200 to 3 500 bp in the amplified products of a single primer. After excluding bands that were not clearly identifiable, a total of 1 554 distinguishable fragments were compiled for phylogenetic relationship analysis. The genetic similarity index of the amplif ied DNA bands between the allotetraploid hybrid and their original maternal red crucian carp was 69. 41 % , and the genet ic similarity index of the amplified DNA bands between allotetraploid hybrid and their original paternal common carp was 64. 47 % . The similarity index of the amplif ied DNA bands between red crucian carp and common carp was 42. 18 % . Moreover , 4 specif ic bands were found only in the allotetraploid hybrid, the non..parentage bands ratio of the allotetraploid hybrid was 0. 72 % . The results not only provided the specific molecular markers of the allotetraploid hybrid , but also demonstrated that the allotetraploid hybrid inherited a little more genetic materials f rom their original maternal red crucian carp than their original paternal common carp.

Abstract:The coding sequence of green f luorescent protein gene was cloned under the control of carp ..-actin gene promoter. A fish expression of green fluorescent protein gene recombinant was constructed. The recombinant gene was transferred into fertilized zygotes of goldfish ( Carassius auratus) via micro injection after Pvu .. linearing . The green f luorescence was visualized within 48 h following transformation. The transformation and expression of green fluorescent protein was demonstrated by PCR and Southern blotting. In Southern hybridization analysis, transformations got positive signals. The results indicated that the green fluorescent protein gene had integrated into goldfish genome.

Abstract:The trypsin-inhibitory activity of the seven cultured f ish species, Ctenopharyngodon idellus, Carassius auratus, Hypophthalmichthys molitrix , Clarias fuscus, Aristichthys nobilis, Micropterus salmonoides and Monopterus albus were investigated. Individual f ish were bled and the trypsin inhibitory activity analysed using an arginine-aniline dye ester as clorimetric substrate. Results indicated the total trypsin inhibitory decreased from Ctenopharyngodon idellus through Carassius auratus , Monopterus albus, Aristichthys nobilis , Hypophthalmichthys molotrix, Clarias fuscus and Micropterus salmonoides with the lowest level. Analysis of the A2-macroglobulin actvity indicated a ranking w ith decreasing concentration of M. albus, C. idellus, C . fuscus , M. salmonoides, H . molotrix , A. nobilis and C. auratus. The results showed that M. albus had the highest A2-macroglobulin actvity, whereas C. idellus possess a higher level of trypsin-inhibitory activity than the other f ive species.

Abstract:The 13 tissues ( liver, kidney, muscle, bone, skin, heart , lung, intestine, blubber, spleen, pancreas, fore stomach and main stomach) of 22 Dall. s porpoise ( Phocoenoides dalli ) ( 11 ` and 11 a ) off Sanriku coast of northern Japan were measured for iron ( Fe) bioavailability. Particularly higher Fe concentrations were found in the lung, liver and spleen than the remains. Lower Fe concentrat ions were recorded in skin and blubber. Hepatic Fe concentration positively correlated with maximum duration of dives among cetacean species ( y= 285. 17x 0. 5267 ; r 2 = 0. 92; y : average hepatic Fe concentrations in Lg#g - 1 dry weight; x : maximum diving duration in min) . Using this correlation, the maximum duration of dives of Dall. s porpoise was estimated as 11. 85 min.

Abstract:In this paper , the orthogonal experiment of three factors and three levels L9 ( 3 4 ) was adopted using blood corpuscle of Haliotis discus hannai which were in vitro immersing in PHA solution . There were three factors and three levels which included PHA treatment t ime : 6 h , 1 2h , 18 h ; PHA concentration : 0. 02%, 0. 04%, 0. 06% ; sampling time: 16: 00- 18: 00, 22: 00- 24: 00, 4: 00- 6: 00, respectively . The test s were repeated two times. The results showed that the optimal combination of three factors and three levels in the detection of triploid Haliotis discus hannai by using chromosome specimens from blood cells of living bodies was: PHA treatment time 18h, PHA concentration 0. 04% and sampling time 22: 00- 24: 00. The order of the three factors was PHA treatment time .. PHA concentration .. sampling time. Under the optimal combination of the three factors and three levels many clear, proper length and wel-l scattered metaphase chromosomes could be obtained. It is the best method of detecting the ploidy of living Haliotis discus hannai . Determining the relatively content of blood corpuscles.. DNA with flow cytometry could also make ploidy detection. But the device was expensive and the method was diff icult to be spread.

Abstract:Through several years.. research, we successfully obtained the progenies of the distant hybridization between Oreochromis aurea( .. ) and Sinipperca chuatai ( .. ) . Traditional morphological data and truss network data were combined to conduct multivariation analyses to study the morpho logical variations among O. aurea f rom American(Ao) , progenies of Ao( .. ) .. S. chuatai ( .. ) ( As1) , O. aurea from Egypt ( Eo ) , progenies of Eo( .. ) .. S . chuatai ( .. ) ( Es1) , as well as so..called back..crossing of the As1 ( As2) , and sister..brother crossing of Es1( Es2) . Ch..square analysis of merist ic characters indicated that there were no signif icant differences among these six experimental f ishes. But the results of cluster analysis of metric characters plus truss network data showed that there were some differences between these six experimental f ishes and especially, As1 and Es1 were quite different from the other fishes, though no difference was detected between As2 and Eo . Discriminant analysis indicated that there were significant differences among these six fishes. The identif icat ion accuracy was ranked as As1, Es1 and As2 [ 100%] > Es2 [ 96. 2% ] > O. aurea A [ 95% ] > O. aurea E [ 90%] . According to the results of mult ivariat ion analyses and subspecies 75% rule, the progenies of O. aurea .. S. chuatai are different from O. aurea. So it is reasonable to improve aquaculture strains using distant hybridization.

Abstract:Monsoon river prawn Macrobrachium malcolmsonii is an exotic freshwater prawn species recently introduced from Sri Lanka . The external appearance of the prawn is that of a spindle . The prawn consists of 20 pieces. Except telson , each piece has a pair of appendixes. Females and males have - shaped and Mshaped protuberances on the bases of f ifth walking legs. The surface of the gastric , cardiac and antennal areas of the carapace is rather coarse and there are several light brown mottles paralleled with prawn body on the both sides. During the fry ( 1- 3 cm) period, the body color of most individuals is light brown. Af ter sexually matured, the body color turns light brown, light blackish brown and light black. The tip of the rostrum slightly slants upward and extends forward, but neither going beyond the end of the antenna squama of the antenna seconda nor slanting higher than the tip of the coronal apophysis on the upper edge of the rostrum. The dental formula of rostrum is 7- 13/ 4- 6. Monsoon river prawn M. malcolmsonii belongs to the same genus as M. rosenbergii and M. nipponense. Though there are many similarit ies in their biological characteristics, there are also many differences in body color, rostrum, carapace and sternite pleurons, which can be obviously ident ified. This paper describes the external features and structures of M. malcolmsonii in detail, compares major characteristics of M. malcolmsonii and the other two species mentioned above. In addit ion, the questions are raised on the / male prawn specialization0 based on the second chelipod/ body length ratio.

Abstract:Based on f low..through method under the laboratory conditions, the energy budgets were measured and the bioenergetics models were described for seven species of fish distributed in the Bohai Sea and Yellow Sea ecosystem. As a result, the patterns of energy allocation for these seven species of marine f ish can be classed into three categories: ( 1) lower metabolic consumption and higher growth eff iciency, e. g . Schlegel.. s black rockfish and finespot goby; ( 2) higher metabolic consumpt ion and lower growth ef ficiency, e. g. black porgy and fat greenling; ( 3) both metabolic consumption and growth ef ficiency in medium level, e. g. red seabream, tiger puffer and chub mackerel. The significant difference of energy budgets in these seven species of marine fish is presumably related to the species distinction in their ecological habits. In comparison with f reshwater carnivorous f ish, the metabo lism energy in these seven species of marine f ish is higher than that of the average in the relative f reshwater species, indicating that the marine fish should be categorized as those of high metabolic consumption and low growth eff iciency.

Abstract:It is important to study the nutrition requirement of Takifugu obscurus because the puffer is a precious and potential species for intensive aquaculture .A 12-week growth experiment was conducted in indoor concrete ponds to determine the optimum protein level for the growth of juvenile T .obscurus , at the temperature of 24 - 26 ℃.White fish meal was used as the sole protein source to formulate isocaloric experimental diets at six protein levels (35 %, 40 %, 45 %, 50 %, 55 %, 60 %).Weight gain and feed efficiency of fish improved with increasing dietary protein level up to 50 % and reached a plateau above this level .Weight gain of fish fed 50 % protein diet was significantly higher (P <0 .05)than that of fish fed 35 %, 40 %and 45 %protein diet , but not significantly different from that of fish fed 55 % and 60 % protein diet (P >0 .05).The protein efficiency ratio tended to decrease with increasing dietary protein level , but there is no significant difference among the diets containing 35 %, 40 %and 50 % protein levels (P >0 .05).Dietary protein requirement using broken-line model was estimated at approximately 49 % and 46 % for maximum growth and maximum protein utilization , respectively .Therefore , it is suggested that the optimum dietary protein level for juvenile T .obscurus should be 46 %-49 %.The result is similar to that of tiger puffer T .rubripes .

Abstract:Feeding experiments were conducted with protein diets ( PD) and protein..free diets ( FPD) for juvenile Penaeus vannamei ( average body weight s, 0. 2627- 0. 2715g ) . The diets( PD) are prepared by casein and gelat in as protein source with high biological values. The requirements of juvenile Penaeus vannamei for essential amino acids ( EAA) were determined based on the daily depo sition and changes of each EAA in shrimp body. The minimum requirements of EAA needed to satisfy the requirements of juvenile Penaeus vannamei growth ( g- 1..( 100g body weight) - 1..d- 1) were Threonine 0. 046, Valine 0. 054, Methionine 0. 029, Isoleuline 0. 069; Leucine 0. 087, Phenylalanine 0. 051, Lysine 0. 086, Histidine 0. 025, Arginine 0. 097. When dietary protein levels was 40%, net protein ut ilization ratio was 50% and the feeding rate was 20% of the body weight, the minimum contents of EAA in the dietary ( g ..kg - 1 diet ) were as follow s, Threonine 11. 5, Valine 13. 5, Methionine 7. 3, Isoleuline 17. 3, Leucine 21. 8; Phenylalanine 12. 8, Lysine 21. 5, Histidine 6. 3 and Arginine 24. 3.

Abstract:The histopathology of the Meretrix meretrix Linnaeus suffering from red muscles disease was studied by histological and histochemical staining methods. The main characters of diseased clam are that the histological structures were irregularly changed, the epithelial cells swelled and detached. Some abnormal structures were observed in tissues of gills, mantles, digestive diverticulum, etc. , e. g. strong basophilic granules, acidophilic granules and protozoan. Histochemical studies, on the other hand, indicate that amount of amylose and activity of phosphatase appear obvious changes, e. g. the content of amylose increases in the absorptive cells of digestive diverticulum and intestine. The activity of alkaline phosphatase ( AKP) increases and the acid phosphatase ( ACP) decreases in the tissue of digestive diverticulum and digestive tract, while in the gill tissues, the activity of ACP increases and AKP decreases.

Abstract:Regulations of cultural conditions on the exotoxin expression of Vibrio mimicus HX- 4 from Eriocheir sinensis have been studied in vitro. Through measuring hemolytic and proteolytic activities of crude exotoxin in different cultural conditions. The results demonstrated that different media, NaCl percent, initial pH, soluble oxygen, cultural temperature and time all effected on the exotoxin production. The optimal conditions of the exotoxin expression are that the HX- 4 strain is inoculated in 0. 5% NaC-l BHI with init ial pH 8. 0 at 28 e for 36h in shaking cultivation or 48h in stat ionary cultivation. The pathogenic experiments of the crude exotoxin to mice and Eriocheir sinensis also have been finished. The results showed that the crude exotoxin is pathogenic to both of experimental animals, the higher the hemolytic titer of crude exotoxin is, the higher its lethal rate to experimental animals is. The crude solut ion of exotoxin with protease and no hemolysin has pathogenicity, but no lethality to experimental animals.

Abstract:An anti-viral protein was isolated from Megalobrama amblycephala serum by affinity chromatography on a column of Sepharose 4B-GCHV structure protein .Depending on exogenous β-mercaptanethanol , the isolated anti-viral protein could agglutinate rabbit erythrocytes treated by trypsin .Galactose was the most active inhibitor to the protein in hemagglutination inhibition test , and mannose had less activity than that of galactose .The molecular weight of the protein was 240 000 in PG-PAGE and SDS-PG-PAGE demonstrated that it had a subunit structure consisting of a single component with a molecular weight of 15 000 .The anti-viral protein was designated anti-viral galectin-like protein of M .amblycephala .After treatment with rabbit anti-serum, the rest serum of M .amblycephala lost it' s anti-viral and hemagglutination activity completely .The amino acid sequence of N-end of the galectin-like protein was Lys-Val-Asn-Leu-Asp-Glu-Lys-Cys-Pro-Phe , and no protein with the homologous amino acid sequence was found in the GenBank .

Abstract:The denaturation of myof ibrillar protein of bighead carp ( Aristichthys nobilis) during frozen storage at different temperatures ( - 10 e , - 20 e , - 30 e and - 40 e ) was studied by measuring the solubility of actomyosin, ATPase activities and - SH content of myofibrillar protein. The results indicated that the solubility of actomyosin, ATPase activities and-SH content of myofibrillar protein decreased as frozen storage progressed. The temperature of frozen storage can significantly inf luence the rate of denaturation of myofibrillar protein ( P< 0. 01) . The lower the frozen storage temperature, the slower the rate of denaturation of protein is.

Abstract:To observe the surface microstructure features by using scanning electron microscope , and study the surface antigen on lymphocytes of Paralichthys oliveaceus by form rosette with sheep red bloods cells and the cross-reactivity by using monoclonal antibody against human CD .Under scanning microscope , there are two kinds of lymphocytes , type I lymphocytes is characterized with being full of bumpiness and holes , the surface of type II lymphocytes is rather smooth with tiny finger like tubercle .The rosette formation of lymphocytes with sheep red blood cells is obvious however the rosette forming percents of lymphocytes is rather low, lower than 2 %.The cross reactivity of lymphocytes with monoclonal antibody against human CD showed that there are 5 .25 % 0 .38 % of NK cells in peripherial lymphocytes of Paralichthys oliveaceus , and the CD25 percentage of lymphocytes is 5 .25 % 0 .38 %.No cross-reactivity with T-cell antigen ,B-cell antigen and other activity antigen is found on the lymphocytes surface of Paralichthys oliveaceus .

Abstract:A pair of primers was designed based on the vp 28 gene and PCR was performed to amplify the gene f rom WSSV DNA. Insert ing the DNA fragment to the yeas-t E . coli shuf fle vector pPICZ and the recombinant plasmid ( pPICZVP28) that contains the target DNA fragment was obtained in the E. coli strain. The pPICZVP28 was introduced into Pichia pastoris strain X33 by electroporation. The transformant strain X33-1 was grown in BMGY media in fled f lask at 28 e . Induced by methanol for 72h, the samples of cell pellets and supernatant were collected by centrifugation and analyzed by SDS-PAGE and Western-blot, which confirmed that the strain X33-1 can express the WSSV. s envelope protein vp 28.

Abstract:Hard clams Mercenaria mercenaria were introduced from America to China in 2000. In this paper, the contents of 18 common acids, protein, fat as well as vitamines and mineral ions in edible part of hard calm were analyzed. The results showed that the contents of 18 common acids and the essential amino acid were 52. 42% and 40. 88% respectively and the average score of essential amino acid was 96. 4 .. 11. 3 according to the FAO/ WHO suggested level ( 1973) , and in which the smelled amino acid takes about 48. 30% . The content of protein in edible part of hard clam was higher( 60. 69% ) and the content of fat was much lower( 5. 60% ) than that of some bivalves. The results also indicated that the edible part of hard clam was also rich in amino acids, vitamins ( A, B1, B2, B5, B6) , and mineral ions such as Ca, Zn, Fe, Mn, etc. Hard clam was suggested to be with higher nutritional value which was good for human health.

Abstract:When the temperature runs up to 20 .. , the male Oreochromis nilotica w ill excavate holes on the sides or the bottom of the pool. Oreochromis nilotica will hide in the holes when scared by the fishing haul or other provocat ion. This would make the fishing operation very difficult , and the f ishing rate very low . With the ordinary seine net, the fishing rate of f irst haul is always under 30% , and the total fishing rate for the first three hauls is about 50%. On the basis of the study on the behavior of Oreochromis nilotica, a new fishing gear, a seine with a curtain net , is designed. On the newly designed seine, there is a curtain net between the sink line and the bolch line. The bolch line is much longer than the foot line, and will be loose enough in fishing operation to get into the hole to scare out the fish which is hidden in the hole. The newly designed seine was tested in four Oreochromis nilotica pools in Yantai and two Oreochromis nilotica poo ls in Taian respect ively. The results show that the f ishing rate is over 60% for the first haul, and the total fishing rate for the f irst three hauls is above 90% . The newly designed seine is more suitable for the fishing operation in the pool with harder bottom.