Zhejiang Institute of Freshwater Fisheries

Huzhou, China

Zhejiang Institute of Freshwater Fisheries

Huzhou, China

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Sun S.,East China Normal University | Sun S.,Zhejiang Normal University | Ye J.,Zhejiang Normal University | Chen J.,Zhejiang Institute of Freshwater Fisheries | And 2 more authors.
Aquaculture Nutrition | Year: 2011

An 8-week experiment on fingerling black carp Mylopharyngodon piceus was conducted to evaluate the effects of dietary fish oil (FO) supplement on growth, fatty acid composition and non-specific immunity responses. Five triplicate fingerling groups (initial weight=2.72±0.35g) were fed isoenergetic and isonitrogenous diets in which the dietary FO was replaced with rapeseed oil (RO) in graded increments of 25% (0-100%). No significant effects were observed on specific growth rates, survival rates and feed conversion ratios, but there were significant differences in whole body moisture and liver lipid contents (P<0.05), and the 100% RO replacement diet significantly enhanced hepatosomatic indexes compared to control group (P< 0.05). Other approximate whole body constituents, viscerasomatic ratios and condition factors were not influenced by dietary oil treatments. Fatty acid composition of muscle and liver was influenced by dietary fatty acid input, α-linoleic acid and γ-linolenic acid were significantly increased with increasing RO, but eicosapentaenoic acid, docosahexaenoic acid and the n-3/n-6 ratio were significantly reduced (P<0.05). Alternative complement pathway, lysozyme and superoxide dismutase activities were not significantly influenced. These results indicate that black carp fed diets with FO supplement had similar growth and non-specific immunity to the fish fed diet with RO. © 2010 Blackwell Publishing Ltd.


Yao J.-Y.,Zhejiang Institute of Freshwater Fisheries | Yin W.-L.,Zhejiang Institute of Freshwater Fisheries | Li X.-C.,CAS East China Sea Fisheries Research Institute | Li G.,National Fisheries Technology Extension Center | And 3 more authors.
Parasitology Research | Year: 2014

The ciliate Ichthyophthirius multifiliis is one of the most pathogenic parasites of fish maintained in captivity. In this study, effects of bacterial extracellular products of Streptomyces griseus SDX-4 against I. multifiliis were determined. The fermentation liquor of S. griseus was extracted successively in a separating funnel with petroleum ether, ethyl acetate, and n-butanol. In vitro assays revealed that the n-butanol extracts (NBu-E) and ethyl acetate extracts (Eto-E) of S. griseus were observed to be more effective against theronts than the other extracts with median effective concentration (EC 50) values of 0.86 and 12.5 mg L-1, respectively, and significantly reduced the survival of the tomonts and the total number of theronts released by the tomonts (P<0.05). All encysted tomonts were killed when the concentration of NBu-E was 30.0 mg L-1. Results of in vivo test demonstrated that the number of I. multifiliis trophonts on the grass carp treated with NBu-E was markedly lower compared to the control group at 11 days after exposed to theronts (P<0.05). In the control group, 100 % mortality was observed owing to heavy I. multifiliis infection at 11 days after the exposure. On the other hand, only 9.5 % mortality owing to parasite infection was recorded in the groups treated with the NBu-E (30 mg L-1). The median lethal dose (LD50) of NBu-E for grass carp was 152.4 mg L -1. Our results indicate that n-butanol extract of S. griseus will be useful in aquaculture for controlling I. multifiliis infections. © 2014 Springer-Verlag.


Long Y.,CAS Wuhan Institute of Hydrobiology | Li Q.,CAS Wuhan Institute of Hydrobiology | Zhong S.,Wuhan University | Wang Y.,Zhejiang Institute of Freshwater Fisheries | Cui Z.,CAS Wuhan Institute of Hydrobiology
Comparative Biochemistry and Physiology - C Toxicology and Pharmacology | Year: 2011

Multidrug-resistance associated protein 2 (MRP2/ABCC2) plays crucial roles in bile formation and detoxification by transporting a wide variety of endogenous compounds and xenobiotics, but its functions in zebrafish (Danio rerio) remain to be characterized. In this study, we obtained the full-length cDNA of zebrafish abcc2, analyzed its expression in developing embryos and adult tissues, investigated its transcriptional response to heavy metals, and evaluated its roles in efflux of heavy metals including cadmium, mercury and lead. Zebrafish abcc2 gene is located on chromosome 13 and composed of 32 exons. The deduced polypeptide of zebrafish ABCC2 consists of 1567 amino acids and possesses most of functional domains and critical residues defined in human ABCC2. Zebrafish abcc2 gene is not maternally expressed and its earliest expression was detected in embryos at 72 hpf. In larval zebrafish, abcc2 gene was found to be exclusively expressed in liver, intestine and pronephric tubules. In adult zebrafish, the highest expression of abcc2 gene was found in intestine followed by those in liver and kidney, while relative low expression was detected in brain and muscle. Expression of abcc2 in excretory organs including kidney, liver and intestine of zebrafish larvae was induced by exposure to 0.5 μM mercury or 5 μM lead. Moreover, exposure to 0.125-1 μM of mercury or lead also significantly induced abcc2 expression in these excretory organs of adult zebrafish. Furthermore, overexpression of zebrafish ABCC2 in ZF4 cells and zebrafish embryos decreased the cellular accumulation of heavy metals including cadmium, mercury and lead as determined by MRE (metal responsive element)- or EPRE (electrophile response element)-driven luciferase reporters and atomic absorption spectrometry. These results suggest that zebrafish ABCC2/MRP2 is capable of effluxing heavy metals from cells and may play important roles in the detoxification of toxic metals. © 2011 Elsevier Inc.


Liu W.,Ningbo University | Liu W.,Zhejiang Institute of Freshwater Fisheries | Qian D.,Zhejiang Institute of Freshwater Fisheries | Yan X.,Ningbo University
Aquaculture | Year: 2011

To better understand the molecular pathogenesis of white spot syndrome virus (WSSV) in mud crab Scylla serrata, the two-dimensional electrophoresis (2-DE) protein expression profiles of the hemolymph from the unchallenged and WSSV challenged S. serrata were compared. Finally, 22 differentially expressed protein spots (12 spots with up-regulated protein expression levels and 10 spots with down-regulated levels) were successfully identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) analysis. The up-regulated proteins included hemocyanin, alpha-2 macroglobulin (A2M), heat shock protein 70 (Hsp70), cytosolic manganese superoxide dismutase (MnSOD), catalase (CAT), cryptocyanin, glutamine synthetase, arginine kinase and ribosomal protein S28. The down-regulated proteins included heat shock protein 90 (Hsp90), cryptocyanin 1, prophenoloxidase (proPO), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), actin, tropomyosin, synaptosomal-associated protein (SNAP) and transposase. The differentially expressed proteins participate in various important functions such as host defense responses, glycolytic and energy metabolism. The data may help to better understanding of the mechanism of molecular responses of S. serrata to WSSV infection. © 2011 Elsevier B.V.


Liu W.,Ningbo University | Liu W.,Zhejiang Institute of Freshwater Fisheries | Qian D.,Zhejiang Institute of Freshwater Fisheries | Yan X.J.,Ningbo University
Journal of Fish Diseases | Year: 2011

Mud crab, Scylla serrata (Forskal), is the most commercially important marine crab species in China. In recent years, serious diseases have occurred in major mud crab culture regions in SE China. PCR detection of white spot syndrome virus (WSSV) in diseased mud crabs collected from Zhejiang Province during 2006-2008 showed a prevalence of 34.82%. To study the pathogenicity of WSSV to mud crab, healthy mud crabs were injected intramuscularly with serial 10-fold dilutions of a WSSV inoculum. The cumulative mortalities in groups challenged with 10-1, 10-2, 10-3 and 10-4 dilutions were 100%, 100%, 66.7% and 38.9% at 10 days post-injection, respectively. All moribund and dead mud crabs except the control group were positive for WSSV by PCR. Based on the viral load of the WSSV inoculum by quantitative real-time PCR, the median lethal dose (LD50) of WSSV in S. serrata was calculated as 1.10 × 106 virus copies/crab, or 7.34 × 103 virus copies g-1 crab weight. The phenoloxidase, peroxidase and superoxide dismutase activities in haemolymph of WSSV-infected moribund crabs, were significantly lower than the control group, whereas alkaline phosphatase, glutamate-pyruvate transaminase and glutamic-oxaloacetic transaminase were higher than in the control group. WSSV was mainly distributed in gills, subcuticular epithelia, heart, intestine and stomach as shown by immunohistochemical analysis with Mabs against WSSV. The epithelial cells of infected gill showed hypertrophied nuclei with basophilic inclusions. Numerous bacilliform virus particles were observed in nuclei of infected gill cells by transmission electron microscopy. It is concluded that WSSV is a major pathogen of mud crab with high pathogenicity. © 2011 Blackwell Publishing Ltd.


Liu J.Y.,CAS Wuhan Institute of Hydrobiology | Liu J.Y.,University of Chinese Academy of Sciences | Li A.H.,CAS Wuhan Institute of Hydrobiology | Zhou D.R.,Zhejiang Institute of Freshwater Fisheries | And 2 more authors.
Aquaculture Research | Year: 2010

Yellow catfish Pelteobagrus fulvidraco (Richardson) is a commercially important fish generally distributed in Southeast Asian countries. The well-known aetiological agent of enteric septicaemia of catfish, Edwardsiella ictaluri, was isolated from diseased yellow catfish P. fulvidraco (Richardson) reared at two commercial fisheries in China. The economic losses due to the high mortalities (about 50%) caused by this bacterium have been increasing annually. The affected fish presented two different, typical symptoms: pale gills, slight exophthalmia and a 'hole in the head', and haemorrhage on the opercula, in the skin under the jaw, creating a 'hole under the jaw'. These diseases were found frequently in cultured yellow catfish throughout China. The isolates from both outbreaks were all Gram negative, facultatively anaerobic and short rod. Morphological and biochemical tests and phylogenetic analysis based on the 16S rDNA sequences all strongly indicated that these yellow catfish isolates were highly identical to the known E. ictaluri. In addition, the isolates possessed the typical plasmid profile of E. ictaluri. Experimental infection assays were conducted and pathogenicity (by an intraperitoneal injection) was demonstrated in yellow catfish and channel catfish Ictalurus punctatus. The results showed that yellow catfish isolates were quite conservative phenotypically and genetically, and were able to cause two different, typical symptoms in this fish under unknown conditions and mechanism. © 2010 The Authors. Aquaculture Research © 2010 Blackwell Publishing Ltd.


Sun X.,Zhejiang University | Ruan R.,Zhejiang University | Lin L.,Zhejiang Institute of Freshwater Fisheries | Zhu C.,Zhejiang University | And 4 more authors.
FEMS Microbiology Letters | Year: 2013

Penicillium digitatum, causing citrus green mold, is one of the most devastating pathogenic fungi for postharvest fruits. The disease control is becoming less efficient because of the dispersal of fungicide-resistant strains. However, genome-scale analyses of its resistance mechanism are scarce. In this work, we sequenced the whole genome of the R1 genotype strain Pd01-ZJU and investigated the genes and DNA elements highly associated with drug resistance. Variation in DNA elements related to drug resistance between P. digitatum strains was revealed in both copy number and chromosomal location, indicating that their recent and frequent translocation might have contributed to environmental adaptation. In addition, ABC transporter proteins in Pd01-ZJU were characterized, and the roles of typical subfamilies (ABCG, ABCC, and ABCB) in imazalil resistance were explored using real-time PCR. Seven ABC proteins, including the previously characterized PMR1 and PMR5, were induced by imazalil, which suggests a role in drug resistance. In summary, this work presents genome information of the R1 genotype P. digitatum and systematically investigates DNA elements and ABC proteins associated with imazalil resistance for the first time, which would be indicative for studying resistant mechanisms in other pathogenic fungi. © 2013 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.


Yang X.,Zhejiang University | Sun J.-Y.,Zhejiang University | Guo J.-L.,Zhejiang Institute of Freshwater Fisheries | Weng X.-Y.,Zhejiang University
Journal of the Science of Food and Agriculture | Year: 2012

Background: Cottonseed meal, an important source of feed raw materials, has limited use in the feed industry because of the presence of the highly toxic gossypol. The aim of the current work was to isolate the gossypol-degrading fungus from a soil microcosm and investigate the proteins involved in gossypol degradation. Results: A fungal strain, AN-1, that uses gossypol as its sole carbon source was isolated and identified as Aspergillus niger. A large number of intracellular proteins were detected using sodium dodecyl sulfate-polyacrylamide gel electrophoresis, but no significant difference was observed between the glucose-containing and gossypol-containing mycelium extracts. Two-dimensional gel electrophoresis results showed that the protein spots were concentrated in the 25.0-66.2 kDa range and distributed in different pI gradients. PDQuest software showed that 51 protein spots in the gels were differentially expressed. Of these, 20 differential protein spots, including six special spots expressed in gossypol, were analyzed using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Conclusion: The fungus AN-1 biodegraded gossypol and the proteomic analysis results indicate that some proteins were involved in the gossypol biodegradation during fungus survival, using gossypol as its sole carbon source. © 2011 Society of Chemical Industry.


Yang X.,Zhejiang University | Guo J.L.,Zhejiang Institute of Freshwater Fisheries | Ye J.Y.,Zhejiang University | Zhang Y.X.,Zhejiang University | Wang W.,Zhejiang University
Fish and Shellfish Immunology | Year: 2015

The present study investigated the effect of Ficus carica polysaccharide (FCP), isolated from the fruit of F. carica L., at 0%, 0.1%, 0.5% and 1.0% doses supplementation with feed on genes Interleukin 1-β (IL-1β), Tumor Necrosis Factor α (TNF-α) and heat shock protein 70 (HSP70) gene expression in blood, humoral innate immune parameters and resistant to Flavobacterium columnare of grass carp at weeks 1, 2 and 3. The results revealed that administration of FCP significantly (P<0.05) up regulated IL-1β and TNF-α gene expression. HSP70 gene expression was significantly (P<0.05) lower in FCP-fed fish at the end of trial. The serum total protein, albumin and globulin did not significantly increased in any diet on the first week whereas it was significantly enhanced in 0.5% and 1.0% supplementation diets on weeks 2 and 3 when compared to control. The serum complement C3 was significantly (P<0.05) increased on weeks 1 and 2 when compared to control, however, no significant difference was found in this activity after 3 weeks of treatment. All diets significantly enhanced the serum lysozyme activity, bactericidal activity from weeks 1-2 as compared to control. Grass carp fed with FCP showed remarkably higher resistance against F.columnare (60% survival) compared to the control group (30% survival). These results confirm that FCP can up regulate immune related genes expression, stimulates immune response that per se enhances disease resistance in grass carp. © 2014 Elsevier Ltd.


Long Y.,CAS Wuhan Institute of Hydrobiology | Li Q.,CAS Wuhan Institute of Hydrobiology | Wang Y.,Zhejiang Institute of Freshwater Fisheries | Cui Z.,CAS Wuhan Institute of Hydrobiology
Comparative Biochemistry and Physiology - C Toxicology and Pharmacology | Year: 2011

Acquired resistance of mammalian cells to heavy metals is closely relevant to enhanced expression of several multidrug resistance-associated proteins (MRP), but it remains unclear whether MRP proteins confer resistance to heavy metals in zebrafish. In this study, we obtained zebrafish (Danio rerio) fibroblast-like ZF4 cells with resistance to toxic heavy metals after chronic cadmium exposure and selection for 6 months. These cadmium-resistant cells (ZF4-Cd) were maintained in 5 μM cadmium and displayed cross-resistance to cadmium, mercury, arsenite and arsenate. ZF4-Cd cells remained the resistance to heavy metals after protracted culture in cadmium-free medium. In comparison with ZF4-WT cells, ZF4-Cd cells exhibited accelerated rate of cadmium excretion, enhanced activity of MRP-like transport, elevated expression of abcc2, abcc4 and mt2 genes, and increased content of cellular GSH. Inhibition of MRP-like transport activity, GSH biosynthesis and GST activity significantly attenuated the resistance of ZF4-Cd cells to heavy metals. The results indicate that some of MRP transporters are involved in the efflux of heavy metals conjugated with cellular GSH and thus play crucial roles in heavy metal detoxification of zebrafish cells. © 2010 Elsevier Inc.

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