Abstract:As an important mariculture species in China, the rapid development of breeding industry of Japanese flounder (), lead to the breeding of Japanese flounder facing more and more serious problems at present, such as growth rate depression, fertility declined, resistance recession etc. In order to keep Japanese flounder aquaculture developing healthy and sustainability, it is necessary to get fast-growing Japanese flounder by crossbreedingand selecting. In this paper, four base stocks of Japanese flounder (), including Resistance stock (RS), Korea stock (KS), Resistance Japanese stock (RJ), Japanese Resistance stock (JR), were used to crossbreed and establish Japanese flounder families. The total length and body weight of these families would be measured, when they were about 19 months old. The SPSS system was used to analysis the data, at first. And then, estimated breeding values (EBVs) and other genetic parameters of total length and body weight were estimated by DMU package, based on the method of REML (Restricted Maximum Likelihood) and BLUP (Best Linear Unbiased Prediction) procedure. Results showed that, the phenotypic value of crossbreeding combination, KS (♂) × RJ (♀), was best among all the combinations. The heritabilities of Japanese flounder’s total length and body weight were 0.301 and 0.295, respectively, both of which were medium heritability. The correlations coefficient of breeding value and phenotypic value of growth traits were 0.838 and 0.827; both phenotypes of total length and body weight were statistically highly significant (<0.01) correlated to their breeding values, respectively. That is to say, the prediction result of breeding value reasonable accuracy was perfect. Comparing the means of breeding value of all combinations, we selected KS as male and RJ as female to breeding new species, because the two has the best breeding value relative to sex. By this method, new Japanese flounder variety growing rapidly could be brood successfully.

Abstract:It is important to consider the difference in contribution to reproduction between parents when assessing the effectiveness of enhancement programs. We used 18 microsatellite markers to estimate the of thirteen parents (five ♀, eight ♂), based on a sample of one hundred and eighty eight offspring of Japanese flounder ().When using 4 microsatellite markers, the PE was0.999 9 and accuracy was 92.02%. Using 6 microsatellite markers, the PE was 0.999 999 and accuracy was 96.81%. When using 8 microsatellite markers, the two values were 0.999 999 999 and 97.87%; using 10 microsatellite markers, the accuracy was 99.47%, and between 12–18 microsatellite markers, the accuracy was 100%. Our data suggest that the accuracy increased with an increase in the number of microsatellite markers. These SSR markers can be used for parentage determination and evaluating the effectiveness of enhancement releases. The results of PE testing suggest that paternity test accuracy of 100% can be achieved. Although all thirteen parents contributed to reproduction, there was considerable variation in contribution among individuals. The highest and the lowest parental contributions were 47.34% and 0.53%, respectively. The reasons driving these differences require further research.

Abstract:Güther to clone the mature peptide domain of the gene. The mature peptide consisted of 70 amino acids, including four functional domains B-C-A-D. The mature peptide fragment was subcloned into the prokaryotic expression vector pET-28a to construct a recombinant plasmid that was transformed into BL21 (DE3). Induction of the plasmid by IPTG yielded a special fusion polypeptide containing 6 at the N-terminus. SDS-PAGE analysis revealed that the IGF-I polypeptide had a molecular weight of 11.4 kD. The recombinant IGF-I protein proximately accounted for 58.5% of the whole bacterial protein 3 h post induction with IPTG. Thus,. Western-blotting analysis suggested the fusion polypeptide exhibited antigenicity to 6 antibodies. The IPTG-induced bacterial precipitate was denaturalized using 6 mol/L guanidine HCl, purified using Ni-NTA affinity chromatography, and annealed by gradient dialysis in urea, thereby yielding the purified protein. The recombinant IGF-I protein promoted the proliferation of breast cancer MDA231 cells, suggesting it is biologically active. Our results provide a basis and suggested methods for regulating the growth of farmedGüther.

Abstract:Prolactin is a single peptide hormone secreted from the animal pituitary. It has a varietis thought to be one of the primary hormones regulating the balance of the osmotic pressure and ions. PRL exerts its biological effects by binding with the prolactin receptor (PRLR). Thus, our understanding of the physiological functions and mechanism of action of PRL relies on a thorough understanding of the PRLR. Prolactin receptor is a single . mRNA has been detected in several organs associated with osmotic regulation. Changes in the concentration of ions or salinity in the environment are associated with changes in the expression of , which is thought to represent the molecular mechanism of PRL regulation.and the tissue specific expression pattern in . The full-length cDNA encoding using homology cloning and RACE PCR. The terminal UTR, 1 821 bp encoding region, and 406 bp 3’terminal UTR. Alignment of the deduced amino acid sequences of the and those of other vertebrates demonstrated that they shared many protein features common in fish. Phylogenetic analysis revealed that the putative PRLR amino acid. For example, the PRLR amino acid sequence of shared 87.25% similarity. For other species, the similarity was 86.86%57.43%49.16%T. rubripesReal-time quantitative PCR suggested that the he PRLR transcript was detected at a high level in gill, kidney, and intestine suggesting that in is in osmotic regulation. in the gonads, liver, and spleen, suggesting that prolactin has a range of physiological functions in fish. Our results have provided valuable insight into the mechanisms of var.

Abstract:is an economically important, cold, freshwater species in China. The species is cultured because of its taste and high protein value. Knowledge of the ontogeny of the alimentary canal and accessory glands is essential for understanding the nutritional physiology of larvae and juveniles. larvae reared under culture conditions based on histological observation. The larvae were held in one of three 0.3 m³circular tanks (12–16.2)for 185 d and fed an artificial diet. Based on the structural changes in the digestive system, we identified three distinct stages during embryonic development: (1) 0–552embryo initiated the intestinal phase of development, the original base cells of the digestive tract were localized below the chordoma, in a single layer of flat cells. During the second stage, the original base cells ofinner layer cells of the digestive tube proliferate, and the outer layer forms as two layers of cells. At the end of the embryonic period, the hatched larvae have an oil-rich yolk sac, a simple digestive system that resembles a blind tube, and lack of external connection in the anus. Between 2–7 days after hatch (DAH), the digestive system begins to rapidly differentiate but larvae remain entirely dependent on endogenous nutrition from the yolk sac. At the digestive tract was fully differentiated into the buccopharynx, esophagus, non-glandular and glandular stomach, and anterior and posterior intestine. By 14 DAH, larvae were transitioning from being dependent on endogenous nutrition to being exotrophic. The yolk sac was completely absorbed. At 49 DAH, the pyloric caeca was completely differentiated, and the liver and pancreas were functional. Between 64–105 DAH, the oropharyngeal cavity and stomach developed to the adult form with 36 pyloric caeca finger branches. The optimization of formulated feeding during larval culture of 14 DAH. Feeding during this period could be

Abstract:based on scale analysis. We collected 397 specimens from Long-xian, Shanxi, between 2009 and 2012. The scale annuli were characterized as the 3 year-old individuals accounting for 85.64% of the total samples. There was no significant difference in body length () between females and males and the Von Bertalanffy growth model was =729.38[1) was 0.08. The body weight growth inflexion point was at 13.10 years old, with a corresponding body length of 483.66 mm and body weight of 1 857.86 g. The results show that is a species of slow growing fish and it characterizes isometric growth and individual miniaturization in the natural reserve. We should take artificial propagation and artificial releasing to enlarge and revive its populations. The study is intended to realize its growth rhythm and take reasonable actions to protect and vitalize

Abstract:Auditory brainstem response (ABR) technic has been widely used to study hearing ability in fishes. In contrast to the behavior and traditional electrophysiology methods, this approach provides the following advantages: (1) there is no harm to the animals because it is non-invasive; (2) hearing sensitivity in untrained fish can be determined rapidly; (3) individuals are reusable. Using ABR approach, auditory evoked potentials of 10 Chinese suckers were recorded using two subcutaneous electrodes. We found that the fish can detected tone bursts from 100 to 5 000 Hz. The results showed the best hearing between 100 to 2 000 Hz, and the lowest threshold was 69.8 dB at 800 Hz. There was a positive correlation between the amplitude of the ABR waveform and the stimulus intensity, while there was a negative correlation between latency and stimulus intensity. Threshold values from 10 individuals were averaged to produce audiogram. Similarly to the most audiograms, it is a U-shape with thresholds increasing at both high and low frequency limits of hearing. The results have an important significance in protecting Chinese sucker and providing supporting data for access the effect of noise on fishes.

Abstract:We evaluated the effect of ferritin on the ability of the swimming crab to adapt to salinity stress. We amplified the gene from the sixth gill of the swimming crab and cloned it into the pET 28 a (+) expression vector. The vector was DE3 (BL21) and induced by IPTG. The erritin protein was successfully induced by IPTG, yielding a band that was consistent with the theoretical value (19.448 kD). was higher than that of empty vector transferred cells. Furthermore, the recombinant plasmid significantly improved the tolerance of Our results suggest that P. trituberculatus.

Abstract:during selection over four-generations for shell height using nine microsatellite loci. The mean effective number of alleles per locus () ranged from 1.435 to 2.096 among the four JCS generations. The mean allelic richness (odecreased from 0.492 to 0.269, suggesting there was a decrease in genetic diversity. In general, the allelic frequency remained stable over the four generations, but fluctuated up and down, apparently randomly, between generations. For example, the frequency of the HHM20-158 bp allele changed regularly from mid to high levels. Our results suggest that persistent selective breeding resulted in reduced genetic diversity and somewhat stable genetic structure in pearl oyster.

Abstract:We evaluated the effect of selection for one or two traits on the adductor weight of the bay scallop, population that had been produced by continuous progeny selection. We tested the effect of selection for shell height (selection pressure =10%) and/or body weight (selection pressure =1%). There was no significant difference(>0.05) in spawning, fertilization rates, hatching rates, the growth rate of larvae, the survival to the mid-cultivation period, and the morphology of adults between the positive selection group, negative selection group, and control group. However, there was a significant effect (<0.05) on the survival rate of juveniles. The survival during the grow-out period, the body weight of adults, and the values for each of the traits in the positive selection group were highest in the positive selection group and lowest in the negative selection group. We observed high realized heritability in both the positive and negative selection group, though the value was higher for the positive selection group than the negative. The inheritance of body weight and adductor weight was higher when both traits were selected than a single trait. Our results suggest there is a genetic correlation among shell height, body weight, and adductor weight, such that selection for the two former traits will affect the adductor weight. Concurrent selection for the two traits appears to have an additive effect, which will likely increase the rate of selection for adductor weight and thereby increase the yield. Our results provide a theoretical basis for improved selective breeding of the scallop.

Abstract:We evaluated the effect of light intensity on the specific growth rate (SGR), food intake (FI), food conversion efficiency (FCE), and the energy budget of red and green variants of the sea cucumber . We reared 96 green (6.28 g ± 0.02 g) and 96 red (6.34 g ± 0.04 g) individuals under six light intensities (0, 50, 300, 1 000, 2 000, or 3 500 lx) for 60 d at 16. Light intensity had a significant effect on specific growth rate (SGR) (3 500 lx, there was a negative relationship between the SGR of sea cucumber and illumination intensity. Both red and green variants grew fastest (<0.05) were observed in individuals reared under 50 lx. Stronger light intensity inhibits FI and reduces the efficiency of food conversion. The lower FCE under stronger intensity lighting may be function of the oxygen consumption rate (OCR). Our results suggest that OCR is significantly affected by light intensity (<0.05). The minimum OCR was observed in animals reared under 50 lx, suggesting that little energy was expended on respiration, with most being invested in growth. Energy metabolism was significantly affected by light intensity ( allocate 7% for growth and 90% for respiration and production of feces. As light intensity increases, the energy allocated to growth decreases and is instead diverted to respiration and production of feces.

Abstract:We developed a method for evaluating the drug residue risk in fishery products. Using a power model, we applied common hypothesis-testing approaches and confidence interval criteria to assess the dose proportionality of ciprofloxacin (20 in the intestine, liver, plasma, and muscle tissue of Linn. The residue in the liver tissue was proportional to the initial dose but there was no relationship between initial dose and the residue in the intestine, plasma, and muscle. The fitting formula for AUC and dosage was =0.901). Our results suggest that the . Our method represents a novel approach to assessing the drug residue risk in fishery products.

Abstract:Polymeric immunoglobulin receptor (pIgR) is a key immune factor, medicating the transport of polymeric immunoglobulins (pIg) through the epithelial cells into the mucosal secretions to protect the organisms from was first cloned and sequenced by pIgR contained 1 584 nucleotides, 3’UTR, 5’UTR, and an open reading frame (ORF) of 1 005 nucleotides that encoded for a polypeptide of 334 amino acid. Gene homology comparisons showed that Takifugu rubripes, indicating a high conservation of gene. Multiple sequence alignment demonstrated the pIgR of teleosts was composed of two Ig-like domains (ILDs), corresponding to the ILD1 and ILD5 of Mammal’s pIgR. The phylogenetic tree sequences of . Expression analysis revealed that pIgR gene was expressed in almost all tissues of healthy s, with higher levels in the mucosa-associated lymphoid tissues. were evaluated following bath immunization by real-time PCR, and the results showed that the relative expression amount increased firstly and then decreased within 72 h in all the tested tissues, and reached its maximum expression within 48 h and the appeared earliermucosa-associated. These results suggested pIgR played a critical role in the mucosal immunity of teleost.

Abstract:affinity chromatography, SDS-PAGE, western-blotting, antifungal assays, microscope and electron microscopic observation. activity at concentrations of 0.1–0.8 mg/mL. Furthermore, microscopic observation suggests that A. niger s. Electron microscopic analysis of indicated that the sporangium were atrophied, and the conidiospores were gathered into gouts. In addition, the hyphaes were cracked, contorted, and shortened, and the conidiosporangia were not visible. Taken together, our results suggest that hemocyanin possesses novel immune function (i.e., properties) likely related to inhibition of spore and hyphae growth.

Abstract:Recent outbreaks of “sleeping disease” have caused mass mortality in cultured mud crab and widespread economic loss in southeast China. Two novel pathogens (Mud Crab Reovirus, MCRV and Mud Crab Dicistrovirus, MCDV-1) have been isolated from crab exhibiting “sleeping symptoms”. Thus, there is an urgent need for the accurate and early detection of this pathogen to facilitate disease control. We developed a duplex nested-PCR protocol for the detection of MCRV and MCDV-1 in . Two pairs of primers targeting the conserved regions of the two viruses genes were designed using were ReoF (5′-GCAAATTGAACTACTACTACTTA-3′) and ReoR (5′-GATTCCTATTGTCAACTATCTCA-3′), and the inner primers were ReoNF (5′-ACTCATAGAGCAGTCATGGG-3′) and ReoNR (5′-ATATCGTCAGAATGTC GTTC-3′). The outer primers for MCDV-1 were DicF (5′-GCACTGGGTACTCTTCCTG-3′) and DicR (5′-AC ACCTACCAAAGCCCTAC-3′), and the inner primers were DicNF (5′5′-GGATACTATGGATGATGTTTC-3′) and DicNR (5′- ACAAAATACCAGATAAAGCAA-3′). The PCR products were differentiated by size. The first PCR was carried out using a PCR mix consisting of 1 μL DNA, 1 μL MCRV outer primer (ReoF/ReoR, 0.5 µmol/L), 0.4 μLO (to a final volume of 20 μL). The thermal cycle was: 94 for 10 min. The nested PCR was carried out using the same mix with 0.2 μL template DNA (the product of the first PCR), 0.8 μL MCRV inner primer (ReoNF/ReoNR, 0.4 µmol/L), and 0.8 μL MCDV-1 inner primer (DicNF/DicNR, 0.4 µmol/L). The conditions were the same as in the first PCR, except theannealing temperature was 53. We evaluated the specificity and sensitivity of the duplex nested-PCR. The assay did not exhibit any cross-reactivity with abalone shriveling syndrome-associated virus (AbSV), acute virus necrobiotic virus (AVNV), infectious hypodermal and hematopoietic necrosis virus (IHHNV), fish nervous necrosis virus (NNV), turbot viral reddish body iridovirus (TRBIV), or white spot syndrome virus (WSSV). We used a standard recombinant plasmid for each virus in the and MCRV/MCDV-1 served as the position control. The detection limit was 10¹ copies of the viral genome for the two viruses. Our results suggest that this technique is a rapid, reliable, and cost effective tool for identification of crab virus with high sensitivity, high specificity. The assay could also be applied as a rapid diagnostic tool in clinical samples in which MCRV or MCDV-1 infection is suspected and differential diagnosis is required.