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1.
Major histocompatibility complex class II antigens are important in vertebrate immune system.In the present study,the full cDNA sequence of class II A gene was synthesized by RACE-PCR from half-smooth tongue sole(Cynoglossus semilaevis),and its open reading frame(ORF) polymorphism was studied.The whole cDNA sequence was 992 bp in length,including the ORF with 717 bp.Twenty-five alleles were identified and clustered into two distinct groups according to the specific nucleotides/amino acids in specific positions.Eleven alleles belonged to Cyse-DAA while fourteen alleles belonged to Cyse-DBA.Four Cyse-DAA alleles were observed in one individual,and three to five Cyse-DBA alleles were observed in each of the three detected individuals,which indicated that at least two loci existed in each gene.Moreover,in order to study the function of the alleles in resistance to infection,200 individuals were intraperitoneally injected with Vibrio anguillarum and the first 20 dead individuals and 20 surviving ones were selected for genotype analysis.Fifty-six alleles were identified among the 40 individuals.Twenty-nine alleles belonged to Cyse-DAA and the other 27 alleles belonged to Cyse-DBA.Eighteen alleles were selected for studying their function in resistance to infection.Alleles Cyse-DAA*0201,Cyse-DAA*1101,Cyse-DBA*0401,Cyse-DBA*1102,Cyse-DBA*1801 and Cyse-DBA*2201 were identi-fied only in surviving individuals,while alleles Cyse-DAA*0901,Cyse-DBA*1101 and Cyse-DBA*1401 occurred more frequently in dead individuals.This study confirmed the existence and polymorphism of two class II A genes as well as the relationship between alleles of class II A genes and disease susceptibility/resistance in half-smooth tongue sole.  相似文献   

2.
A cultured female half-smooth tongue sole (Cynoglossus semilaevis) was crossed with a wild male, yielding the first filial generation of pseudo-testcrossing from which 200 fish were randomly selected to locate the Vibrio anguillarum resistance trait in half-smooth tongue sole at its microsatellite linkage map. In total, 129 microsatellites were arrayed into 18 linkage groups, ≥4 each. The map reconstructed was 852.85 cM in length with an average spacing of 7.68 cM, covering 72.07% of that expected (1 183.35 cM). The V. anguillarum resistance trait was a composite rather than a unit trait, which was tentatively partitioned into Survival time in Hours After V. anguillarum Infection (SHAVI) and Immunity of V. Anguillarum Infection (IVAI). Above a logarithm of the odds (LOD) threshold of 2.5, 18 loci relative to SHAVI and 3 relative to IVAI were identified. The 3 loci relative to IVAI explained 18.78%, 5.87% and 6.50% of the total phenotypic variation in immunity. The microsatellites bounding the 3 quantitative trait loci (QTLs) of IVAI may in future aid to the selection of V. anguillarum-immune half-smooth tongue sole varieties, and facilitate cloning the gene(s) controlling such immunity.  相似文献   

3.
Studies were conducted to determine the cause of the acute mortality of half-smooth tongue sole Cynoglossus semilaevis Günther juveniles in a fish farm in Jimo, Shandong Province, China, in June 2006. Gross signs of the diseased tongue sole included several petechiae and ecchymoses on the body and fin necrosis and hemorrhagic lesion at the base of the fin. Bacteria were isolated from kidney, liver and hemorrhagic lesions of the diseased tongue sole. Among14 strains, SJ060621 was proved to be highly virulent to juvenile tongue sole with LD50 value of <1.0×105 colony forming units (CFU) mL−1, while the remaining 13 were avirulent. Among the 16 antibiotics tested, SJ060621 was sensitive to gentamicin and nitrofurantoin. It was identified as Listonella anguillarum with conventional plate and tube tests in combination with API 20E analysis. 16S rRNA gene and partial HSP60 gene sequenceing analysis revealed that the strain was highly homologous with L. anguillarum. Examination of the infected musculature by electron microscopy indicated numerous bacteria and lots of macrophages containing phagocytosed bacteria. Histopathological investigations revealed severe necrotic degenerative changes in the infected organs. Indirect immunofluorescence assay (IFA) was employed to detect the location of occurrence of bacteria, and bacteria were found in aggregations in the inflammatory areas in musculature.  相似文献   

4.
Tissue expression and stock variation of isozymes of stone flounder (Kareius bicoloratus) were analyzed with horizontal starch gel electrophoresis. For the fourteen enzymes assayed, 31 loci were recorded. The results indicated that all the isozymes ex- amined were obviously tissue-specific. The expressions of SOD*, GDH*, G3PDH-2* and ADH-2* were detected only in liver, SDH-1*, MDH-1* and ADH-1* only in muscle, and LDH-B* and LDH-C* only in eyes. In comparison, MDH-2*, GPI-3* and SDH-2* were detected in all tissues examined. Other loci examined were detected in a variety of tissues. Muscle and liver were se- lected to detect the isozyme variation of the two geographic stocks of Qingdao and Weihai, Shandong Province, China. The percent- ages of polymorphic loci (P0.99) were 29.17% and 25.00%, the observed heterozygosities (H0) were 0.028 ± 0.014 and 0.040 ± 0.019, and the expected heterozygosities (He) were 0.039 ± 0.017 and 0.052 ± 0.022 in Qingdao and Weihai stock, respectively. The coeffi- cient of gene differentiation (Fst) and genetic distance (D) between the two stocks was 0.012 and 0.0011, respectively, indicating that the genetic differentiation is low between them. Compared with other species of Pleuronectiformes, both the percentage of polymor- phic loci and the mean heterozygosity of K. bicoloratus were at a middle level.  相似文献   

5.
The characteristics of a bacterium strain M3, isolated from cultured flounderParalichthys olivaceus with remarkable external sign of skin ulcer during an epizootic outbreak, indicated that the bacterium belonged to the speciesVibrio anguillarum. Challenge by I.M. (intramuscular injection), bath, and oral administration with M3 showed that it was highly pathogenic forParalichthys olivacues. The LD50 dose was 5.144×103 CFU/ per fish infection by I.M. injection. Recovered inoculated bacteria from the surviving fish revealed that the asymptomatic carriers could be a latent contagious source. Study of the effect of bacterial culture CFS (cell-free-supernatant) showed that the exotoxins produced by M3 play an important role in its pathogenicity for flounder. The resistance of M3 to 36 out of 41 antibiotics indicated that the bacterial disease outbreak was mainly attributable to the frequent and excessive use of antimicrobial agents; and that vaccination would be an effective precaution against bacterial disease. Contribution No. 4107 from Institute of Oceanology, Chinese Academy of Science. Contribution No. 270 from Experimental marine Biology Laboratory, CAS. Funded by projects under the Major State Basic Research Development Program, Grant G1999012003.  相似文献   

6.
Endocrine disrupting chemicals (EDCs) are increasingly viewed as persistent pollutants, similar to natural hormones in function. This paper describes the expression profiles of 7 genes (DMRT, VTG, GnRHR, FSHR, CYP17A, CYP19A, and CYP19B) involved in sex steroid synthesis and action as well as sexual development in adult male and female Cynoglossus semilaevis, after exposure to different concentrations of Bisphenol A (BPA) and 17β-estradiol (E2). Both BPA (1, 10, 50, 125, and 250 mg/kg) and E2 (0.5, 5, and 10 mg/kg) induced changes in target gene expression, although the estrogenic effects of E2 as a model estrogen were stronger. Among the 7 genes, VTG, CYP17A and CYP19 responded strongly to BPA or E2 exposure and can thus serve as reference biomarkers for estrogenic EDCs exposure in marine teleosts. These data will provide a window to establish a hypothalamic-pituitary-gonadal model in C. semilaevis to better understand the effect pathways of EDCs.  相似文献   

7.
Vibrio anguillarum is an important bacterial pathogen of aquatic organisms and a significant problem in aquatic farming. The rapid detection and identification of V. anguillarum, and other pathogens that infect marine organisms, is crucial to effective disease management. In this study, we developed a loop-mediated amplification (LAMP) assay to detect V. anguillarum in an hour in a single tube without the need for thermal cycling. Conserved regions of the metalloproteinase (empA) gene of V. anguillarum served as the targets for primer design. A fragment of the empA gene was amplified at 65°C in the presence of the primer mixture and Bst DNA polymerase. In the optimized LAMP assay, 6.7 pg of V. anguillarum DNA could be detected. Six strains of V. anguillarum and 17 strains of non-V. anguillarum bacteria were used in this study to evaluate the species specificity of the primers. The six V. anguillarum strains gave a positive result in the LAMP assay. This method was also validated in V. anguillarum-infected fish. This LAMP method is more sensitive than PCR in the detection of V. anguillarum and shows good species specificity. The LAMP assay is therefore an effective method for the quick detection of V. anguillarum both in the laboratory and in the field.  相似文献   

8.
Allelic variation in a total of 7 microsatellites was examined between elvers of freshwater eels （Anguillajaponica and Anguilla anguilla）. The number of alleles at these loci ranged from 8 to 26. A single test of each locus revealed significant deficits of heterozygotes （P〈0.01）. Significant departure from expectations of Hardy-Weinberg equilibrium （HWE） was found for all loci within four subpopulations of A. japonica, which opposes the panmixia hypothesis of Schmidt. Also exact tests of population differentiation based on allelic frequency distribution disagree the hypothesis of random distribution of individuals among populations. Population structure among four populations of A. japonica was revealed with FST value of 0.009 8 （P=-0.000 48; 10 000 iteration）. Pairwise matrixes of FST and RST showed a significant difference between two distantly related species-A, japonica and A. anguilla. Divergent time of the two species calculated by Goldstein method is over 2 million years. The results may challenge the Schmidt＇s theory about the distribution of freshwater eels.  相似文献   

9.
The International Eq一lation of State of Seawater,1980 and the PraeticalSalinity Scale,1978 have been adoPted by theUNESCO江CES沼COR八APSOJoint Panel ono‘eanogral,hie Tables and Standards(JPOTS),and endorsed bythese organizati6ns(Miller‘〕and Poisson,1981;Uneseo,1981).Th已new equa-tion and the Praetieal Salinity Seale are to be used for all values Published fromJan .1,1982 .The new equation 15 aeeurate for use in all oeeanie surfaee waters,but eannot be aeeurately aPPlied to…  相似文献   

10.
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11.
We cloned and sequenced a prtV-like gene from Vibrio anguillarum M3 strain. This prtV gene encodes a putative protein of 918 amino acids, and is highly homologous to the V. cholerae prtV gene. We found that a prtV insertion mutant strain displayed lower gelatinase activity on gelatin agar, lower protease activity against azocasein, and lower activity for four glycosidases. This prtV mutant strain also had increased activity for two esterases in its extracellular products, as analyzed by the API ZYM system. In addition, the prtV mutant strain exhibited decreased growth in turbot intestinal mucus and reduced hemolytic activity on turbot erythrocytes. Infection experiments showed that the LD50 of the prtV mutant strain increased by at least 1 log compared to the wild-type in turbot fish. We propose that prtV plays an important role in the pathogenesis of V. anguillarum.  相似文献   

12.
The Chinese tongue sole (Cynoglossus semilaevis) is widely cultured in the coastal region of East Asia and has excellent economic value. However, the high albino rate of the breeding population has caused a significant loss to the aquaculture industry. To study the molecular mechanism of albinism, the present study used an albino Chinese tongue sole family to construct three simple sequence repeat (SSR) linkage groups, and draft a preliminary linkage map related to albinism. After albinism-related loci mapping, 18 albinism-related loci were detected under two models (containing 2407 genes) compared to the Chinese tongue sole genome. One of these loci, the tyrosinase related protein (tyrp2), which has been reported previously as an important gene regulating both eumelanin and phaeomelanin levels, was indicated to be the possible cause of albinism. Thirty-five Gene Ontology (GO) terms and 14 Kyoto Encyclopedia of Genes and Genome pathways were annotated via bioinformatic analyses. One GO term with protein tyrosine kinase activity, which contained 10 genes, was previously suggested to affect fish albinism. These results establish a foundation for further in-depth study of albinism in Chinese tongue sole.  相似文献   

13.
Polyc·lonal antibodies raised against LHC II isolated from SDS-solubilizedBryopsis corticulans thylakiod membranes by SDS-PAGE, were characterised by double immunodiffusion, Rocket immunoelectrophoresis and antigen-antibody crossed immunoelectro-phoresis assays showed the antibodies had strong cross-reaction with allB. corticulans LHC II components (even with those which were incubated in boiling water) and showed immunological cross-reactivity with LHC II polypeptides of spinach and the marine green algaCodium fragile. The results suggested that LHC II of different species had similar antigenic determinants and also conservation of amino acid sequences of the polypeptides during evolution, and that the antibodies could cross react with apoproteins of D2 proteins (which contain P680) fromB. corticulans, spinach andC. fragile, but not with apoproteins of P700 Chl-proteins. Our results indicated some similarities in primary structure between LHC II of different species, and between LHC II and D2 proteins of marine green algae and spinach. Our finding that D2 and P700 Chl-proteins are not immunologically related suggested that P700 Chl-proteins and D2 proteins pass through independent evolutionary pathways. Contribution No 1717 from the Institute of Oceanology, Academia Sinica, This subject was supported by the National Science Foundation of China.  相似文献   

14.
Heavy metal pollution can affect the immune capability of organisms.We evaluated the effect of cadmium(Cd) on the defense responses of the Pacific oyster Crassostrea gigas to Listonella anguillarum challenge.The activities of several important defensive enzymes,including superoxide dismutase(SOD),glutathione peroxidase(GPx),acid phosphatase(ACP),Na+,K+-ATPase in gills and hepatopancreas,and phenoloxidase-like(POL) enzyme in hemolymph were assayed.In addition,the expression levels of several genes,including heat shock protein 90(HSP90),metallothionein(MT),and bactericidal/permeability increasing(BPI) protein were quantified by fluorescent quantitative PCR.The enzyme activities of SOD,ACP,POL,and GPx in hepatopancreas,and the expression of HSP90 were down-regulated,whereas GPx activity in the gill,Na+,K+-ATPase activities in both tissues,and MT expression was increased in Cdexposed oysters post L.anguillarum challenge.However,BPI expression was not significantly altered by co-stress of L.anguillarum infection and cadmium exposure.Our results suggest that cadmium exposure alters the oysters’ immune responses and energy metabolism following vibrio infection.  相似文献   

15.
In this paper, fluxes of momentum and sensible heat are discussed with the data collected by tethered balloon sounding system over the Philippine Sea during the cruise of the R/VSCIENCE 1 from September through October in 1987. These fluxes were calculated using the semempirical flux-profile relationships of Monin-Obkhov similarity theory with observed data. The friction velocityU. was determined by the observed data's least-square fit with the similarity formulae under stable, neutral and unstable conditions. The roughnessZ 0 was determined byZ 0=a1 (U*/g), then substituted into the similarity formulae to computeU * again. The final values ofU * andZ 0 could be determined through this iteration. The flux temperatureθ * was calculated from the temperature profile withZ 0 determined above. Finally the fluxes of momentum and sensible heat, and the drag coefficientC D were obtained by computation withU * θ * and the wind speed (U 10) at 10 meters above the sea surface. Contribution No. 2220 from the Institute of Oceanology, Chinese Academy of Sciences. This project was supported by the National Natural Science, Foundation of China (No. 049176255).  相似文献   

16.
Hyper-spectrum models for monitoring water quality in Dianshan Lake,China   总被引：1，自引：0，他引：1  
The correlation between water quality parameters and hyper-spectral reflectance is studied with models established for each parameter and applied in Dianshan Lake, in the upstream of the Huangpu River running through Shanghai, China. Models are for dissolved oxygen (DO in mg/L): R720/R680 = 20.362×(R720/R680)2-31.438×(R720/R680)+19.156; for turbidity (NTU): R*714.5 = 206.07× (R*714.5)2-582.5×R*714.5 + 423.24; and for total phosphorus (TP in mg/L): R*509.5 = 16.661× (R*509.5)2-32.646×R*509.5+16.116. The R2 values are 0.770 8, 0.660 4 and 0.738 7, respectively, showing strong positive relationships. The models were then applied to assess water quality of different times. Results are quite satisfactory for some samples.  相似文献   

17.
Extracellular products (ECP) produced byVibrio anguillarum strain M3 originally isolated from diseased flounder (Paralichthys olivaceus) were prepared. ECP of M3 showed gelatinase, casinase, amylase and haemolytic activity on agarose plates. High protease activity against azocasin was detected. Bacterium M2 showed highest growth and protease activity at 25°C. The protease present in ECP showed maximal activity at pH 8 and 55°C; was completely inactivated by application of 80°C heat for 30 min; was completely inhibited by EDTA and HgCl2, and was partially inhibited by PMSF, SDS, MnCl2 and iodoacetic acid; but not inhibited by CaCl2 and MgCl2. The ECP was toxic to flounder fish at LD50 values of 3.1 μg protein/g body weight. The addition of HgCl2 and application of heat at 50°C decreased the lethal toxicity of ECP. When heated at 100°C, ECP lethality to flounder was completely inhibited. After intramuscular injection of ECP into flounder, it showed evident histopathological changes including necrosis of muscle, extensive deposition of haemosiderin in the spleen, dilated blood vessels congested with numerious lymphocytes in the liver. These results showed that ECP protease was a lethal factor produced by the bacteriumV. anguillarum M3. Contribution No. 4213 from the Institute of Oceanology, Chinese Academy of Sciences. Funded by projects under the Major State Basic Research Development Program, Grant G1999012003.  相似文献   

18.
1 INTRODUCTIONIt is well known that wind-generated wavestorm surge and atmospheric circulation as impor-tant ocean weather events are closely related tomomentum exchange between sea and atmosphere(Oncley and Dudhia, 1995). For this reason, in thepast 20 years, attention has been directed towardshow to better describe momentum exchange throughsea surface. In general, to precisely determine theexchange, improvement should be made in the as-pects of reasonably depicting the roughness de-pend…  相似文献   

19.
The diffuse attenuation coefficient for downwelling irradiance （Kd（λ）） is an important parameter for ocean studies. Based on the optical profile data measured during three cruises in the northern South China Sea in autumn from 2003 to 2005, variations in the Kd（λ） spectra were analyzed. The variability of Kd（λ） shows much distinct features in both magnitude and spectra pattern, it is much higher in coastal waters than that of open oceanic waters; and the blue-to-green （443/555） ratio of Kd（λ） tends to increase with chlorophyll a concentration （[Chl-a]） from open ocean to coastal waters. These characteristics can be explained most by the increase of aw＋p（443）/aw＋p（555） with [Chl-a]. In short waveband, the relation between Kd（λ）-Kw（λ） and [Chl-a] can be well described by a power law function, indicating the large contribution of phytoplankton to the variations in Kd（λ）. As for the spectral model of the diffuse attenuation coefficient, there are good linear relationships between Kd（490） and Kd（λ） in other wavelengths with own slope and intercept of a linear functions in the spectral range 412-555 nm. Kd（490） is well correlated with the spectral ratio of remote sensing reflectance; and should enough measurement data are given, this empirical algorithm would be used in the Kd（λ） retrieval from ocean color satellite data. The variation in Kd（λ） provides much useful information for us to study the bio-optical property in the northern South China Sea.  相似文献   

20.
Wang  Huanshan  Wang  Teng  Li  Wenjing  Liu  Huanzhang 《中国海洋湖沼学报》2019,37(3):1042-1050
Megalobrama pellegrini is a cyprinid fish endemic to upper reaches of the Changjiang(Yangtze) River, China, which is also an important economic species in the local area. In recent years,resources of this species have decreased sharply and its conservation has drawn great attention. In the present study, we collected 120 individuals from the Longxi River, a tributary isolated from the main channel of the Changjiang River, where M. pellegrini is still relatively abundant. Using two different molecular markers, mitochondrial cytochrome b(cyt b) gene, and nuclear microsatellite(simple sequence repeat,SSR), we analyzed the genetic diversity of this isolated population. The results show that sequence genetic diversity was low(H_d:0.290 and P_i: 0.000 77 for cyt b gene),while the SSR genetic diversity was high(H_o:0.824 4±0.147 2,H_e: 0.823 510.145 1). Analysis indicated that this population had experienced a bottleneck,with inbreeding and small effective population size(around 50). Based on SSR data, relatedness analyzing revealed that the 120 samples were grouped into 10 completely independent clusters. It was inferred that the mating system of M. pellegrini was polygamy. We suggested that the low genetic diversity could be induced by the overfishing and inbreeding depression. Therefore, we suggested that the urgent conservation measures should be taken to control the overfishing and give better conditions for the fish to grow and spawn, then to restore population size.  相似文献   

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