Abstract:The f ine structure of the gill in Pangasius sutchi has been investigated using transmission and scanning electron microscopy. The terminal of gill filament of Pangasius sutchi expands to be spoon-shaped. Many secondary gill lamellae arrange on two sides of each f ilament, and inlay each other. There are many taste buds distributing on the surface of gill arches and gill rakers. The respiratory surface of gill f ilament is rough and highly full of bloodvessels; the non-respiratory surface of gill f ilament is composed of tetragonal or hexagonal epithelial cells, which form various patterns. There are many holes among the circular ridgy patterns. The basic structure of the secondary lamellae consists of one or two outer epithelial layers, an intermediate basement membrane layer internal to which are the pillar cells and their f langes which line the blood channels. The chloride cells are frequently found in the base regions of the secondary lamellae and are overlain by a single epithelial layer , they often seemed to be open at the surface between adjacent epithelial cells. The relationships between the special structure of f ish gills and their funct ion are discussed.

Abstract:The ultrastructure of hemocytes of Haliotis discus was studied. Five types of hemocytes were recognized: large granule cell, small granule cell, special granule cell, transparent cell and lymphoid cell. According to the morphology of the nucleus and the characters of cell organelle in the cytoplasm, large granule cell and small granule cell are considered the same type cell in different developmental stages, however, special granule cell is early haemocytes of them. Transparent cell and lymphoid cell are completely differentiated cells.

Abstract:By using the polyacrylamide gel electrophoresis( PAGE) , this paper studies isozymes( LDH, EST, SOD) , haemoglobin and serum proteins of the inbreeding F1-generation gynogenetic grass carps、bisexual reproduction grass carps in the same ages and carps, respectively. The results of experiments show that there exist some differences in their electropherograms between the inbreeding F1-generation gynogenetic grass carps and bisexual reproduction grass carps; The biochemical characteristics of the inbreeding F1-generation gynogenet ic grass carps do not directly ref lect the influence by the genetic materials of the carps.

Abstract:Ages of grass carp ( Ctenopharyngodon idellus) larvae collected from Jiujiang section of the Yangtze River in 1998 , Shimatou and Shamao sections of the Yangtze River in 1999 , were determined by counting the increments in otolith. Birth date and daily growth rate of body length of the larvae were estimated based on the daily age. Average age of the larvae from Jiujiang , Shimatou and Shamao sections were 10. 7d , 8. 8 -16. 0d and 7. 0d , respectively. In the samples from Shimatou section , the age of larvae sampled on May 16 was older than that of the other samples. The average and specific growth rate of body length of the larvae from Shimatou section was 0. 11mm . d -1 and 1. 32 % , respectively. The correlation between body length and age was insignificant except in few samples , which suggested that the growth rate of larvae from the middle reaches of the Yangtze River was unequal , and some individuals might have suffered from the food absence.

Abstract:The growth of Mauremys mutica , Cuora trifasciata and Chlydre serpentina in one year is studied and compared. Under the condition of natural temperature and artificial feeding in Guangzhou , when Mauremys mutica was reared for 343 days , its average weight increased from 10. 3g to 71. 9g and its growth rate was 4. 37mg·g-1 ·d-1, Cuora trif asciata for 343 days , from 15. 9g to 67. 8g , its growth rate 3. 68mg ·g-1 ·d-1; Chlydre serpentina for 362 days , from 10. 8g to 263. 1g , its growth rate 5. 09 mg·g-1·d -1. Namely , the growth rate , Chlydre serpentina’s > Mauremys mutica’s > Cuora trif asciata’s was found. The relationship among mass , carapace length , carapace width and day old are very significant . The growth rate of turtles decreased in low temperature and the result was Mauremys mutica’s > Cuora trif asciata’s > Chlydre serpentina’s. Under the condition of same feeding, the individual growth rate was different. That was Chlydre serpentina ’s > Mauremys mutica’s > Cuora trif asciata ’s. The effects of feed translating were Chlydre serpentina ’s > Mauremys mutica’s > Cuora trifasciata ’s. The quantity of ration was Cuora trif asciata ’s > Mauremys mutica’s > Chlydre serpentina’s.

Abstract:Based on the surveyed data of water quality and phytoplankton in Hangzhou Bay , the eutrophication situation and the phytoplankton diversity etc. in the bay were analyzed by nutrient status index , diversity index and similitude index in this paper. The results show as follows. 1) The level of eutrophication decreased from the inshore to the offshore in the bay and the percentage of COD , DIN and DIP in the nutrient status index accounted for 64. 3 % , 34. 6 % and 1. 1 % respectively. 2) The distribution of diversity index of phytoplankton showed the characteristic of decreasing , increasing and then re2decreasing from the northwest to the southeast , and the axis of the lowest index was located near the A -A’line between 121°30′E, 30°00′N and 122°30′E, 31°00′N in the bay. 3) The diversity index of phytoplankton was closely related to the nutrient status index , and the offshore area was more likely to become the ”danger area”, which may result in red tide bloom because of their negative correlation in that area. 4) According to the growth environment for phytoplankton , Hangzhou Bay can be divided into inshore area and offshore area based on the A -A’line , and the inshore area was worse than the offshore area for phytoplankton growth. 5) According to the similitude index , phytoplankton community in Hangzhou Bay can be divided into inshore phytoplankton community and offshore phytoplankton community , and the boundary of their geographic distribution was the same as above.

Abstract:The mechanism of inhibitory gonadal precocity of GnRH-A and Zipi in cultured large yellow croaker was studied using pituitary histophysiological methods. The present result s indicate that it occurred vacuole in the cytoplasms of many gonadotropic cells in the pituitary of gonadal precocity, which shows that the cause of gonadal precocity may be due to entering the secretory activity of GtH cells in pituitary. The GtH cells of the cultured large yellow croaker, which had been fed with food containing GnRH-A, showed a weak immunopositive reaction to GnRH ant-i idiotypic antibodies, while those of the gonadal precocity in the control group showed a strong immunopositive reaction. These results indicate that responsive capability of GtH cells in the experimental group to GnRH-A are weakened, and desensitizat ion effect occurred. It may relate with GnRH-A inhibitory gonadal precocity of large yellow croaker. GnRH receptor on the membrane of GtH cell of the cultured fish, which had been fed with food containing Ziqi, did not showed desensitization phenomenon, which indicates the mechanism of inhibitory gonadal precocity of Ziqi differs from GnRH analogues.However, the definitive mechanism still needs further study.

Abstract:Monoclonal antibodys (Mab) raised against Vibrio alginolyticus strains (1. 1587) by that primed spleen cellsfusedwithSP2/0myelomacellsusingpolyethylingglycol. Therateoffusionwas 45.8%andtherateof positiveinfusionwas 77.4%byELISA. Mabs, AC1-C9,AC1-C11 ,BB4-D4,AD1-A3,AD1-F3,AD2E7 ,AD2 -F7 , were harvested using usable hybridomas clone twice by limiting dilution. Ascites with Mabs against Vibrio alginolyticus were harvested by injecting BALB/ C mice. The titer of the ascites was 1 : 1000. In comparison , Mab AD1 -F3 reacted with only four Vibrio alginolyticus strains . No cross2reactions were observed among thirteen non2Vibrio alginolyticus. Using this Mab ascites , an indirect Enzyme2Linked Immunosorbent Assay (ELISA) for rapid diagnosis of Vibrio alginolyticus has been developed. This method can detect 1 ×104 bacteria per ml with in 6 -7 h. After the method was optimized , 11 samples were detected for Vibrio alginolyticus.

Abstract:The endocrine cells in the digestive tract and pancreas of sof-t shelled turtle ( Trionyx sinensis ) were studied by using Streptavidin peroxidase ( S-P) , an immunohistochemical method with ant-i Glueagon ( Glu) , Gastrin( Gas ) , Somatostation( Som) , 5-hydroxytry-ptamine( 5-HT ) body . The result showed that 5-HT cells were distributed in the all digestive tract, in which the density of 5-HT cells in the small intestine were highest, that in the stomachus, large intestine, rectum secondly, and that in the oesophagus lowest. The density of Som cells in the stomachus fundus, pyloricus and small intestine were higher than that in stomachus cardia, large intestine, rectum and pancreas gland. The Gas cells distributed in the stomachus corpus, pyloricus and small intestine, there were not the Gas cells in the stomachus cardia, large intest ine and rectum. The Glu cells were distinguished in the pancreas gland and not in the intestine.

Abstract:By using hybridoma2monoclonal antibody technology , thirteen hybridoma cell lines which secrete monoclonal antibodies (Mab) against the IgM of European eel (Anguilla anguilla) had been established. And the characteristics of these Mabs had been described. In isotyping analysis , the results showed that among these Mabs , three of them are IgM , five of them are IgG1 , three of them are IgG2a and two of them belong to IgG2b ; all of these Mabs have the ability to bind the eel IgM by ELISA and their titer ranged from 104 to 107 , seven out of thirteen Mabs are able to recognize the heavy chain of the IgM on western blot. Further experiments proved that all of them only specifically bound to Ig of European eel and Japanese eel , and did not have any cross reaction with the Ig of Crucian carp 、Colossoma brachypomum cuvier 、Tilapia 、Clarias f uscus 、Ictalurus punctatus , and also did not react to the fish common bacterial pathogen , such as Aeromonas 、Edwardsiella 、Vibrio 、Flexibacter columnaris 、Salmonella and Escherichia coli . The measuring sensitivity of the Mab MaE9D7 to purified eel IgM was as low as 16ng. All of these results demonstrated that Mabs were highly specific and sensitive to the Ig of European eel , and had the great potential to be used for analyzing the structure of eel IgM , monitoring the immunity level of cultured eel , and pathogen diagnosis .

Abstract:Total RNA of leukocytes from orange-spotted grouper, Epinephelus coioides were extracted by Tripure kit. The first strand cDNA was synthesized by using MMLV reverse transcriptase. Then the cDNA were amplif ied by LD- PCR with SMART ó Oligonucleotide and CDS ó as Primers. After proteinase K digestion , Sfi . digestion, chromatography through CHROMASPIN- 400, over 500bp fragments were collected and ligated with KTripIEx2 vector. After package in vitro, cDNA library containing 1. 65 @ 106 private clones was constructed, in which more than 82% clones were recombinant . After amplif ication, the titer of the library was 7. 5 @ 109 pfu#mL- 1. The insert DNA were between 500 and 2300bp, and most of them were between 750 and 1000bp. The library can be used to screen cytokine genes of orange-spotted grouper.

Abstract:P31 gene of infect ious spleen and kidney necrosis virus ( ISKNV ) of Siniperca chuatsi had been sequenced. The open reading frame of the gene is 675 nucleotides long, encoding a putative protein of 225 amino acids with 25. 3kD. The 5. noncoding region has a TATA box and CAAT motif. A stem loop, a direct repeat sequence and a short dyad are contained downstream of the translation stop codon. The results of comparison of the homology with other iridoviruses p31 genes showed that ISKNV p31 gene has lower identities with others. Alignment of amino acid sequences and a phylogenetic tree of p31 genes of four iridoviruses have been constructed. The results indicated that ISKNV is different from the viruses of the genus Ranavirus and Lymphocystivirus.

Abstract:Ribonuclease III gene of infectious spleen and kidney necrosis virus( ISKNV) of Siniperca chuatsi had been sequenced. The open reading frame of the gene is 771 nucleot ides long, encoding a putative protein of 256 amino acids w ith 28. 9 kD. The 5. and 3. noncoding regions have TATA boxes. A palindrome is contained downstream of the translation stop codon. Compared w ith the RNase IIIs found in ISKNV and other species, the higher homology among ISKNV and the two iridovirus is present, and ISKNV has lower identity with other species, especially yeast and rhabditida.

Abstract:In this paper , a pair of primers were designed according to the nucleotide sequence of the ompII gene of Aeromonas hydrophila reported. With the specific primers , one target fragment about 1kb was amplified from the A . hydrophila genomic DNA via PCR. The target fragment was inserted into the linearized plasmid vector pRSET A. The nucleotide sequence of the inserted fragment , containing ompTS gene , was determined. The longest open reading frame (ORF) of 1068nt was identified and predicted to encode a 3552aa protein OmpTS with the molecular weight of 38. 9 kDa. Hydrophobicity analysis suggested that except for a potential 202amino2acid signal peptide , the protein was hydrophilic. The nucleotide sequence of ompTS gene showed 83. 5 % homology to the sequence of ompII gene. A Genbank search using the BLASTP program found that , the predicted amino acid sequence of the ompTS gene showed substantial identity with those of the outer membrane proteins reported in Escherichia coli , Klebsiella pneumoniae , Salmonella typhi , Serratia marcescens , Photobacterium profundum and Citrobacter freundii. Based on the amino acid sequence comparison , a high degree of amino acid sequence conservation at N2terminal was revealed. These analysis indicated that the ompTS gene is probably a novel structural gene that codes for OmpTS , which most likely forms pores in the outer membrane.

Abstract:The complete gene of major capsid protein ( MCP) of the recently isolated iridovirus from Rana tigrina was amplif icated using the primers corresponding to the two sides of the frog virus 3 ( FV3) MCP gene and was cloned into the pGEM-T easy vector. The MCP gene of the virus was 1 392 bp in size and had much higher identity to FV3 ( 98% ) , the type species of the genus Ranavirus , than to lympocystis disease virus ( 52% ) , the type species of the genus Lymphocystivirus . Comparison of the MCP sequence here w ith this of other vertebrate iridoviruses showed that the iridovirus examined here was a new member of the genus Ranavirus.

Abstract:The effects of freezing rate and storage temperature on Ca2ATPase activity and salt2solubility of myofibrillar protein in Silver Carp muscle were studied in this paper. The results show that the freezing rate might have an effect on freeze denaturation of muscle protein in round fish with intact muscle tissue to a certain degree , but almost no effect on protein denaturation of minced fish or surimi as muscle structure destroyed , no matter in which were cryoprotectants added or not. However, the frozen stored temperature showed the freeze denaturation of protein in round fish evidently and in minced fish and surimi as well , the lower the frozen temperature , the less the extent of freeze denaturation. It was also observed that the freez denaturation of myofibrillar protein in fish muscle could be efficiently reduced by the addition of cryoprotectants especially in of minced fish or surimi.

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