Abstract:technology. We determined that the titer of the primary cDNA library was 2.8 × 10⁶ pfu/mL, the recombinant efficiencies were over 99%, the insert size of the library was between 400 bp and 2 000 bp, and the tpfu/mL. One hundred single clones were selected randomly to screen for primary detection and a total of 95 high. The ESTs were into 70 unigenes, comprising 11 contigs and 59 singlets. When compared with the public database by Blast, 20 unigenes -value < 1e-5) to knownwere unknown genes (hypothetical protein), and 48 homology to any sequence. Several functional genes of Troponin I, Enolase, Anti-lipopolysaccharide factor and Lysozyme, were identified by homology comparison. The quality of this haemocyte cDNA library is of a high enough standard to obtain haemocyte-specific expression gene information. lts provide important information on functional genes and lay the groundwork for screening of molecular markers in .

Abstract:egg was studied using artificial fertilization, histology, ultrastructural technology methods etc. The observation showed that there is only one micropyle in shell of egg’s animal pole of belongs to the type of deep concave and short channel.At water temperature of 17sperm passed through micropylar apparatus at 3 s after insemination and entered ovum at 5 s. Sperm star light was formed at 10 s after insemination. After 15 min after insemination ovum was in late time of second meiotic division and second polar body appeared. Ovum was in telophase after 20 min and second meiotic division was formed. Male and female pronucleus was fused after 90 min. vum was in metaphase of first karyokinesis at 140 min after insemination. At 180 min after insemination first karyokinesis was over and second cell was formed. The results showed that in micropylar apparatus of mature egg there is no micropylar cell and other stuff, which is similar to that of most other teleost. The inner diameter of micorpylar apparatus is 2.42 µm and near to the diameter of sperm head. There are many spines in inner walls of anterior micropylar apparatus and holes are arranged in spiral way, which was one of the reasons for monospermy. This study aims to provide basic data for the research of artificial reproduction technology of .

Abstract:Wingless-type MMTV integration site familywas obtained from the transcriptome database) and its entire cDNA sequence was acquired by SMART-RACE was 1239 bp in length and encoded 355 amino acids. The WNT4 amino acids sequences were specific to the WNT family. Homologous analysis of the amino acids showed that Platynereis dumeriliiemi-quantative RT-PCR results showed no expressiontestes, gill and mantle tissues results indicated that resting stage and the lowest expression at the growing stage. The expression levels of were characteristically testis-predominant during the entire reproductive cycle. The multiple tissue expression characteristics imply that WNT4 plays various biological roles in . We can infer from the expression pattern in the gonads that WNT4 also contributes to the process of gonadal development and germ cell maturation.

Abstract:interferon that plays an important role in both innate and cell-mediated immune responses. Because of the low similarity with their mammalian homologues, IFN-γ in teleost fish has only recently been identified. In this study, the cDNA sequence encoding the IFN-γ molecule was cloned from the orange-spotted grouper, , and its gene organization was revealed. in response to Poly I:C stimulation was also examined. The IFN-γ cDNA sequence consists of 867 bp, which encodes a putative protein of 200 amino acids. The gene is 5-kb long and contains four exons, which are comparable in size with IFN-γ genes in other vertebrates. This IFN-γ was only . Sequence analysis revealed that the IFN-γ in the orange-spotted grouper contains the typical motif and a conserved nuclear localization site in the C-terminal end and displays an alpha-helix structure similar to mammalian IFN-γ. Real time PCR analysis results showed that the expression of IFN-γ increased in the thymus, head kidney, spleen, kidney, skin, and gills following the poly I:C stimulation, indicating its role against viruses in orange-spotted grouper.

Abstract:To study the molecular mechanism of the small peptide transporter PepT1-mediated protein digestion and absorption, a full-length cDNA sequence of the using RT-PCR and RACE techniques. The full-length cDNA sequence of had 2762 nucleotides, including 141 nucleotides at 5′UTR and 479 nucleotides at 3′UTR. Its open reading frame had 2142 nucleotides encoding a 713-amino-acid peptide. The were most similar to those of at 77.6% and 74.0%, respectively; the deduced amino acid similarities were 78.0% and 76.7%, respectively; but the gene sequence varied in similarity to other animals from to 59.1%, and the deduced amino acid similarities were 60.5%—61.6%. The encoded protein molecular weight was predicted at The PepT1 protein had 11 helix trans-membrane regions; its amino acid sequence was highly homologous to those of other vertebrates. Phylogenetic analysis showed that the as its closest neighbor mRNA assayed by real-time PCR were differentially expressed by tissue type; the highest expression was in the foregut tissue and the second was in the muscle. However, mRNA expression was relatively stable after incubation for 7 days. The C. idellus. Its expression was higher during the night and lower during the day. This work provides a theoretical basis for the molecular mechanism of protein digestion and absorption by the small peptide transporter PepT1, which mediates intestinal transport of small peptides

Abstract:The ciliated olfactory receptor neurons (cORNs) in the olfactory sac were dissociated from by enzymatic and mechanical methods. The whole-cell patch-clamp technique was used to determine the resting potential, action potential and voltage-gated ion channel current. The results showed that the cORN was a bipolar neuron, containing cell body, axon and dendrites with olfactory knobs and cilia. The resting and action potential peaks of ciliated olfactory receptor neurons were (–57 ± 6.37) mV and (38.30 ± 0.95) mV, respectively. In response to depolarizing voltage steps pulse stimulation from –60 ~ +80 mV, the transient inward current was activated between – pA at +40 mV. The present study provides a base theory for research in the electrophysiological mechanisms of sex pheromones with their receptors and the application of sex pheromones.

Abstract:The present study describes the morphology, histology and annual cyclical changes in the ovaries of the cultured silver pomfret, . The ovary development was divided into six stages, as were the changes in the oocytes of the female. Cultured populations have a similar ovarian structure, development process and breeding season to wild populations. However, the gonadosomatic index in cultured populations are obviously higher than that of wild populations, which infers that factors such as temperature, photoperiod and nutrition in cultured environments have a great effect on the development and maturation of the gonad in . The present study provides basic information for the artificial breeding of this species.

Abstract:, is completed in estuaries where they are exposed to frequent and abrupt changes in environmental salinity. Biochemical composition (carbohydrates, proteins and lipids) content and digestive enzyme (amylase, lipase, pepsin and trypsin) activity were determined in three developmental stages (gastrula, eyespot and pre-hatching stage) of eggs under hyper- or hypo-osmotic stress. Increases in pepsin and trypsin activity (<0.05), lipid content (<0.05) in the eggs of the control group were noticed during embryonic development. Total carbohydrate content and amylase activity increased at the eyespot stage (<0.05). The protein content during all three developmental stages decreased under hyper-osmotic stress while it increased under hypo-osmotic stress (<0.05). Total lipid content was unaffected by hyper-osmotic stress at any developmental stage, whereas it reduced significantly at the pre-hatching stage under hypo-osmotic stress (<0.05). Total carbohydrate content did not show any significant variation after osmotic stress. Hyper-osmotic stress reduced enzyme activity at the three developmental stages; however, hypo-osmotic stress enhanced it, with the exception of reducing amylase activity in the eyespot stage (<0.05) and trypsin activity in the pre-hatching stage under both hyper- and hypo-osmotic stress (>0.05).effect of salinity changes on digestive enzymes was only statistically significant during the gastrula stage (<0.05). We suggest that proteins and lipids are important structural materials of embryonic development and provide energy for this process. Osmotic stress changed the activity of digestive enzymes, which further affects the use of yolk substances, and consequently affects embryonic development. or low salinity in estuaries potentially affects the embryonic development of, especially under salt-water encroachment.

Abstract:was introduced to China from South America in 1991. The individuals chosen for this experiment were the F3 offspring of the largest scallop, in terms of shell height, bred from the years 2005 to 2007. Twelve adults were randomly selected from this population with a selection pressure of 5%, and 12 self-fertilizing, hermaphrodite families were established. The control group (L₀) was produced by mixed crossing of the F3 offspring of this scallop. Kung breeding and comprehensive evaluation values were evaluated in the self-fertilizing family and the control group by inspecting indices such as growth and survival of the offspring. The aim was to breed excellent self-fertilizing scallops and provide the material basis for genetic improvement of the stock. The results showed that there were no significant differences between the fertilization rate of the self-fertilizing families and the control group. However, the hatching rates were significantly higher( followed. In the larva stage, the daily shell length growth was significantly higher in B₃ and C₂ than in the control group, but was significantly lower in the other self-fertilizing families ( were significantly higher than the control group, yet they were significantly lower than the control up to 4 months old (and C₂ displayed significantly higher growth rates than the others in terms of shell height, shell width, and body weight (0.05). The comprehensive evaluation and breeding value of the four traits of the different family lines were B₃ were two excellent self-fertilizing families. In the 12 families, self-fertilization depression occurred to some extent in nine, one failed and two (B₃, C₂) exhibited superior growth and survival. We conclude that it is feasible to establish high quality self-fertilizing families with large-size shell height, improving breeding efficiency, and shortening the breeding period.

Abstract:to purify aquaculture wastewater in the lab. Different nitrogen forms removed from aquaculture wastewater and the dynamic distribution of four groups of bacteria involved in nitrogen cycling, ammonifiers (itrosobacteria nitrifiers (NB) and denitrifiers (DB), were evaluated. Our results revealed that the removal efficiency of nitrogen and chemical oxygen demand () performed by the combined treatment (I+M) was significantly higher than when used separately ( removal rate by the I+M treatment reached 50.83% and 62.38 %, respectively, significantly higher than I (39.55% and 51.17%) or M (40.78% and 53.31%) separately ( than that of treatment I (48.23%four groups of bacteria on the roots surface increased in certain extent compared with treatment M, and during the last period on the ceramic surfaces showed an overall downward trend, e.g. AB Principal response curve analysis (PRC) showed that turbidity,-N, TN contribute great to total difference, during the early and middle treatment was more efficient at removing nitrogen than treatment I or M, but the overall decreased greatly in the later stage with differences between I+M and I becoming small. This purifying method has some drawbacks and therefore requires further optimization to ensure its effects are long lasting.

Abstract:The Lidao reef zone is a typical artificial reef ecosystem for rare marine animal stock enhancement in the coastal sea of North China. The sea cucumber, Haliotis discus hannai, are the main enhancement species in the system. Based on the data of biological resources obtained from an investigation of the Lidao artificial reef zone in 2009, a balanced trophic model of the area was constructed using the Ecopath with Ecosim software package. The effects of uncertainty of input parameters and Ecopath analysis sensitivity were explored. Trophic flow and system attributes of the Lidao artificial reef system were analyzed. The ecological carrying capacity for were also predicted. The model consisted of 19 functional groups, which covered the main trophic flow in the Lidao artificial reef ecosystem. The results showed that the pedigree index of the model was 0.51, and the input parameters having the greatest effects on the output parameters were the production/biomass ratios () of small demersal fish groups. A –50% change in the production/biomass ratios in small demersal fish groups gave a sensitivity of the biomass (B) of the same group from 100% to 136%. The total system throughput was estimated to be 10 786.680 , 27% of which flowed to detritus and 17% of which is tranferred out of the ecosystem in the forms of fishing and sediment. Total net primary production was 4131.966 and the total energy transfer efficiency was 10.5%. The proportion of the total flow originating from detritus was 39%, and that from primary producers was 61, indicating that the energy flow was dominated by grazing food chain. The ecological carrying capacity was defined as the level of enhancement that could be introduced without significantly changing the major trophic fluxes or structure of the food web. The ecological carrying capacities for . This accounts for 31.72% and 26.15% of the biomass of each, respectively, meaning there is further potential for stock enhancement.

Abstract:of cage nets and protect juvenile crab was evaluated. At the same time, the impact of illumination and were photophobic: under bright conditions they preferred to in the four corners of the water tank. Under dark conditions, they were randomly distributed. Both bait and the black entice into the cage. The rectangular escape vents at the bottom of the cage net were 4 cm×3 cm, 4.5 cm×3 cm, 5 cm×3 cm and 5.5 cm×3 cm length by height, which corresponded to (the corresponding carapace length with 50% retention) of being 3.75 cm, 4.24 cm, 4.54 cm and 5.08 cm, respectively, and SR (the selection range) being 0.65 cm, 0.45 cm, 0.62 cm and 0.82 cm, respectively. has a linear relationship with the length of the escape vent, being about 92.3% of its length. Based on the minimum carapace length of sexually mature the minimum market-acceptable size standard and the experimental results, we conclude that the most size (

Abstract:basis of the data collected from October 2010 to January 2011 in the tuna longline survey, the soak time calculation models of every branch line in each operation were developed by both modes of hook retrieval. The soak time of longline gear divided into one hour interval for the quantity of hooks and the individuals of bigeye tuna (), respectively. The respective catch rates (CPUEs) of bigeye tuna and yellowfin tuna in each hour interval were calculated. The results showed that (1) both CPUE of bigeye tuna and yellowfin tuna presented increasing at first and then decreasing trend along the increase of soak time. The reason was the lure effect fluctuation of bait and the lose of hooked fish; (2) the quadratic curves can be fit the relationships between soak time and the CPUE of bigeye tuna, and yellowfin tuna; (3) the CPUE of bigeye tuna and yellowfin tuna was the highest when soak time was 9.9 h and 10.1 h, respectively. This study suggested that (1) the soak time of each hook lasted about 9.510.5 h in the tuna longline operation for improving the fishing efficiency and decreasing the bycatch; (2) the soak time of the longline gear could be considered as the effective fishing effort and used to standardize the CPUE.improve the fishing efficiency and to decrease the bycatch and will be for the references to the fishing strategy and CPUE standardization.

Abstract:s relating to farming are poorly understood. To assess the effects of co-cultured fish (grass carp, gibel carp, silver carp and bighead carp or silver carp and bighead carp) and formulated fish feed supplement (with or without feed supplement) on water chemistry characteristics, a 155-day field experiment was carried out in land-based enclosures. Four treatments including GGSB-F (co-cultured fish grass carp, gibel carp, silver carp and bighead carp were fed formulated feed), GGSB-NF (co-cultured fish grass carp, gibel carp, silver carp and bighead carp were not fed formulated feed), SB-F (co-cultured fish silver carp, and bighead carp were fed formulated feed) and SB-NF (co-cultured fish silver carp and bighead carp were not fed formulated feed) were established. The results indicated that the combination of co-cultured fish species could significantly affect Ca²⁺, total alkalinity (A) and total phosphorus (TP), while the feed supplement regime affected Ca²⁺, A, ammonia (TAN), TP and chemical oxygen demand (COD_Mn). Dissolved oxygen (DO) and Twere higher in the GGSB-F enclosures. Conversely, DO and SD were higher, but Ca²⁺were lower, in the SB-NF enclosures. During the experimental period, Ca²⁺ decreased, while TN, TP, TAN and COD_Mn increased. DO and SD were positively correlated with each other and negatively to the levels of TAN, TN, TP, Ca²⁺ and COD_Mn. Therefore, water qualityTAN and COD_Mn at the levels below the critical values, and maintaining Ca²⁺ and DO at high and constant levels.

Abstract:Based on monthly bottom trawl surveys in the central waters of Jiaozhou Bay from September 2008 to August 2009, a study on the community structure and diversity of shrimps was carried out using an index of relative importance, ecological diversity indices, multivariate analysis and other methods. Survey results indicated that 17 shrimp species, belonging to 15 families and six genera, were caught. The relative monthly shrimp resource, which was highest in September, October and June, varied dramatically among months. The average catch weight per haul (h) ranged from 223.9 g/h to 3 422.9 g/h, and the average catch number per haul varied from 41 ind/h to 1 608 ind/h. Monthly changes were observed in the dominant species Alpheus japonicus. There were severe changes in the diversity index among months, with the indices in spring and summer being higher than those in autumn and winter. The Margalef’s species richness index () and Pielou’s evenness index (0.76, respectively. MDS and CLUSTER analysis identified four month groups, (December, January, February and March), Group III (June, September and October) and Group (July, August and November), for shrimp communities during the entire year in this area. ANOSIM analysis revealed significant differences in species composition between and Group III or Group II and Group IV, but no significant between other groups. Typical species within-group and discriminating species between groups had a significant influence on the community structure. BIOENV analysis showed that bottom Jiaozhou Bay.

Abstract:A 42-day feeding trial was conducted to investigate the effects of dietary phospholipids on the choline requirement of the juvenile Chinese mitten crab, . Twelve purified diets were formulated containing two phospholipid levels (1% and 2%) and six choline chloride levels (0, 250, 500, 1 000, 2 000 and 4 000 mg/kg diet), and were fed to juveniles (0.22g±0.01g) with four replicates in each treatment. The results showed that dietary choline and phospholipids had a significant interaction on survival, weight gain, specific growth rate and feed conversion ratio (<0.05). Crabs fed diets with 1000 mg/kg dietary choline chloride (1% phospholipids) and 500 mg/kg choline chloride (2% phospholipids) obtained the highest survival rate, and significantly differed from that of other treatments with low choline chloride (<0.05). Crab weight gain and specific growth rate significantly increased with dietary choline levels from 500 to 4000 mg/kg (<0.05). Feed conversion ratio decreased significantly with dietary choline chloride increasing from 1 000 to 4 000 mg/kg (<0.05) at the 1% phospholipid level. At the 2% phospholipid level, feed conversion ratios of crabs fed diets with 500 and 1000 mg/kg choline chloride were significantly lower than those of the 0 mg/kg group (<0.05), but no significant differences were detected among the 250, 2 000 and 4 000 mg/kg groups. Whole body crude lipid hepatopancreas triglycerides, hepatopancreas total cholesterol, hepatopancreas glutamate pyruvate transaminase and glutamate aspartate transaminase activities significantly decreased with choline chloride increase, regardless of the phospholipid level (<0.05). Broken-line regression analysis based on weight gain against dietary choline levels indicated that the optimal dietary choline requirements of juvenile were 1 429.65 mg/kg and 529.65 mg/kg at the 1% and 2% phospholipid levels, respectively.

Abstract:The effects of increasing levels of dietary soybean lecithin (SL)on growth, ovarian development, tissue ultrastructure and fatty acid-binding proteins (FABP) mRNA expression(initial body weight 25.64 g±1.53 g), was investigated over 8 weeks. Five treatments were applied as 0% (Diet 1), 1% (Diet 2), 2% (Diet 3), 4% (Diet 4)2% SL had a significantly higher gonadosomatic index (GSI) than those fed with Diets 1 and 2 , and the hepatosomatic index (HSI) showed a decreasing trend with increasing levels of dietary SL>0.05). With increasing percentages of dietary SL, the hepatopancreatic lipid droplets increased in number and size and the structure and texture was more complete and evenly distributed. The ovarian lipid droplets and yolk granules also increased in number and size. Higher dietary SL yielded higher hepatopancreatic and ovarian FABP mRNA expression. Our results suggest that dietary SL has a positive effect on ovarian development in redclaw crayfishbroodstock, and at least 2% SL should be supplemented in broodstock diets to enhance ovarian maturation during aquaculture.