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Flavobacterium columnare causes columnaris disease in freshwater fish. In the present study, the antigenic regions of five outer membrane proteins (OMPs), including zinc metalloprotease, prolyl oligopeptidase, thermolysin, collagenase and chondroitin AC lyase, were bioinformatically analyzed, fused together, and then expressed as a recombinant fusion protein in Escherichia coli. The expressed protein of 95.6 kDa, as estimated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis, was consistent with the molecular weight deduced from the amino acid sequence. The purified recombinant protein was used to vaccinate the grass carp, Ctenopharyngodon idella. Following vaccination of the fish their IgM antibody levels were examined, as was the expression of IgM, IgD and IgZ immunoglobulin genes and other genes such as MHC Iα and MHC IIβ, which are also involved in adaptive immunity. Interleukin genes (IL), including IL-1β, IL-8 and IL-10, and type I and type II interferon (IFN) genes were also examined. At 3 and 4 weeks post-vaccination (wpv), significant increases in IgM antibody levels were observed in the fish vaccinated with the recombinant fusion protein, and an increase in the expression levels of IgM, IgD and IgZ genes was also detected following the vaccinations, thus indicating that an adaptive immune response was induced by the vaccinations. Early increases in the expression levels of IL and IFN genes were also observed in the vaccinated fish. At four wpv, the fish were challenged with F. columnare, and the vaccinated fish showed a good level of protection against this pathogen, with 39% relative percent survival (RPS) compared with the control group. It can be concluded, therefore, that the five OMPs, in the form of a recombinant fusion protein vaccine, induced an immune response in fish and protection against F. columnare.  相似文献   

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Acaudina leucoprocta is an edible sea cucumber of economic interest that is widely distributed in China. Little information is available concerning the molecular genetics of this species although such knowledge would contribute to a better understanding of the optimal conditions for its aquaculture and its mechanisms of defense against disease. Therefore, we constructed a cDNA library and, based on bioinformatics analysis of the sequences, the functions of 75% of the cDNAs were identified, including those involved in cell structure, energy metabolism, mitochondrial function, and signal transduction pathways. Approximately 25% of genes in the library were unmatched. The gene for A. leucoprocta ferritin was also cloned. The predicted amino-acid sequence of ferritin displayed significant homology with other sea-cucumber counterparts but indicated that it was a new member of the ferritin family. Semiquantitative real-time RT-PCR indicated the highest levels of ferritin mRNA expression in the intestine. A polyclonal antibody of ferritin was also produced. These data provide a set of molecular tools essential for further studies of the functions of ferritin protein in A. leucoprocta.  相似文献   

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Selection of net with a suitable mesh size is a key concern in the quantitative assessment of zooplankton, which is crucial to understand pelagic ecosystem processes. This study compared the copepod collecting efficiency of three commonly used plankton nets, namely, the China standard coarse net(505 μm mesh), the China standard fine net(77 μm), and the WP-2 net(200 μm). The experiment was performed at six stations in the Bohai Sea during the autumn of 2012. The coarse net substantially under-sampled small individuals(body widths 672 μm) and led to the lowest species number in each tow, whereas the fine net collected all small copepod species but failed to collect rare species. The WP-2 net appeared to be a compromise of the two other nets, collecting both small copepods and rare species. The abundance of copepods collected by the coarse net(126.4 ± 86.5 ind m~(-3)) was one to two orders of magnitude lower than that by the WP-2 net(5802.4 ± 2595.4 ind m~(-3)), and the value of the fine net(11117.0 ± 4563.41 ind m~(-3)) was nearly twice that of the WP-2 net. The abundance of large copepods(i.e., adult Calanus sinicus) in the three nets showed no significant differences, but the abundance of small copepods declined with decreasing mesh size. The difference in abundance resulted from the under-sampling of small copepods with body widths 672 μm and 266 μm by the coarse and WP-2 nets, respectively.  相似文献   

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Members of the DnaJ family are proteins that play a pivotal role in various cellular processes, such as protein folding, protein transport and cellular responses to stress. In the present study, we identified and characterized the full-length DnaJ cDNA sequence from expressed sequence tags of Pyropia yezoensis (PyDnaJ) via rapid identification of cDNA ends. This cDNA encoded a protein of 429 amino acids, which shared high sequence similarity with other identified DnaJ proteins, such as a heat shock protein 40/DnaJ from Pyropia haitanensis. The relative mRNA expression level of PyDnaJ was investigated using real-time PCR to determine its specific expression during the algal life cycle and during desiccation. The relative mRNA expression level in sporophytes was higher than that in gametophytes and significantly increased during the whole desiccation process. These results indicate that PyDnaJ is an authentic member of the DnaJ family in plants and red algae and might play a pivotal role in mitigating damage to P. yezoensis during desiccation.  相似文献   

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Dominant species of zooplankton community vary with latitude. Though China possesses a vast coastal area in northwestern Pacific, studies on the latitudinal dominant species gradient are rare. We collected zooplankton samples from Haizhou Bay(34.56?–35.19?N, 119.51?–120.30?E), Yueqing Bay(28.14?–28.38?N, 121.10?–121.21?E) and Dongshan Bay(23.65?–23.90?N, 117.45?–117.60?E) in May 2012 and May 2013 to preliminarily characterize the latitudinal dominant species distribution. All the samples were collected vertically using a 0.505 mm mesh plankton net with 0.8 m in mouth diameter from bottom to surface. Calanus sinicus, Aidanosagitta crassa, Labidocera euchaeta, Zonosagitta nagae, Acartia pacifica and Paracalanus parvus were found to be dominant. C. sinicus was the most dominant species and the unique one occurred in all three bays. With latitude decreasing, both the abundance and proportion of C. sinicus declined sharply. Cluster analysis showed that the 6 dominant species could be divided into 3 groups, based on their occurrences in the three bays. Our results suggested that the distribution of dominant species along the coast of China has a significant latitudinal gradient. C. sinicus which widely distributes in the coastal water of the northwestern Pacific can well adapt to the temperature at different latitudes. The high abundance in Haizhou Bay indicated that C. sinicus was an exemplary warm-temperate species, and more commonly occurs in the north of China seas. The ecological characteristics of dominant species change from warm-temperate type in high-latitudinal bays to warm water type in low-latitudinal bays.  相似文献   

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Water vapor in the earth′s upper atmosphere plays a crucial role in the radiative balance, hydrological process, and climate change. Based on the latest moderate-resolution imaging spectroradiometer(MODIS) data, this study probes the spatio-temporal variations of global water vapor content in the past decade. It is found that overall the global water vapor content declined from 2003 to 2012(slope b = –0.0149, R = 0.893, P = 0.0005). The decreasing trend over the ocean surface(b = –0.0170, R = 0.908, P = 0.0003) is more explicit than that over terrestrial surface(b = –0.0100, R = 0.782, P = 0.0070), more significant over the Northern Hemisphere(b = –0.0175, R = 0.923, P = 0.0001) than that over the Southern Hemisphere(b = –0.0123, R = 0.826, P = 0.0030). In addition, the analytical results indicate that water vapor content are decreasing obviously between latitude of 36°N and 36°S(b = 0.0224, R = 0.892, P = 0.0005), especially between latitude of 0°N and 36°N(b = 0.0263, R = 0.931, P = 0.0001), while the water vapor concentrations are increasing slightly in the Arctic regions(b = 0.0028, R = 0.612, P = 0.0590). The decreasing and spatial variation of water vapor content regulates the effects of carbon dioxide which is the main reason of the trend in global surface temperatures becoming nearly flat since the late 1990 s. The spatio-temporal variations of water vapor content also affect the growth and spatial distribution of global vegetation which also regulates the global surface temperature change, and the climate change is mainly caused by the earth's orbit position in the solar and galaxy system. A big data model based on gravitational-magmatic change with the solar or the galactic system is proposed to be built for analyzing how the earth's orbit position in the solar and galaxy system affects spatio-temporal variations of global water vapor content, vegetation and temperature at large spatio-temporal scale. This comprehensive examination of water vapor changes promises a holistic understanding of the global climate change and potential underlying mechanisms.  相似文献   

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There is considerable concern that isoflavones, such as genistein in fish feed composed of soybean protein, aff ects somatic growth in fish. Our previous works demonstrated that 30 and 300 μg/g dietary genistein had no significant eff ect on growth performance in Nile tilapia (Oreochromis niloticus), but the higher level of genistein (3 000 μg/g) significantly depressed growth. This study was conducted to further examine the eff ects of dietary genistein on the endocrine disruption on growth hormone/insulin-like growth factor-I (GH/IGF-I) axis in Nile tilapia (O. niloticus). Juvenile fish were fed by hand twice daily to satiation with one of four isonitrogenous and isoenergetic diets, each containing either 0, 30, 300 or 3 000 μg/g genistein. Following an 8-week feeding period, plasma GH and IGF-I levels were investigated by radioimmunoassay and gene expression levels of gh, ghrelin, gnrhs, ghr, npy, npyrs, pacap, ghrs, igf-I, igf-Ir, and igfbp3 were examined by real-time PCR. The results show that no significant change in plasma GH and IGF-I levels in fish fed with diets containing 30 μg/g and 300 μg/g genistein. mRNA expression of genes along the GH/IGF-I axis remained unaff ected, except for igf-Ir, which was stimulated by the 300 μg/g genistein diet. While in fish fed the 3 000 μg/g genistein diet, the plasma GH and IGF-I levels decreased, and mRNA expression of gh, ghr2, npyr1, igf-I, and igf-Ir were also significantly depressed. In contrast, npy and igfbp3 mRNA expression were enhanced. This study provides convincing evidence for growth impediment by genistein by disturbing the GH/IGF-I axis in Nile tilapia O. niloticus.  相似文献   

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An interannual study on zooplankton abundance, biomass, and species composition was carried out during different seasons in two local coastal water types off Gopalpur, north-western Bay of Bengal. Although, Type-1 was observed with higher zooplankton abundance in comparison to Type-2, pattern of variation followed similar seasonal trends in both water types during individual years. Well pronounced seasonality was observed in zooplankton distribution. Zooplankton community was composed of 217 holoplankton and 22 meroplankton. The holoplankton community was predominated by copepod in terms of species diversity and abundance followed by hydrozoa, tintinnida, malacostraca, gastropoda, chaetognatha and chordata. The meroplankton were represented by larval forms viz. bivalve veliger, brachyuran zoea larvae, caridean larvae, copepod nauplii, fish egg and gastropod veliger. Dominance of copepod species viz. Acrocalanus longicornis, Paracalanus aculeatus and Paracalanus parvus were observed frequently in both water types. The species richness was higher in Type-1 in comparison to Type-2 during both the years. Salinity regimes and availability of phytoplankton prey influenced the distribution and species composition of zooplankton assemblage.  相似文献   

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The pattern recognition proteins (PRPs) play a major role in immune response of crustacean to resist pathogens. In the present study, as one of PRPs, lipopolysaccharide and β-1, 3-glucan binding protein (LGBP) gene in the ridge tail white prawn (Exopalaemon carinicauda) (EcLGBP) was isolated. The full-length cDNA of EcLGBP was 1338 bp, encoding a polypeptide of 366 amino acid residules. The deduced amino acid sequence of EcLGBP shared high similarities with LGBP and BGBP from other crustaceans. Some conservative domains were predicted in EcLGBP sequence. EcLGBP constitutively expressed in most tissues at different levels, and the highest expression was observed in hepatopancreas. With infection time, the cumulative mortality increased gradually followed by the proliferation of Vibrio parahaemolyticus and white spot syndrome virus (WSSV). The expression of EcLGBP in response to V. parahaemolyticus infection was up-regulated in hemocytes and hepatopancreas, and the up-regulation in hepatopancreas was earlier than that in hemocytes. EcLGBP expression after WSSV infection increased at 3 h, then significantly decreased in both hemocytes and hepatopancreas. The results indicated that EcLGBP was involved in the immune defense against bacterial and viral infections.  相似文献   

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The RNA helicase Vasa is an important regulator of primordial germ cell development. Its function in mature fish, especially the hormone-related differences in maturing male fish has seldom been documented. In this study, a full length cDNA sequence of the vasa gene was cloned from Japanese sea bass, Lateolabrax japonicas, and it was named jsb-vasa. Homology analysis showed that jsb-vasa was closely related to its teleost homologs. The spatial distribution of jsb-vasa indicated that it was only highly expressed in testis, showing its germ cell-specific expression pattern. During the testicular development cycle, jsb-vasa was highly expressed during early period of spermatogenesis, and reduced when spermatogenesis advanced. In addition, the jsb-vasa gene expression was significantly inhibited at 6 h, 12 h and 24 h after injecting hCG (human chorionic gonadotropin) and GnRHa (Gonadotropin-releasing hormone analogue), indicating that jsb-vasa gene may play an important role in spermatogenesis of Japanese sea bass, and be under the regulation of external sex hormones.  相似文献   

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The effects of different phosphorus (P) concentrations (0.36, 3.6, and 36 μmol/L corresponding to low-, middle-, and high-P concentration groups, respectively) and nitrogen (N)/P ratios on the growth and photosynthetic characteristics of Skeletonema costatum and Prorocentrum donghaiense were studied. For both species, the high-P (HP) concentration group showed the greatest algal density and highest specific growth rate. Changes in the maximum efficiency of photosystem II (F v /F m ) were monitored under the various P and N/P conditions. The largest decrease in F v /F m was in the low-P (LP) group in S. costatum and in the HP group in P. donghaiense. There were high rapid light curves and photochemical quantum yields (Φ PSII) for S. costatum in the HP group, while the actual photosynthetic capacity was higher in P. donghaiense than in S. costatum in the MP group. Under eutrophic but relatively P-restricted conditions, P. donghaiense had higher photosynthetic activity and potential, which could cause this dinoflagellate to increasingly dominate the phytoplankton community in these conditions. Under the same P concentration and N/P ratio, P. donghaiense had a larger relative maximum rate of electron transport and higher Φ PS II values than those of S. costatum. These differences between P. donghaiense and S. costatum may explain the interaction and succession patterns of these two species in the Changjiang (Yangtze) River estuary from a photosynthesis perspective.  相似文献   

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Soil respiration (SR) is the second-largest flux in ecosystem carbon cycling. Due to the large spatio-temporal variability of environmental factors, SR varied among different vegetation types, thereby impeding accurate estimation of CO2 emissions via SR. However, studies on spatio-temporal variation of SR are still scarce for semi-arid regions of North China. In this study, we conducted 12-month SR measurements in six land-use types, including two secondary forests (Populus tomentosa (PT) and Robinia pseudoacacia (RP)), three artificial plantations (Armeniaca sibirica (AS), Punica granatum (PG) and Ziziphus jujuba (ZJ)) and one natural grassland (GR), to quantify spatio-temporal variation of SR and distinguish its controlling factors. Results indicated that SR exhibited distinct seasonal patterns for the six sites. Soil respiration peaked in August 2012 and bottomed in April 2013. The temporal coefficient of variation (CV) of SR for the six sites ranged from 76.98% to 94.08%, while the spatial CV of SR ranged from 20.28% to 72.97% across the 12-month measurement. Soil temperature and soil moisture were the major controlling factors of temporal variation of SR in the six sites, while spatial variation in SR was mainly caused by the differences in soil total nitrogen (STN), soil organic carbon (SOC), net photosynthesis rate, and fine root biomass. Our results show that the annual average SR and Q10 (temperature sensitivity of soil respiration) values tended to decrease from secondary forests and grassland to plantations, indicating that the conversion of natural ecosystems to man-made ecosystems may reduce CO2 emissions and SR temperature sensitivity. Due to the high spatio-temporal variation of SR in our study area, care should be taken when converting secondary forests and grassland to plantations from the point view of accurately quantifying CO2 emissions via SR at regional scales.  相似文献   

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