International Journal of Aquaculture

ConferenceⅠConference位于名称中间，有介词。

Group 1The 2009 International Conference on Information and Communications SecurityThe 2010 IEEE International Conference on Networking, Sensing and Control (ICNSC 2010)ICPR 2010 - The 20th International Conference on Pattern Recognition International Conference on Spoken Language Processing (ICSLP)(ICASSP ’98)ANZALS Leisure & Recreation Conference 2008: Leisure is the keyThe Eleventh IEEE International Conference on Computer Vision (ICCV 2007) The 18th International Conference on Pattern Recognition (ICPR 06)The 7th International Conference on Functional GrammarThe 5th International Conference on Advanced Materials and ProcessingThe 6th International Conference on Signal Processing Proceedings 2002The 8th Biennial Conference on Tourism in AsiaTourism, Hospitality & Foodservice Industry2010 The 2nd IEEE International Conference on Information Management and EngineeringGroup 2International Conference on Infrared and Millimeter WavesInternational Conference on Molecular Beam EpitaxyInternational Conference on Narrow Gap SemiconductorsInternational Conference on Thin Film Physics and ApplicationInternational Conference on X-Ray LasersInternational Conference on Solid-state and Integrated Circuit Technology ProceedingsInternational Conference on High Performance Computer Architecture (HPCA) International Conference on Communications (ICC)International Joint Conference on Artificial Intelligence (IJCAI)International Conference on Computer Vision (ICCV)International Conference on Pattern Recognition (ICPR)International Conference on Pattern Recognition (ICPR)International Conference on Robotics and AutomationInternational Conference on Intelligent Robots and SystemsInternational Conference on Software EngineeringInternational Conference on Efficiency, Costs, Optimization, Simulation and Environmental Impact of Energy SystemsInternational Conference on Turbochargers and TurbochargingInternational Conference on Electrical MachinesInternational Technical Conference on the Enhanced Safety of Vehicles（EVS）International Conference on Magnet TechnologyInternational Conference on Engineering and Technological Sciences 2000 (PITTCON), USA.International Conference on Fluidization (ICF)International Conference on Bioseparation Engineering (ICBE)International Conference on Molecular Beam EpitaxyInternational Conference on ELT in ChinaInternational Conference on Pragmatics and Language LearningInternational Conference on MagnetismInternational conference on High Pressure Semiconductor PhysicsInternational Conference on Ternary and Multlnary CompoundsInternational Conference on Shellfish RestorationInternational Conference on Industrial Engineering and Engineering ManagementInternational Conference on Information SystemThe International Conference on MOVPEWorld Conference on WomenWorld Conference on Photovoltaic Solar Energy ConversionGroup 3IEEE International Conference on Accoustic, Speech and Signal Processing TechnologyIEEE International Conference on Accoustic, Speech and Signal Processing IFAC International Conference on Automatic ControlIEEE Conference on Decision and ControlThe IEEE International Conference on Vehicular Electronics and Safety (ICVES)The International IEEE Conference on Intelligent Transportation Systems (ITSC)IUMRS International Conference on Advanced MaterialsAnnual JALT International Conference on Language Teaching and Learning Asian Conference on Computer Vision (ACCV)Asian Conference on High Pressure ResearchThe Pacific Rim Conference on Lasers and Electro-Optics (CLEO/PR)Pan-Asian Conference on Language Teaching (PAC)Portland International Conference on Management of Engineering and TechnologyThe Pittsburgh Conference on Analytical Chemistry and Applied Spectroscopy Group 4The 4th World Conference for Graduate Research in Tourism, Hospitality and International Conference for Optics(光学)World Conference of Animal Production (WCAP)The 2nd International Conference of Bionic EngineeringⅡConference位于句首，有介词。

学科分类号0807本科毕业设计题目（中文）：水产自动投喂系统的设计（英文）：The design of feed system automaticIn aquatic products姓名: 张云涛学号: 2011180203院（系）: 工程与设计学院专业、年级 : 机械设计制造及自动化、2011级指导教师: 尹碧菊二〇一五年五月湖南师范大学本科毕业设计诚信声明本人郑重声明：所呈交的本科毕业设计，是本人在指导老师的指导下，独立进行研究工作所取得的成果，成果不存在知识产权争议，除设计中已经注明引用的内容外，本设计不含任何其他个人或集体已经发表或撰写过的作品成果。

对本设计的研究做出重要贡献的个人和集体均已在文中以明确方式标明。

本人完全意识到本声明的法律结果由本人承担。

本科毕业设计作者签名：二〇一五年五月六日湖南师范大学本科毕业设计任务书湖南师范大学工程与设计学院指导教师指导毕业设计情况登记表二、湖南师范大学本科毕业设计评审表优秀，80—89分记为良好，70—79分记为中等，60—69分记为及格，60分以下记为不及格。

若译文成绩为零，则不计总成绩，评定等级记为不及格。

三、湖南师范大学本科毕业设计答辩记录表目录水产自动投喂系统的设计机械设计制造及其自动化 2011级张云涛摘要随着我国水产养殖业的快速发展，大型标准化的池塘养殖也越来越多，因此在管理方面的工作量和复杂度也增加了许多。

目前，国内的大型水产养殖场投饵的自动化与智能化的普及率不高，仍然主要采用人工投饵的方式进行养殖。

但是与自动投饵相比，人工投饵存在着投饵量难以控制、人工劳动强度大、投喂饲料不均匀以及对生态坏境产生不良影响等缺点。

为了提高国内水产养殖投饵智能化、自动化和工业化的普及率，所以有必要设计并研发出一种符合大型标准化养殖投饵管理要求的自动投饵系统。

虽然目前的以单片机为控制核心的自动投饵机可以满足不同客户的各种需求，但也存在着工作时稳定度比较低、电动机发生故障的几率较高、无料时电机停止的可靠性不高和维修维护成本高等问题。

Abstr. Papers Am. Chem. Soc. Abstracts of Papers. American Chemical Society Acta Academiae Aboensis Acta Academiae AboensisActa Chem. Scand. Acta Chemica ScandinavicaActa Crystallograph. Acta CrystallographicaActa Hydrobiol. Sinica Acta Hydrobiolica SinicaActa Hydrochim. Hydrobiol. Acta Hydrochimica et HydrobiologicaActa Med. Scand. Acta Medica ScandinavicaActa Pharmaceutica Fennica Acta Pharmaceutica FennicaActa Scien. Circum. Acta Scientiae CircumstantiaeActa Vet. Brno Acta Veterinaria BrnoActa Vet. Scand. Acta Veterinaria ScandinavicaAdv. Cancer Res. Advances in Cancer ResearchAdv. Mar. Biol. Advances in Marine BiologyAgents Actions Agents and ActionsAgric. Gaz. N.S.W. Agricultural Gazette of New South WalesAlgae (Korean J. Phycol.) Algae (The Korean Journal of Phycology) Alzheimer's Rep. Alzheimer's ReportsAm. J. Botany American Journal of BotanyAm. J. Pathol. American Journal of PathologyAm. J. Pharmacy American Journal of PharmacyAm. J. Physiol.–Gastrointest. Liver Physiol. American Journal of Physiology-Gastrointestinal LiverPhysiologyAm. J. Pub. Health American Journal of Public HealthAm. J. Vet. Med. American Journal of Veterinary MedicineAm. J. Vet. Res. American Journal of Veterinary ResearchAm. Midland Nat. American Midland NaturalistAm. Rev. Resp. Dis. American Review of Respiratory DiseaseAmbio AmbioAnalyst AnalystAnalyt. Bioanalyt. Chem. Analytical and Bioanalytical ChemistryAnalyt. Biochem. Analytical BiochemistryAnalyt. Chem. Analytical ChemistryAnalyt. Chim. Acta Analytica Chimica ActaAnalyt. Lett. Analytical LettersAnalyt. Sci. Analytical SciencesAngew. Chem. Int. Ed. Angewandte Chemie–International EditionAnn. Allergy Annals of AllergyAnn. Internal Med. Annals of Internal MedicineAnn. Limnol. Annales de Limnologie–International Journal ofLimnologyAnn. N.Y. Acad. Sci. Annals. New York Academy of SciencesAnn. Rev. Microbiol. Annual Review of MicrobiologyAnn. Rev. Pharmacol. Toxicol. Annual Review of Pharmacology and Toxicology Appl. Biochem. Biotechnol. Applied Biochemistry and BiotechnologyAppl. Catalysis–B: Environ. Applied Catalysis. B. EnvironmentalAppl. Environ. Microbiol. Applied and Environmental MicrobiologyAppl. Microbiol. Applied MicrobiologyAppl. Microbiol. Biotechnol. Applied Microbiology and BiotechnologyAqua Fennica Aqua FennicaAquaculture AquacultureAquat. Ecol. Aquatic EcologyAquat. Ecosys. Health Manage. Aquatic Ecosystem Health & ManagementAquat. Microbial Ecol. Aquatic Microbial Ecology Proceedings of theConferenceAquat. Sci. Aquatic SciencesAquat. Toxicol. Aquatic ToxicologyArch. Anat. Micr. Morphol. Expér. Archives d’Anatomie Microscopique et de MorphologieExpérimentaleArch. Biochem. Biophys. Archives of Biochemistry and BiophysicsArch. Dermatol. Archives of DermatologyArch. Dermatol. Syphilol. Archives of Dermatology & SyphilologyArch. Exp. Veterinärmed. Archiv für Experimentelle VeterinärmedizenArch. Hydrobiol. Archiv für HydrobiologieArch. Hydrobiol.–Algol. Stud. Archiv für Hydrobiologie–Algological StudiesArch. Hydrobiol.–Beih. Ergebn. Limnol. Archiv für Hydrobiologie–Beiheft Ergebnisse derLimnologieArch. Hydrobiol.–Monogr. Beit. Archiv für Hydrobiologie–Monographische Beiträge Arch. Int. Pharmacodyn. Ther. Archives Internationales de Pharmacodynamie et deTherapieArch. Lat. Nutr. Archivos Latinoamericanos de NutricionArch. Microbiol. Archiv für MicrobiologieArch. Pharmacol. Archives of PharmacologyArch. Pharmazie Archiv der PharmazieArch. Toxicol. Archives of ToxicologyArizona Acad. Sci. Arizona Academy of Science. JournalArzneim.-Forsch.–Drug Res. Arzneimittel-Forschung–Drug ResearchAssoc. S.E. Biol. Bull. The Association of Southeastern Biologists Bulletin Aust. J. Biol. Sci. Australian Journal of Biological SciencesAust. J. Dairy Technol. Australian Journal of Dairy TechnologyAust. J. Ecol. Australian Journal of EcologyAust. J. Mar. Freshwater Res. Australian Journal of Marine and Freshwater Research Aust. NZ. J. Med. Australian and New Zealand Journal of Medicine Aust. NZ J. Pub. Health Australian and New Zealand Journal of Public Health Aust. Vet. J. Australian Veterinary JouranlAvian Dis. Avian DiseasesAvian Pathol. Avian PathologyBacteriol. Rev. Bacteriological ReviewsBiochem. BiochemistryBiochem. Biophys. Res. Commun. Biochemical and Biophysical Research Communications Biochem. J. Biochemical JournalBiochem. Pharmacol. Biochemical PharmacologyBiochem. Physiol. Pflanzen Biochemie und Physiologie der PflanzenBiochem. Soc. Trans. Biochemical Society. TransactionsBiochim. Biophys. Acta Biochimica et Biophysica ActaBiofizika BiofizikaBiofutur BiofuturBiogeochemistry BiogeochemistryBiol. Chem. Hoppe-Seyler Biological Chemistry Hoppe-SeylerBiol. Control Biological ControlBiol. Fertility Soils Biology and Fertility of SoilsBiol. Pharmaceut. Bull. Biological and Pharmaceutical Bulletin Biologia BiologiaBiological Res. 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British Journal of PharmacologyBr. Med. J. British Medical JournalBr. Phycol. J. British Phycological JournalBr. Poultry Sci. British Poultry ScienceBrain Res. Brain ResearchBrazilian J. Med. Biol. Res. Brazilian Journal of Medical and Biological Research Bull. Environ. Contam. Toxicol. Bulletin of Environmental Contamination and Toxicology Bull. Jap. Soc. Phycol. Bulletin. Japanese Society of PhycologyBull. Jap. Soc. Sci. Fish. Bulletin of the Japanese Society of Scientific Fisheries Bull. Korean Chem. Soc. Bulletin. Korean Chemical SocietyBull. Mar. Sci. Bulletin of Marine ScienceBull. Minn. Acad. Sci. The Bulletin of the Minnesota Academy of Science Bull. Natl. Sci. Mus. (Tokyo) Bulletin. National Science Museum (Tokyo)Bull. Pan Am. Health Org. Bulletin of the Pan American Health Organization Bull. Soc. Bot. France Bulletin de la Société Botaniqúe de FranceByull. Izobret. Byulleten IzobretiniiaCan. J. Anaes. Canadian Journal of AnaesthesiaCan. J. Biochem. Canadian Journal of BiochemistryCan. J. Biochem. Physiol. Canadian Journal of Biochemistry and Physiology Can. J. Bot. Canadian Journal of BotanyCan. J. Chem. Canadian Journal of ChemistryCan. J. Comp. Med. Canadian Journal of Comparative MedicineCan. J. Comp. Med. Vet. Sci. Canadian Journal of Comparative Medicine andVeterinary ScienceCan. J. Fish. Aquat. Sci. Canadian Journal of Fisheries and Aquatic Sciences Can. J. Microbiol. Canadian Journal of MicrobiologyCan. J. Pub. Health Canadian Journal of Public HealthCan. J. Zool. Canadian Journal of ZoologyCan. Med. Assoc. J. Canadian Medical Association. JournalCan. Pub. Health J. Canadian Public Heath JournalCan. Vet. J. Canadian Veterinary JournalCancer Chemother. Pharmacol. Cancer Chemotherapy and PharmacologyCancer Detect. Prevent. Cancer Detection and PreventionCancer Lett. Cancer LettersCancer Res. Cancer ResearchCancer Surv. Cancer SurveysCarcinogenesis CarcinogenesisCell. Biol. Toxicol. Cell Biology and ToxicologyCell Death & Diff. Cell Death & DifferentiationCell Growth & Diff. Cell Growth & DifferentiationCell. Mol. Biol. Lett. Cellular and Molecular Biology Letters Centr. Afr. J. Med. Central African Journal of Medicine Chem. Aust. Chemistry in AustraliaChem. Biol. Chemistry and Biology (London) Chem-Biol. Inter. Chemico-Biological Interactions Chem. Britain Chemistry in BritainChem. Commun. Chemical CommunicationsChem. Int. Chemistry InternationalChem. Pharmaceut. Bull. Chemical & Pharmaceutical Bulletin Chem. Res. Toxicol. Chemical Research in Toxicology Chem. Rev. Chemical ReviewsChem. Soc. Rev. Chemical Society Reviews Chemosphere ChemosphereChimia ChimiaChinese Biochem. J. Chinese Biochemical JournalChinese J. Analyt. Chem. Chinese Journal of Analytical Chemistry Chromatographia ChromatographiaCirc. Res. Circulation ResearchCirc. Shock Circulatory ShockClin. Allergy Clinical AllergyClin. Infect. Dis. Clinical Infectious DiseasesClin. Toxicol. Clinical ToxicologyCollect. Net The Collecting NetCommonw. Eng. Commonwealth EngineerComp. Biochem. Physiol.–B: Biochem. Mol.Biol. Comparative Biochemistry and Physiology. Part B.Biochemistry and Molecular BiologyComp. Biochem. Physiol.–C: Comp.Pharmacol. Comparative Biochemistry and Physiology. C.Comparative PharmacologyComp. Biochem. Physiol.–C: Pharmacol.Toxicol. Comparative Biochemistry and Physiology. C.Comparative Pharmacology and ToxicologyComp. Biochem. Physiol.–C: Pharmacol.Toxicol. Endocrinol. Comparative Biochemistry and Physiology. Part C.Pharmacology, Toxicology, and EndocrinologyContrib. N.S.W. Natl. Herb. Contributions. New South Wales National Herbarium Coral Reefs Coral ReefsCornell Vet. Cornell VeterinarianCountry Guide: Farm Mag. Country Guide: The Farm MagazineCrit. Rev. Plant Sci. Critical Reviews in Plant SciencesCrustaceana CrustaceanaCunninghamia CunninghamiaCurr. Biol. Current BiologyCurr. Med. Chem. Current Medicinal ChemistryCurr. Microbiol. Current MicrobiologyCurr. Opinion Microbiol. Current Opinion in MicrobiologyCurr. Opinion Struct. Biol. Current Opinion in Structural BiologyCurr. Pharmaceut. Design Current Pharmaceutical DesignCurr. Sci. Current ScienceCytobios CytobiosDansk Vet. Tidsskr. Dansk Veterinær TidsskriftDeep-Sea Res. Part 1 Deep-Sea Research Part 1–Oceanographic ResearchPapersDeut. Lebens.-Rund. Deutsche Lebensmittal-RundschauDev. Hydrobiol. Developments in HydrobiologyDis. Aquat. Org. Diseases of Aquatic OrganismsDNA Seq. DNA SequenceDokl. Akad. Nauk Doklady Akademii NaukDokl. Akad. Nauk SSSR Doklady Akademii Nauk SSSRDokl. Akad. Nauk. Ukr. SSR Ser. B Geol.Khim. Biol. Nauki Doklady Akademii Nauk Ukrainskoi SSR Ser B Geologicheskie Khimicheskie i Biologicheskie NaukiDokl. Biol. Sci. Doklady Biological SciencesDopov. Akad. Nauk. Ukr. RSR. Dopovidi Akademii Nauk Ukrainskoi RSRDrug Dev. Res. Drug Development ResearchDrug Metab. Rev. Drug Metabolism ReviewsEcol. Appl. Ecological ApplicationsEcol. Model. Ecological ModellingEcology EcologyEcotoxicol. Environ. Safety Ecotoxicology and Environmental Safety Electrochim. Acta Electrochimica ActaEMBO J. EMBO (European Molecular Biology Organization)JournalEndocrinology EndocrinologyEnviron. Carcinog. Ecotoxicol. Rev. Environmental Carcinogenesis and EcotoxicologyReviews. Part C. Journal of Environmental Science andHealthEnviron. Carcinog. Rev. Environmental Carcinogenesis Reviews. Part C. Journalof Environmental Science and HealthEnviron. Cons. Environmental ConservationEnviron. Health Perspect. Environmental Health PerspectivesEnviron. Int. Environment InternationalEnviron. Microbiol. Environmental MicrobiologyEnviron. Poll. Environmental PollutionEnviron. Res. Environmental ResearchEnviron. Rev. Environmental ReviewEnviron. Sci. Poll. Res. Environmental Science and Pollution Research Environ. Sci. Res. Environmental Science ResearchEnviron. Sci. Technol. Environmental Science and TechnologyEnviron. Technol. 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European Journal of Mass Spectrometry in Biochemistry,Medicine, and Environmental ResearchEvolution EvolutionExp. Bot. Experimental BotanyExp. Cell Res. Experimental Cell ResearchExperientia ExperientiaExpert Opin. Therapeut. Patents Expert Opinion Therapeutic PatentsFarming S. Afr. Farming in South AfricaFASEB J. FASEB JournalFauna FaunaFauna Flora Fauna och FloraFEBS Lett. FEBS (Federation of European Biochemical Societies)LettersFed. Proc. Federation ProceedingsFEMS Microbiol. Ecol. FEMS (Federation of European MicrobiologicalSocieties) Microbiology-EcologyFEMS Microbiol. Lett. FEMS (Federation of European MicrobiologicalSocieties) Microbiology LettersFEMS Microbiol. Rev. FEMS (Federation of European MicrobiologicalSocieties) Microbiology ReviewsField Stud. Field StudiesFinsk Tidskr. Finsk TidskriftFinsk Vet. Tidskr. Finsk VeterinärtidskriftFish Physiol. Biochem. Fish Physiology and BiochemistryFish. Sci. Fisheries ScienceFlicker FlickerFood Add. Contam. Food Additives and ContaminantsFood Agric. Immunol. Food and Agricultural ImmunologyFood Biotechnol. Food BiotechnologyFood Chem. Food ChemistryFood Chem. Toxicol. Food and Chemical ToxicologyForm. Kontrol Kach. Poverkhn. Vod Formirovanie i Kontrol Kachestva Poverkhnostnykh Vod Free Radical Res. Free Radical ResearchFresen. Environ. Bull. Fresenius Environmental BulletinFreshwater Biol. Freshwater BiologyFreskvandsfiskeribladet FreskvandsfiskeribladetFunctional Ecol. Functional EcologyGann (Jap. J. Cancer Res.) Gann (Japanese Journal of Cancer Research)Gazz. Chim. Ital. Gazzetta Chimica ItalianaGen. Pharmacol. General PharmacologyGene GeneGenetic Eng. Biotechnol. Genetic Engineer & Biotechnologist Gidrobiologicheskii Zh. (Hydrobiol. J.) Gidrobiologicheskii Zhurnal (Hydrobiological Journal) Gig. Sanit. Gigiana i SanitariiaHawaii Med. J. Hawaii Medical JournalHealth Environ. Digest Health & Environment DigestHelvetica Chim. Acta Helvetica Chimica ActaHepatology HepatologyHeterocycles HeterocyclesHuman Ecol. Risk Assess. Human and Ecological Risk AssessmentHuman Exp. Toxicol. Human & Experimental ToxicologyHuman Nature Human NatureHydrobiol. J. (Gidrobiologicheskii Zh.) Hydrobiological Journal (Gidrobiologicheskii Zhurnal) Hydrobiologia HydrobiologiaHydrol. Processes Hydrological ProcessesIEE Proc.–Sci. Measure. Technol. IEE Proceedings–Science, Measurement & Technology IMA. J. Math. Appl. Med. Biol. IMA (Institute of Mathematics and Its Applications)Journal of Mathematics Applied in Medicine &BiologyIndian J. Chem. Section A–Inorg. Bio-Inorg.Physical Theoret. Analyt. Chem. Indian Journal of Chemistry Section A–Inorganic Bio-Inorganic Physical Theoretical & Analytical ChemistryIndian J. Mar. Sci. Indian Journal of Marine SciencesInfect. Dis. Clin. Prac. Infectious Diseases in Clinical PracticeInt. Immunopharmacol. International ImmunopharmacologyInt. J. Environ. Analyt. Chem. International Journal of Environmental AnalyticalChemistryInt. J. Environ. Health Res. International Journal of Environmental Health Research Int. J. Epidemiology International Journal of EpidemiologyInt. J. Food Microbiol. International Journal of Food MicrobiologyInt. J. Immunopharmacol. International Journal of ImmunopharmacologyInt. J. Mass Spec. Ion Physics International Journal of Mass Specrometry & Ion Physics Int. J. Mol. Med. International Journal of Molecular MedicineInt. J. Peptide Protein Res. International Journal of Peptide and Protein Research Int. J. Pharmacog. International Journal of PharmacognosyInt. J. Sys Bacteriol. International Journal of Systematic BacteriologyInt. J. Sys. Evol. Microbiol. International Journal of Systematic & EvolutionaryMicrobiologyInt. Rev. Hydrobiol. International Review of HydrobiologyInt. Revue ges. Hydrobiol. Internationale Revue gesamten Hydrobiologie Internat. Verein. Limnol.Internationale Vereinigung für Theoretische undAngewandte LimnologieIowa State Coll. Vet. Iowa State College VeterinarianIsrael J. Plant Sci. Israel Journal of Plant SciencesIzv. Akad. Nauk. SSSR. Ser. Biol. Izvestiia Akademii Nauk SSSR, Seriia Biologicheskaia J. Adv. Oxidation Technol. Journal of Advanced Oxidation TechnologiesJ. Agric. Food Chem. Journal of Agricultural and Food ChemistryJ. Agric. S. Aust. Journal of Agriculture (South Australia)J. Agric. West. Aust. Journal of Agriculture of Western AustraliaJ. Allergy The Journal of AllergyJ. Allergy Clin. Immunol. Journal of Allergy and Clinical ImmunologyJ. Am. Chem. Soc. Journal. American Chemical SocietyJ. Am. Soc. Mass Spec. Journal of the American Society for Mass Spectrometry J. Am. Vet. Med. Assoc. Journal of the American Veterinary Medical Association J. Am. Water Works Assoc. Journal of the American Water Works AssociationJ. Analyt. Toxicol. Journal of Analytical ToxicologyJ. Antibiot. Journal of AntibioticsJ. Appl. Bacteriol. Journal of Applied BacteriologyJ. Appl. Bot. Journal of Applied Botany—Angewandte BotanikJ. Appl. Microbiol. Journal of Applied MicrobiologyJ. Appl. Microbiol. Symp. Suppl. Journal of Applied Microbiology Symposium Supplement J. Appl. Phycol. Journal of Applied PhycologyJ. Appl. Toxicol. Journal of Applied ToxicologyJ. Aquat. Animal Health Journal of Aquatic Animal HealthJ. Aquat. Plant Manage. Journal of Aquatic Plant ManagementJ. Assoc. Off. Analyt. Chem. Int Journal of AOAC (Association of Official AnalyticalChemists) InternationalJ. Aust. Inst. Agric. Sci. The Journal of the Australian Institute of AgriculturalScienceJ. Aust. Water Wastewater Assoc. Journal of the Australian Water and WastewaterAssociationJ. Bacteriol. Journal of BacteriologyJ. Basic Microbiol. Journal of Basic MicrobiologyJ. Biochem. Journal of BiochemistryJ. Biochem. Biophys. Methods Journal of Biochemical and Biophysical MethodsJ. Biochem. Toxicol. Journal of Biochemical ToxicologyJ. Biol. Chem. Journal of Biological ChemistryJ. Biosci. Bioeng. Journal of Bioscience and BioengineeringJ. Biotechnol. Journal of BiotechnologyJ. Cancer Res. Clin. Oncol. Journal of Cancer Research and Clinical Oncology J. Catalysis Journal of CatalysisJ. Cell Sci. Journal of Cell ScienceJ. Chart. Inst. Water Environ. Manage. Journal of the Chartered Institution of Water andEnvironmental ManagementJ. Chem. Ecol. Journal of Chemical EcologyJ. Chem. Soc., Chem. Commun. Journal. Chemical Society. Chemical Communications J. Chem. Soc. Chem. Comp. Journal. Chemical Society. Chemical CompoundsJ. Chem. Soc., Perkin Trans. 1 Journal of the Chemical Society–Perkin Transactions 1 J. Chromat. Journal of ChromatographyJ. Chromat. A Journal of Chromatography AJ. Chromat. B, Analyt. Technol. Biomed.Life Sci. Journal of Chromatography B, Analytical Technologies in the Biomedical and Life SciencesJ. Chromat. B: Biomed. Sci. Appl. Journal of Chromatography B: Biomedical Sciences andApplicationsJ. Clin. Microbiol. Journal of Clinical MicrobiologyJ. Colloid Interface Sci. Journal of Colloid and Interface ScienceJ. Colo. Wyo. Acad. Sci. Journal. Colorado-Wyoming Academy of ScienceJ. Computer-Aided Mol. Design Journal of Computer-Aided Molecular DesignJ. Computing Civil Eng. Journal of Computing in Civil EngineeringJ. Diarrhoeal Dis. Res. Journal of Diarrhoeal Diseases ResearchJ. Ecol. Journal of EcologyJ. Emerg. Med. Journal of Emergency MedicineJ. Environ. Biol. Journal of Environmental BiologyJ. Environ. Eng.–ASCE Journal of Environmental Engineering-ASCEJ. Environ. Manage. Journal of Environmental ManagementJ. Environ. Pathol. Toxicol. Oncol. Journal of Environmental Pathology, Toxicology &OncologyJ. Environ. Sci. Journal of Environmental SciencesJ. Environ. Sci. Health Part A–Environ. Sci.Eng.Journal of Environmental Science & Health Part A–Environmental Science & EngineeringJ. Environ. Sci. Health Part A–Toxic/ Hazardous Substances Environ. Eng. Journal of Environmental Science & Health Part A–Toxic/Hazardous Substances and Environmental EngineeringJ. Eukar. Microbiol. Journal of Eukaryotic MicrobiologyJ. Exp. Bot. Journal of Experimental BotanyJ. Exp. Mar. Biol. Ecol. Journal of Experimental Marine Biology and Ecology J. Fish Biol. Journal of Fish BiologyJ. Fish Dis. Journal of Fish DiseasesJ. Food Hygienic Soc. Japan Journal of the Food Hygienic Society of JapanJ. Freshwater Ecol. Journal of Freshwater EcologyJ. Gastroenterol. Hepatol. Journal of Gastroenterology and HepatologyJ. Gen. Microbiol. Journal of General MicrobiologyJ. Great Lakes Res. Journal of Great Lakes ResearchJ. Health Sci. Journal of Health ScienceJ. Health Toxicol. Journal of Health Toxicology (Chinese)J. High Resol. Chromat. Journal of High Resolution Chromatography andChromatography Communications (West Germany) J. Ind. Microbiol. Journal of Industrial MicrobiologyJ. Ind. Microbiol. Biotechnol. Journal of Industrial Microbiology & BiotechnologyJ. Infect. Dis. Journal of Infectious DiseasesJ. Infection Journal of InfectionJ. Inst. Water Eng. Sci. Journal of the Institution of Water Engineers & Scientists J. Inst. Water Environ. Manage. Journal of the Institution of Water and EnvironmentalManagementJ. Invert. Pathol. Journal of Invertebrate PathologyJ. Lake Sci. Journal of Lake SciencesJ. Limnol. Soc. S. Afr. Journal. Limnological Society of South AfricaJ. Liq. Chromat. Related Technol. Journal of Liquid Chromatography & RelatedTechnologiesJ. Mar. Biol. Assoc. U.K. Journal. Marine Biological Association (UnitedKingdom)J. Mass Spec. Journal of Mass SpectrometryJ. Mass Spec. Soc. Jap. Journal of the Mass Spectrometry Society of JapanJ. Med. Assoc. (Georgia) Journal of the Medical Association of GeorgiaJ. Med. Chem. Journal of Medicinal ChemistryJ. Med. Microbiol. Journal of Medical MicrobiologyJ. Med. Virol. Journal of Medical VirologyJ. Microbiol. Biotechnol. Journal of Microbiology and BiotechnologyJ. Microbiol. Methods Journal of Microbiological MethodsJ. Microcol. Separ. Journal of Microcolumn SeparationsJ. Microscopy Journal of Microcsopy–OxfordJ. Mol. Evol. Journal of Molecular EvolutionJ. Natl. Cancer Inst. Journal of the National Cancer InstituteJ. Natural Products Journal of Natural ProductsJ. Natural Toxins Journal of Natural ToxinsJ. Neurochem. Journal of NeurochemistryJ. Org. Chem. Journal of Organic ChemistryJ. Parasitol. Journal of ParasitologyJ. Pharmaceut. Biomed. Anal. Journal of Pharmaceutical and Biomedical AnalysisJ. Pharmaceut. Sci. Journal of Pharmaceutical SciencesJ. Pharmaceut. Soc. Jap. Journal of the Pharmaceutical Society of JapanJ. Pharmacol. Exp. Therapeut. Journal of Pharmacology and Experimental Therapeutics J. Photochem. Photobiol. A–Chem. Journal of Photochemistry and PhotobiologyA-ChemistryJ. Photochem. Photobiol. B–Biol. Journal of Photochemistry and Photobiology B-Biology J. Phycol. Journal of PhycologyJ. Planar Chromat. Journal of Planar ChromatographyJ. Plankton Res. Journal of Plankton ResearchJ. Plant Nutrition Journal of Plant NutritionJ. Porphyrins Phthalocyanines Journal of Porphyrins and PhthalocyaninesJ. S. Afr. Vet. Assoc. Journal of the South African Veterinary Association(Tydskrif van die Suid-Afrikaanse VeterinereVereniging)J. Sci. Ind. Res. Journal of Scientific and Industrial ResearchJ. Shellfish Res. Journal of Shellfish ResearchJ. Toxicol. Environ. Health Journal of Toxicology & Environmental HealthJ. Toxicol. Environ. Health–Part A Journal of Toxicology & Environmental Health–Part A J. Toxicol. Environ. Health–Part B Journal of Toxicology & Environmental Health–Part B J. Toxicol.–Toxin Rev. Journal of Toxicology-Toxin ReviewsJ. Trop. Med. Hygiene Journal of Tropical Medicine & HygieneJ. Vet. Diagn. Invest. Journal of Veterinary Diagnostic InvestigationJ. Vet. Med. B–Infect. Dis. Vet. Pub. Health Journal of Veterinary Medicine Series B-InfectiousDiseases and Veterinary Public HealthJ. Water Supply Res. Technol.-Aqua Journal of Water Supply Research & Technology–Aqua J. Wildlife Dis. Journal of Wildlife DiseasesJap. J. Cancer Res. Japanese Journal of Cancer ResearchJap. J. Cancer Res. (Gann) Japanese Journal of Cancer Research (Gann)Jap. J. Ecol. Japanese Journal of EcologyJap. J. Limnol. Japanese Journal of LimnologyJap. J. Toxicol. Environ. Health Japanese Journal of Toxicology and EnvironmentalHealthJersey J. The Jersey JournalKemia-Kemi Kemia-KemiLäkartidningen LäkartidningenLake Reservoir Manage. Lake and Reservoir ManagementLakeLine LakeLineLancet The LancetLC-GC Europe LC-GC EuropeLC-GC N. Am. LC-GC North AmericaLett. Appl. Microbiol. Letters in Applied MicrobiologyLett. Peptide Sci. Letters in Peptide ScienceLife Chem. Rep. Life Chemistry ReportsLife Sci. Pharmacol. Lett. Life Sciences Including Pharmacology Letters Limnol. (Berlin) Limnologica (Berlin)Limnol. Oceanogr. Limnology and OceanographyLimnologica LimnologicaLimnology LimnologyLipids LipidsLouisiana Municipal Rev. Louisiana Municipal Review (Shreveport)Magnetic Resonance Chem. Magnetic Resonance in Chemistry and Biology. Based onLectures at the Ampere International Summer School Magy. Állator. Lap. Magyar Állatorvosok LapjaMar. Biol. Marine BiologyMar. Ecol.–Prog. Ser. Marine Ecology. Progress SeriesMar. Freshwater Res. Marine & Freshwater ResearchMar. Poll. Bull. Marine Pollution BulletinMaryland Med. J. Maryland Medical JournalMass Spec. Rev. Mass Spectrometry ReviewsMed. J. Aust. Medical Journal of AustraliaMed. Microbiol. Immunol. Medical Microbiology and ImmunologyMed. Vet. Entomol. Medical Veterinary EntomologyMedicina Clinica Medicina ClinicaMem. Inst. Osw. Cruz Memorias do Instituto Oswaldo CruzMh. Vet. Med. Monatshefte fuer VeterinaermedizinMicrobial Ecol. Microbial EcologyMicrobiol. Immunol. Microbiology and ImmunologyMicrobiol. Rev. Microbiological ReviewsMicrobiol. Sci. Microbiological SciencesMicrobiology Microbiology (Mikrobiologiya)Microbiology–SGM Microbiology–Society for General Microbiology Microbios MicrobiosMicrobios Lett. Microbios LettersMicroscopy Res. Technique Microscopy Research and TechniqueMikrobiol. Zh. Mikrobiologicheskii Zh.Mikrokosmos MikrokosmosMilk Producer The Milk ProducerMinn. Botan. Ser. IV Minnesota Botanical Series IVMitt. FischVer. Prov. Brandenburg Mitteilungen der Fischereivereine für die ProvinzBrandenburg。

智能农业论文参考文献一、智能农业论文期刊参考文献[1].基于物联网的英国智能农业进展研究.《安徽农业科学》.被中信所《中国科技期刊引证报告》收录ISTIC.被北京大学《中文核心期刊要目总览》收录PKU.2014年19期.彭英.陈楠.施小飞.[4].物联网技术与江苏智能农业产业发展.《江苏农业科学》.被中信所《中国科技期刊引证报告》收录ISTIC.被北京大学《中文核心期刊要目总览》收录PKU.2011年5期.戴起伟.凡燕.曹静.朱科峰.王支凤.[5].基于物联网的智能农业管理系统研究与设计——以江苏省农业物联网平台为例.《江苏农业科学》.被中信所《中国科技期刊引证报告》收录ISTIC.被北京大学《中文核心期刊要目总览》收录PKU.2013年5期.刘家玉.周林杰.荀广连.吴爱民.陈磊.[6].基于ECDSA优化算法的智能农业无线传感器节点的网络安全认证. 《江苏农业科学》.被中信所《中国科技期刊引证报告》收录ISTIC.被北京大学《中文核心期刊要目总览》收录PKU.2015年4期.马少华.张兴.韩冬.史伟.[7].基于GrayEKF算法的智能农业车辆同时定位与地图创建.《农业工程学报》.被中信所《中国科技期刊引证报告》收录ISTIC.被EI收录EI.被北京大学《中文核心期刊要目总览》收录PKU.2012年19期.田光兆.安秋.姬长英.顾宝兴.王海青.赵建东.[8].基于ZigBee的智能农业管理系统设计.《湖北农业科学》.被中信所《中国科技期刊引证报告》收录ISTIC.被北京大学《中文核心期刊要目总览》收录PKU.2013年3期.滕志军.何建强.李国强.[9]基于立体视觉的智能农业车辆实时运动检测.《农业机械学报》.被中信所《中国科技期刊引证报告》收录ISTIC.被EI收录EI.被北京大学《中文核心期刊要目总览》收录PKU.2013年7期.田光兆.安秋.姬长英.顾宝兴.王海青.赵建东.[10].基于物联网技术的智能农业种植系统设计.《现代电子技术》.被中信所《中国科技期刊引证报告》收录ISTIC.2013年24期.宋艳.二、智能农业论文参考文献学位论文类[1].智能农业平台开发.被引次数：1作者：瞿韬.信号与信息处理浙江理工大学2014（学位年度）[2].基于全信息的智能农业装备技术专利战略研究.被引次数：1作者：赵旭.情报学江苏大学2010（学位年度）[3].基于物联网的智能农业大棚的研究与实现.作者：初洪龙.计算机应用技术大连理工大学2014（学位年度）[4].物联网在智能农业中的应用研究.被引次数：11作者：彭改丽.计算机技术郑州大学2012（学位年度）[5].基于物联网的智能农业监测系统的设计与实现.被引次数：11作者：王冬.通信与信息系统大连理工大学2013（学位年度）[6].基于Web的智能农业远程监控系统的研究.作者：张华飞.计算机技术浙江理工大学2015（学位年度）[7].嵌入式农作物生境监测与智能决策系统研究.作者：尤聚军.检测技术与自动化装置中国科学技术大学2013（学位年度）[8].远程数据采集在智能农业中的应用——基于K60和GPRS的设计.被引次数：1作者：蒋婷.计算机应用技术苏州大学2014（学位年度）[9].基于嵌入式Web技术的温室环境在线监测与控制系统.作者：周超.测试计量技术及仪器厦门大学2014（学位年度）[10].智能农业系统中感知适配网关的研究与设计.被引次数：5作者：李萍.信号与信息处理浙江理工大学2012（学位年度）三、相关智能农业论文外文参考文献[1]Implementationofintelligentairconditionerforfineagriculture. MingTaYangChengChuanChenYenLongKuo《Energyandbuildings》,被EI收录EI.被SCI收录SCI.2013May[2]Anautonomousintelligentgatewayinfrastructureforinfieldprocessi nginprecisionviticulture..Peres,E.Fernandes,M.A.Morais,R.Cunha,C.R.Lopez,J.A.Matos,S.R.Ferreira ,P.J.S.G.Reis,M.J.C.S.《ComputersandElectronicsinAgriculture》,被EI收录EI.被SCI收录SCI.20112[3]AUDIOVISUALRECOGNITIONOFGOOSEFLOCKINGBEHAVIOR. KIMARILDSTEENOLEROLANDTHERKILDSENOLEGREENHENRIKKARSTOFT 《Internationaljournalofpatternrecognitionandartificialintelligence》,被EI收录EI.被SCI收录SCI.20137[4]NeuralIntelligentWaterDropsalgorithmtoselectrelevanttexturalfe aturesfordevelopingprecisionirrigationsystemusingmachinevision.. Hendrawan,Y.Murase,H.《ComputersandElectronicsinAgriculture》,被EI收录EI.被SCI收录SCI.20112[5]AnIntelligentAmmoniaSensorBasedonMultiParameterforAquaculture. HaijiangTaiQishengDingLihuaZengShuangyinLiuDaoliangLi 《SensorLetters:AJournalDedicatedtoallAspectsofSensorsinScience,Engin eering,andMedicine》,被EI收录EI.被SCI收录SCI.20136/7[6]DeterminationofthiophanatemethylusingUVabsorptionspectrabasedo nmultiplelinearregression.L.JiaoD.DongW.ZhengX.ZhaoS.ZhangC.Shen《Optik:ZeitschriftfurLichtundElektronenoptik:=JournalforLightandElec tronoptic》,被EI收录EI.被SCI收录SCI.20141[7]INTELLIGENTINFORMATIONTECHNOLOGYINAGRICULTURE. DongRenChunjiangZhaoSimonX.Yang 《Intelligentautomationandsoftcomputing》,被SCI收录SCI.20133[8]Designofintelligentgreenhouseenvironmentmonitoringsystembasedo nZigBeeandembeddedtechnology.WeiminQiuLinxiDongFeiWangHaixiaYan2014[9]AnPortableIntelligentMeasurementInstrumentforDissolvedOxygenin Aquaculture.LinlinZhaoDaoliangLiDaokunMaQishengDing《SensorLetters:AJournalDedicatedtoallAspectsofSensorsinScience,Engin eering,andMedicine》,被EI收录EI.被SCI收录SCI.20101[10]ANeuroDynamicProgrammingBasedOptimalControllerforTomatoSeedli ngGrowthinGreenhouseSystems.J.PUCHETAH.PATINOR.FULLANAC.SCHUGURENSKYB.KUCHEN 《Neuralprocessingletters》,被EI收录EI.被SCI收录SCI.20063四、智能农业论文专著参考文献[1]智能农业复杂大系统控制及建模.陈一飞.郑伟.杜尚丰.张静，20112011年中国智能自动化会议[2]物联网技术与江苏智能农业产业发展.戴起伟.曹静.凡燕.朱科峰.王支凤，2011中国畜牧兽医学会信息技术分会2011年学术研讨会[3]基于智能农业理念的草莓立体栽培生产管控大系统构建.陈一飞.陈卫文.齐凯.杜尚丰.董乔雪，2011中国农业工程学会2011年学术年会[4]智能农业管理信息系统[6]基于NC的智能农业应用系统研究.叶丽.吴华瑞.孙想，20032003中国计算机大会[7]智能农业及其综合环境的开发.杨一平，20032003年中国数字农业与农村信息化发展战略研讨会[8]农业信息化技术研究进展综述.郑国清，2002中国农学会青年农业科学学术年会[9]广东数字农业实施模式的探讨.赵新.罗锡文.任春涛.2005年中国农业工程学会学术年会[10]日光温室集群大系统的复杂性与系统控制.陈一飞.齐凯.刘柏成.杜尚丰，20132013年中国智能自动化会议。

Ž.Aquaculture1801999147–165www.elsevier.nl r locate r aqua-onlineReviewThe use of probiotics in aquacultureF.J.Gatesoupe)´Unite Mixte de Nutrition des Poissons INRA-IFREMER,IFREMER,Centre de Brest,BP70,F-29280´Plouzane,FranceAccepted12April1999AbstractThe research of probiotics for aquatic animals is increasing with the demand for environment-friendly aquaculture.The probiotics were defined as live microbial feed supplements that improve health of man and terrestrial livestock.The gastrointestinal microbiota of fish and shellfish are peculiarly dependent on the external environment,due to the water flow passing through the digestive tract.Most bacterial cells are transient in the gut,with continuous intrusion of microbes coming from water and food.Some commercial products are referred to as probiotics,though they were designed to treat the rearing medium,not to supplement the diet.This extension of the probiotic concept is pertinent when the administered microbes survive in the gastrointestinal tract. Otherwise,more general terms are suggested,like biocontrol when the treatment is antagonistic to pathogens,or bioremediation when water quality is improved.However,the first probiotics tested in fish were commercial preparations devised for land animals.Though some effects were observed with such preparations,the survival of these bacteria was uncertain in aquatic environ-ment.Most attempts to propose probiotics have been undertaken by isolating and selecting strains from aquatic environment.These microbes were Vibrionaceae,pseudomonads,lactic acid bacteria, Bacillus spp.and yeasts.Three main characteristics have been searched in microbes as candidatesŽ.to improve the health of their host.1The antagonism to pathogens was shown in vitro in most Ž.Ž.cases.2The colonization potential of some candidate probionts was also studied.3Challenge tests confirmed that some strains could increase the resistance to disease of their host.Many other beneficial effects may be expected from probiotics,e.g.,competition with pathogens for nutrients or for adhesion sites,and stimulation of the immune system.The most promising prospects are sketched out,but considerable efforts of research will be necessary to develop the applications to aquaculture.q1999Elsevier Science B.V.All rights reserved.Keywords:Microbial ecology;Digestive tract;Feed additives;Disease control;Antibacterial activity;Water quality)Tel.:q33-2-9822-4389;fax:q33-2-9822-4653;E-mail:joel.gatesoupe@ifremer.fr0044-8486r99r$-see front matter q1999Elsevier Science B.V.All rights reserved.Ž.PII:S0044-84869900187-8()148F.J.Gatesoupe r Aquaculture1801999147–1651.IntroductionLong before their discovery,microbes have been unawarely used to preserve food,Ž. and these empirical methods contributed to improve human health Bengmark,1998.Ž.Ž.Early in the century,Metchnikoff1907,1908cited by Tannock,1997proposed to implant lactic acid bacteria into the human intestine,with a view to suppressing the detrimental activity of other microbes.The modern concept of probiotics was formulated Ž.only25years ago Parker,1974,then its pertinence was challenged for many years among the scientific community.Several definitions of probiotics were successively Ž.proposed.Parker1974originally referred to‘‘organisms and substances which con-tribute to intestinal microbial balance’’as probiotics.The definition was then restricted to‘‘a live microbial feed supplement which beneficially affects the host animal byŽ.Ž. improving its intestinal microbial balance’’Fuller,1989.Tannock1997noted that the effect on the‘‘intestinal microbial balance’’has not been demonstrated in most cases,and he proposed to speak of‘‘living microbial cells administered as dietary supplements with the aim of improving health’’.To my knowledge,the first application of probiotics in aquaculture seems relatively Ž.recent Kozasa,1986,but the interest in such environment-friendly treatments is increasing rapidly.Scientific evaluation corroborated seldom the first empirical trials, and the information was mainly spread by‘‘grey literature’’.However,a growing number of scientific papers have dealt explicitly with probiotics,and it is now possible to survey the state of the art,from the empirical use to the scientific approach.AŽprevious review was devoted to lactic acid bacteria in finfish Ringøand Gatesoupe, .1998,but many other microbes have been tested as probiotics for various aquatic animals,appealing for a general overview.It is essential to remind some definitions of ecological concepts to clear away theŽ. treatments termed improperly‘‘probiotic’’.The goals of this paper are1to examineŽ.the pertinence of such terminology applied to the aquatic environment,2to draw theŽ.different trends of applications,and3to point out needs for further research.2.Is the intestinal environment of aquatic animals favourable to probiotics?Aquatic animals are quite different from the land animals for which the probiotic concept was developed,and a preliminary question is the pertinence of probiotic applications to aquaculture.Man and terrestrial livestock undergo embryonic development within an amnion, whereas the larval forms of most fish and shellfish are released in the external environment at an early ontogenetic stage.These larvae are highly exposed to gastro-intestinal microbiota-associated disorders,because they start feeding even though theŽ.digestive tract is not yet fully developed Timmermans,1987,and though the immuneŽ.system is still incomplete Vadstein,1997.Thus,probiotic treatments are particularly desirable during the larval stages.()F.J.Gatesoupe r Aquaculture1801999147–165149Gram-positive obligate or facultative anaerobes are dominant in the gastrointestinalŽ. microbiota of man and terrestrial farm animals Gournier-Chateau et al.,1994.Inhuman feces,the major bacterial groups are Bacteroides,Gram-positive anaerobic cocci,Ž.Eubacterium,and Bifidobacterium Hume,1997,whereas the predominant groups inŽ.pig feces are‘‘streptococci’’and‘‘lactobacilli’’Stewart,1997.Most probionts belongto dominant or sub-dominant genera among these microbiota,e.g.,Bifidobacterium,Ž.Lactobacillus,Streptococcus Gournier-Chateau et al.,1994.Gram-negative facultativeanaerobes prevail in the digestive tract of fish and shellfish,though symbiotic anaerobesŽmay be dominant in the posterior intestine of some herbivorous tropical fish Clements, .Ž1997.Vibrio and Pseudomonas are the most common genera in crustaceans Moriarty, .Ž.Ž.1990,marine fish Sakata,1990and bivalves Prieur et al.,1990.Aeromonas,Ž. Plesiomonas and Enterobacteriaceae are dominant in freshwater fish Sakata,1990.Aconsequence of the specificity of aquatic microbiota is that the most efficient probioticsfor aquaculture may be different from those of terrestrial species.The resident microbes benefit from a fairly constant habitat in the gastrointestinaltract of man and terrestrial livestock,whereas most microbes are transient in aquatic Ž.animals Moriarty,1990.These animals are poikilothermic,and their associated micro-Ž.´biota may vary with temperature changes Lesel,1990.Salinity changes may also Ž. influence microbiota Hamid et al.,1978;Sakata et al.,1980;Ringøand Strøm,1994,and marine finfish are obliged to drink constantly to prevent water loss from the body.This continuous water flow increases the influence of the surrounding medium,in thesame way as the water flow observed in filter-feeders,like bivalves,shrimp larvae andlive food organisms.This influence is particularly important in larvae,when the gastricbarrier is absent.Therefore,the intestinal microbiota of aquatic animals may changerapidly with the intrusion of microbes coming from water and food.In bivalves,theŽassociated microbiota is very similar to those found in seawater and sediment Sugita et.al.,1981;Prieur et al.,1990.The same kinds of bacteria were found in the gut ofPenaeus japonicus and in seawater,but normal members of microbiota may beŽ.introduced via the diet Moriarty,1990.In larval and juvenile fish,the influence of foodŽ. has been clearly demonstrated Tanasomwang and Muroga,1989;Ringøet al.,1995. The influence of bacteria brought by live food organisms is particularly dramatic during Ž.first feeding Munro et al.,1993;Bergh et al.,1994.3.Applications of the‘‘probiotic’’concept in a broad senseThe transience of aquatic microbes may legitimate the extension of the probioticŽ. concept to living microbial preparations used to treat aquaculture ponds.Moriarty1998 proposed to extend the definition of probiotics to microbial‘‘water additives’’.How-Ž.ever,this extension would make too vague definition of Tannock1997.I suggest analternative definition of probiotics as:microbial cells that are administered in such a wayas to enter the gastrointestinal tract and to be kept alive,with the aim of improvinghealth.This latter definition will be used in the following to sort the microbialpreparations which can be designated as probiotics.()150F.J.Gatesoupe r Aquaculture1801999147–165In1991,Porubcan reported on two attempts at bacterial treatments to improve waterŽ.quality and production yield of Penaeus monodon.1Floating biofilters pre-inoculated with nitrifying bacteria decreased the amounts of ammonia and nitrite in the rearingŽ.Ž.water.This treatment increased shrimp survival Porubcan,1991a.2The introduction of Bacillus spp.in proximity to pond aerators reduced chemical oxygen demand,andŽ.increased shrimp harvest Porubcan,1991b.Several commercial products have sought to exploit the same idea that bacteria which improve water quality may be beneficial to Žanimal health e.g.,‘‘bacteria’’,Aquatic Warehouse,San Diego,CA;‘‘Biostart’’, Advanced Microbial Systems,Shakopee,MN;‘‘BRF-1A,BRF-13A,PB-32,PBL-44’’, Enviro-Reps International,Camarillo,CA;‘‘Liqualife’’,Cargill,Animal Nutrition Divi-sion;‘‘microbial and enzymic products’’,Alliance Bioremediation and Composting, Encinitas,CA;‘‘PondPro-VC’’,Biomanagement Systems,Wellington Point,Australia;.‘‘probiotics’’,Contessa,ZB Industries,San Pedro,CA.These products are referred to as‘‘probiotics’’and most of them contain nitrifying bacteria and r or Bacillus spp. These two kinds of bacteria are quite different.The nitrifying bacteria have strict ecological niches,and they have not been detected in the gastrointestinal tract of animals.The strains of Bacillus ed as probiotics for terrestrial livestock have telluric origins,and they are not autochthonous in the gastrointestinal tract,but they mayŽ.be active during intestinal transit Gournier-Chateau et al.,1994.Moreover,there areŽmany reports of isolation of Bacillus strains from fish Hamid et al.,1978;Strøm and Olafsen,1990;Nedoluha and Westhoff,1995;Sadhukhan et al.,1997;Kennedy et al.,.Ž1998;Sugita et al.,1998,crustaceans Austin and Allen,1982;Sharmila et al.,1996;.Ž.Ž. Sugita et al.,1996a,and bivalves Sugita et al.,1981.Queiroz and Boyd1998 confirmed that a commercial inoculum of Bacillus spp.increased survival and produc-tion of channel catfish,but these authors focused their attention on water quality criteria,Ž.which were poorly affected by the treatment.Kennedy et al.1998isolated a strain of Bacillus subtilis from the common snook,Centropomus undecimalis.The inoculation of this strain into the rearing water resulted in the apparent elimination of Vibrio sp.from whole larvae of snook,after decreasing salinity from ca.30to ca.3practical salinity Ž.units.Moriarty1998noted an increase of prawn survival in ponds where some strains of Bacillus spp.were introduced.This treatment decreased the proportion of pathogenic luminous Vibrio spp.in the sediments,and to a lesser extent,in the water.The effect on prawn intestinal microbiota was not studied.The strains of Bacillus were selected because of their antibiotic activity against luminescent Vibrio sp.,but the author emphasized the multiplicity of the possible probiotic effects,e.g.,enzymatic excretions, competition for nutrients and for space.These various mechanisms of action might prevent the emergence of resistant strains,a well-known risk of antibiotic treatments. However,these speculations need to be confirmed by experimental evidence,in particu-lar,the reference to the principle of competitive exclusion,frequently put forward in advertisements for probiotics.This principle is based on the observation that experi-ments in chemostats‘‘usually result in the replacement of all members of an initial community by one organism best adapted to those highly selective substrate-limited Ž.growth conditions’’Dolfing and Gottschal,1997.The antibiotic activity of the strains of Bacillus would interfere,making uneasy to demonstrate competitive exclusion in thisŽ.case.The actual data of Moriarty1998showed the inhibitory activity of Bacillus spp.()F.J.Gatesoupe r Aquaculture1801999147–165151 against luminous Vibrio sp.in pond sediment,but the effect on prawn survival might be due either to a probiotic effect,or to an indirect effect on animal health.For instance,the degradation of organic matter by Bacillus spp.might improve water quality.Therefore, the use of Bacillus spp.as pond supplement needs further investigation to be considered as a probiotic treatment,with particular attention to a possible intestinal transience.The probiotic treatments may be considered as methods of biological control,the so-called‘‘biocontrol’’that termed the limitation or the elimination of pests by the introduction of adverse organisms,like parasites or specific pathogens.Maeda et al.Ž.1997proposed to designate as biocontrol the methods of treatment using‘‘theŽ.antagonism among microbes...through which pathogens can be killed or reduced in number in the aquaculture environment’’.In this acceptation,the pond treatmentŽ.proposed by Moriarty1998is indisputably relevant to biocontrol,as well as many other microbial treatments,including those whose target organisms are not animals,but Ž.microalgae Rico-Mora et al.,1998.Another terminology should designate the applica-tions of nitrifying bacteria that are related to the bioremediation concept.This concept refers to the treatment of pollutants or waste by the use of microorganisms that breakŽ.down the undesirable substances Fig.1.The same concept is sometimes named Ž.bioaugmentation Moriarty,1997,1998.Fig.1.Tentative classification of microbial treatments used in aquaculture,according to current terminology.Ž. The term‘‘probiotics’’is reserved to strains transient or resident in the gastrointestinal tract Section3,Ž.‘‘biocontrol’’implies only that the strains are antagonistic to pathogens Maeda et al.,1997,and‘‘bioremedi-Ž.ation’’refers to breakdown of pollutants or waste by the microbes Moriarty,1997,1998.()152F.J.Gatesoupe r Aquaculture1801999147–1654.Application of commercial products for terrestrial livestockThe first trials of incorporation of probiotics into aquaculture feeds used commercialpreparations designed for land animals.Spores of Bacillus toyoi isolated from soilŽreduced the mortality of Japanese eel which were infected by Edwardsiella sp.,Kozasa, .Ž. 1986.The same feed additive increased the growth rate of yellowtail Kozasa,1986.Spores may be easily incorporated into compound food,but their fate in the gastrointes-tinal tract of fish was not followed in these experiments.It would be particularlyinteresting to know whether the spores may germinate in the gut,depending on transitŽ. time and rearing temperature.The same strain of B.toyoi used by Kozasa1986was later tested on rotifers,Brachionus plicatilis,which were left to filter the spores for2h Ž.Gatesoupe,1989.This treatment increased the growth rate of larval turbot,but themicrobiota were studied neither in the larvae nor in the ter,a study wasperformed with the food additive‘‘Paciflor9’’containing spores of Bacillus IP5832bycounting and characterising bacteria associated with spore-fed rotifers and turbot larvae Ž.Gatesoupe,1993.Most spores of Bacillus sp.were filtered by the rotifers within lessthan half an hour,but the number of cultivable cells of the strain decreased sharply inthe rotifers1h after introduction of the spores into the water.These cells were thusrecovered alive in the rotifers,but for a period probably too short to allow an actualŽ.probiotic effect,according to the definition suggested above Section3.Many bacilliŽ. produce antibiotics,especially in relation to the sporulation process Brock,1974,andŽsome antibiotics may be produced by proteolysis of the vegetative cells Vitkovic and .Sadoff,1977.When rotifers were fed with spores,the decrease of the Vibrionaceaenormally dominant in the rotifers might be due to such a release of antibiotic from the Ž.cells of Bacillus sp.Gatesoupe,1993.Few cultivable cells of the strain of Bacillus sp.were recovered from turbot fed for five days with the spore-treated rotifers.A directprobiotic effect was therefore unlikely,though this treatment increased the resistance ofturbot larvae exposed to pathogenic Vibrio sp.Commercial preparations with live lactic acid bacteria have also been introduced intothe medium of live food organisms for larval flatfish.Some of these treatmentsincreased the production of rotifers and the growth of turbot and Japanese flounder Ž.Gatesoupe,1989,1991;Gatesoupe et al.,1989.Some preparations with lactic acidbacteria limited also the proliferation of bacteria in rotifers,but the fate of the lactic acidŽ. bacteria was not studied in these experiments Gatesoupe et al.,1989;Gatesoupe,1991. Other commercial preparations of Streptococcus faecium improved the growth and feedŽ.efficiency of Israeli carp Noh et al.,1994;Bogut et al.,1998.Escherichia colidisappeared from the intestinal microbiota of carp after14days of feeding with theŽ.probiotic preparation Bogut et al.,1998.These authors stated that S.faecium‘‘hashigh adhesive ability in the epithelium of carp digestive tract’’,but without anyexperimental evidence.These trials with commercial probiotics for land animals were important to show theinterest of bacterial additives in aquaculture feeds,but the survival of these microbeswas uncertain in the gastrointestinal tract of aquatic animals.Most attempts have beenafterwards aimed at seeking autochthonous strains with probiotic properties.()F.J.Gatesoupe r Aquaculture1801999147–1651535.The search for autochthonous aquatic probiotics5.1.Isolation and characterization of autochthonous microbesIn juvenile fish and shellfish,the autochthonous microbes may be isolated from the digestive tract after dissection,while distinguishing stomach and intestine regions.The microbes adherent to epithelial cells can be separated from those adherent to mucus,andŽ.from those transient in the lumen Westerdahl et al.,1991.These methods are not applicable to larvae and live food organisms,but the external surface of larval fish may be washed with a0.1%benzalkonium chloride saline solution to differentiate theŽmicrobes adherent to the external surface from those present in the gut Blanch et al., .Ž1997.Many microbes may be isolated on selective media Pratt and Reynolds,1973;.Flint,1985;Jeppesen,1995;Donovan and van Netten,1995.Then the isolates areŽcharacterized by proper methods Roth et al.,1962;Hansen and Sorheim,1991;Holt et.al.,1994;Bertone et al.,1996;Austin et al.,1997;Tannock,1999.5.2.Pioneering studiesŽ.The first successful report seems to be attributed to Maeda and Liao1992,who isolated a strain‘‘PM-4’’from the rearing water of larval Pen.monodon,with goodŽsurvival and molting rate.The bacterium,identified as Thalassobacter utilis Nogami et .Žal.,1997,was used for the biocontrol of the larval rearing of Pen.monodon Maeda.and Liao,1992;Maeda et al.,1997and the swimming crab,Portunus trituberculatus Ž.Nogami and Maeda,1992;Nogami et al.,1997.This biocontrol treatment increasedŽthe survival of the larvae,and repressed the growth of Vibrio anguillarum Nogami and .Ž. Maeda,1992and Haliphthoros sp.fungus,Lagenidales;Nogami et al.,1997.It would be worth studying whether T.utilis can survive in the gut of the larval crab,sinceŽV.anguillarum infection can start via the intestinal route Colorni,1985;Grisez et al.,.1996;Olsson et al.,1996;Garcia et al.,1997.Ž.Griffith1995reported that shrimp larvae reared in Ecuadorian hatcheries were affected by a disease characterized by a change in the bacterial population.The proportion of Vibrio alginolyticus decreased,whereas Vibrio parahaemolyticus in-creased.The strain of V.alginolyticus was isolated and used as probiotic in many hatcheries,where shrimp survival was restored to the level obtained before diseaseŽ.outbreak.Austin et al.1995investigated the probiotic effect of this strain,and these authors reported that cells of Vibrio ordalii lost their viability within3h after the introduction of freeze-dried supernatant of probiotic culture into the suspension medium. V.anguillarum and Aeromonas salmonicida were also inhibited,but to a lesser extent. The probiont survived in the intestine of Atlantic salmon for at least3weeks,and a preliminary bath with this probiont improved the survival of salmon challenged withŽ. pathogens.This provides an example of what might be expected from probiotics:1Ž.antagonism to pathogens,2gut colonization,with possible adhesion to intestinal Ž.mucus,and3increased resistance of the host to pathogens.()154F.J.Gatesoupe r Aquaculture1801999147–1655.3.Antagonism to pathogensŽ.Antagonism seems to be common among marine bacteria Table1.For example,Ž.over60%of isolates from zooplankton were bacteriolytic Nair et al.,1985,and up toŽ75%of the isolates from sponges produced antibacterial compounds Marty and Martin, .Ž. 1992.A pathogenic Vibrio sp.was inhibited by a variable proportion0–100%of theŽ.bacteria isolated from first-feeding halibut larvae Bergh,1995.Most marine ant-Table1Antagonism of aquatic microbes to fish and shellfish pathogens.Ae.:Aeromonas;Ae.h.:Ae.hydrophila; Ae.s.:Ae.salmonicida;Ed.t.:Edwardsiella tarda;En.s.:Enterococcus seriolicida;IHNV:infectious hemato-poietic necrosis virus;OMV:Onchorhynchus masou virus;Ps.d.:Ps.doudorofii;Pa.p.:Pasteurella piscicida;V.:Vibrio;V.al.:V.alginolyticus;V.an.:V.anguillarum;V.o.:V.ordalii;Y.r.:Yersinia ruckeri Antagonist Source Pathogens tested ReferenceŽ. Freshwater bacteria fish intestine Aeromonas spp.Sugita et al.,1996bŽ. Freshwater bacteria salmonid hatchery IHNV Kamei et al.,1988Ž. Marine bacteria invertebrates Vibrio spp.Marty and Martin,1992Ž. Marine bacteria seaweeds Ed.t.,Pa.p.,Ae.spp.,Dopazo et al.,1988V.spp.,Y.r.Ž. Marine bacteria Scophthalmus maximus Ae.h.,Ae.salmonicida,V.an.Westerdahl et al.,1991Ž. Marine bacteria various Ae.h.,V.an.Ivanova et al.,1998Ž. Marine bacteria various En.s.;Pa.p.;V.an.;Sugita et al.,1996aVibrioÕulnificusŽ. Marine bacteria various IHNV Kamei et al.,1987Ž. Marine bacteria various‘‘V.anguillarum-related’’Riquelme et al.,1997Ž. Aeromonas media seawater Ae.spp.,V.spp.,Y.r.Gibson et al.,1998Ž. Alteromonas Argopecten purpuratus Ae.h.,V.al.,V.an.,V.o.Riquelme et al.,1996 haloplanktisŽ. Alteromonas sp.Palaemon macrodactylus Lagenidium callinectes Gil-Turnes et al.,1989Ž. Alteromonas sp.Pecten maximus Ps.d.,Pa.p.,Vibrio spp.Ruiz et al.,1996Ž. Alteromonas-like Pen.monodon hatchery Vibrio spp.Tanasomwang et al.,1998Ž. Bacillus sp.Callionymus sp.V.Õulnificus Sugita et al.,1998Ž. Carnobacterium Salmo salar V.an.,V.salmonicida Strøm,1988diÕergensŽ.¨Carnobacterium sp.Sa.salar Ae.s.,V.an.Joborn et al.,1997Ž. Lactococcus lactis Br.plicatilis V.anguillarum Shiri Harzevili et al.,1998Ž. Lactobacillus sp.Paralichthys oliÕaceus Ae.h.,Ed.t.,Pa.p.,V.an.Byun et al.,1997Ž. Pseudoalteromonas marine environment IHNV,V.an.Maeda et al.,1997 undinaŽ.Ps.fluorescens Lates niloticus V.anguillarum Gram et al.,1999Ž. Ps.fluorescens Salmo trutta Ae.salmonicida Smith and Davey,1993Ž. Roseobacter sp.Pec.maximus Ae.sp.,Ps.d.,Vibrio spp.Ruiz-Ponte et al.,1998Ž.T.utilis Pen.monodon Haliphthoros sp.Nogami et al.,1997Ž. T.utilis Pen.monodon V.anguillarum Nogami and Maeda,1992Ž. V.alginolyticus Pen.monodon Vibrio harÕeyi Ruangpan et al.,1998Ž.V.alginolyticus shrimp hatchery Ae.s.,V.an.,V.o.,Y.r.Austin et al.,1995Ž.Vibrio spp.marine environment V.parahaemolyticus Nair et al.,1985ŽVibrio spp.shrimp hatchery IHNV,OMV Direkbusarakom et al.,.1998Ž. Vibrionaceae Hippoglossus hippoglossus Vibrio sp.Bergh,1995()F.J.Gatesoupe r Aquaculture1801999147–165155agonistic strains are members of the Pseudomonas–Alteromonas and r or Vibrio groups Ž.Lemos et al.,1985;Nair et al.,1985.Antibacterial activity is also common inŽ.freshwater microbiota Sugita et al.,1996b.Some lactic acid bacteria,such as Ca.ŽdiÕergens and Lactobacillus sp.are antagonistic to fish pathogens Strøm,1988;Byun.Ž.¨et al.,1997;Joborn et al.,1997.Sugita et al.1998isolated a strain of Bacillus sp.that was antagonistic to63%of the isolates from fish intestine.Pathogenic strains of Vibrio or Aeromonas have been targeted in most in vitro tests,but some other fish pathogensŽwere also tested,e.g.,Ed.tarda,En.seriolicida,Pa.piscicida,Y.ruckeri Dopazo et al.,1988;Austin et al.,1995;Ruiz et al.,1996;Sugita et al.,1996a;Byun et al.,1997;.ŽGibson et al.,1998.Some bacteria are antagonistic to viruses Kamei et al.,1987,1988;.Direkbusarakom et al.,1998,and they may be efficient for the biocontrol of viral Ž.diseases Maeda et al.,1997.It is important to remind that antagonism may be mediated not only by antibiotics, but also by many other inhibitory substances,for example:organic acids,hydrogen Ž.Žperoxide reviewed by Ringøand Gatesoupe,1998,siderophores Gram and Mel-.chiorsen,1996.The inhibition due to such compounds is highly dependent on the experimental conditions,which are different in vitro and in vivo.Therefore,the expression of antagonism in vitro is not a sufficient criterion to select candidate Ž.probiotics Riquelme et al.,1997,nor is sufficient the absence of antagonism to rule the Ž.strains out Rico-Mora et al.,1998.5.4.Intestinal colonization and transienceThe colonization potential is another important criterion to characterize probiotics, but transient bacteria may be also efficient if the cells are introduced at high dose,Ž.continuously or semi-continuously Gournier-Chateau et al.,1994.In practice,it isŽ. therefore essential to evaluate the persistence of the probiotic in the gut Table2.TheŽexperimental introduction of lactic acid bacteria has been already reviewed Ringøand .Gatesoupe,1998.The concentration of Ca.diÕergens was higher in the pyloric caecaŽ.than in the intestine of Atlantic cod juveniles Gildberg and Mikkelsen,1998.Isolates of lactic acid bacteria seemed able to survive for several days in the intestine of larval Ž.¨and juvenile fish Strøm and Ringø,1993;Joborn et al.,1997.Vibrionaceae may alsoŽpersist for days or weeks in fish Austin et al.,1995;Munro et al.,1995;Ringøand .Ž. Vadstein,1998and in Pacific oyster larvae,Cr.gigas Gibson et al.,1998.YeastsŽ.seemed particularly persistent in rainbow trout Andlid et al.,1995.Adhesion to intestinal mucus was also assayed in vitro.Such tests indicated that Carnobacterium sp.adhered indifferently to the intestinal mucus of rainbow trout or toŽ.¨control surface treated with bovine serum albumin Joborn et al.,1997.Autochthonous intestinal bacteria of turbot seemed to adhere specifically to intestinal mucus,since their adhesion potential was stronger to mucus than to control surface,whereas bacteriaŽ.isolated from skin mucus were poorly adhesive Olsson et al.,1992.Yeasts also adhereŽ.to the intestinal mucus of rainbow trout Vazquez-Juarez et al.,1997,and theŽ. involvement of specific adhesins has been demonstrated Vazquez-Juarez,1996.Yeasts have therefore a great potential to adhere and to colonize the intestine of fish,and their application as probionts in aquaculture deserves more attention.。

Reovirus infection emerged in cultured channel cat fish,Ictalurus punctatus ,in ChinaJin Xu a ,b ,Lingbing Zeng b ,⁎,Xiaosong Luo b ,Yao Wang a ,Yuding Fan b ,Shiyuan Gong a ,⁎⁎aCollege of Fisheries,Huazhong Agricultural University,Wuhan 430070,PR Chinab Division of fish pathology,Yangtze River Fisheries Research Institute,Chinese Academy of Fishery Sciences,The Key Laboratory of Yangtze River Basin ’s Aquatic Animal Diseases,The Ministry of Agriculture of China,Wuhan 430223,PR Chinaa b s t r a c ta r t i c l e i n f o Article history:Received 18September 2012Received in revised form 8October 2012Accepted 8October 2012Available online 17October 2012Keywords:Channel cat fish (Ictalurus punctatus )Reovirus IsolationIdenti fication SequenceA reovirus,designated as CCRV-730,was isolated from channel cat fish,Ictalurus punctatus ,fingerlings suffering a severe hemorrhage in Hubei province in China.Experimental infection con firmed the pathogenicity of the virus and proved it to be the causative pathogen.Signs of the diseased channel cat fish included abdominal distention,eyes bulging and hemorrhages of operculum,lower jaw,skin and fin bases.The CCK cell line was used for viral pathogen isolation.The electron microscopy observation of the virus infected cells revealed that there were a large number of reovirus-like particles measuring 60–70nm in diameter in cytoplasm arrayed in crystalline.The viral genomic RNA was extracted and analyzed by SDS-PAGE and the complete S-class (S7–S11)segments of genome RNA were cloned and sequenced.The sequence alignment analysis of the S-class segments of CCRV-730with corresponding sequences of other Aquareovirus members in the genetic sequence database of NCBI indicated that the virus had high similarity to grass carp reovirus,especially shared 99%–100%nucleotide sequence identity with the grass carp reovirus 873strain (GCRV-873).The results implied that the genetic variation of GCRV-873potentially arose in natural environment and resulted in the viral host conversion or expansion and made it pathogenic to channel cat fish.©2012Elsevier B.V.All rights reserved.1.IntroductionChannel cat fish (Ictalurus punctatus )is the primary species of fish cultured in the southeast region of the United States.It was intro-duced into China in 1984and has become an important species farmed in nearly 20provinces in China.According to production estimates,the annual output of farmed channel cat fish in China was about 220,000metric tons in 2009(Deng et al.,2010).With the in-creasing production there has been more epizootic diseases detected in farmed channel cat fish in China because of high density stocking,genetic depression and improper fish health management (Ge et al.,2001).Thus,the diseases have become the major threat to the channel cat fish industry in China.Besides bacterial pathogens,viral pathogens are also considered as the primary threat because of the acuteness,high mortality and lack of effective treatment of disease.To date,only two viruses,channel cat fish virus (CCV;synonyms:Ictalurid herpesvirus -1,Herpesvirus ictaluri )(Fijan,1968)and channel cat fish reovirus (CRV)(Amend et al.,1984),have been isolated and identi fied in channel cat fish.Channel cat fish virus disease (CCVD)was considered as the most serious viral disease where channel cat fish were cultured in USA.The mortality could reach 100%in someproduction groups (Alvin and Camus,2004).On the other hand,CRV did not cause any notable economic losses in cultured cat fish in the USA (Hedrick et al.,1984).Although channel cat fish have been raised in China for many years,no virus had been isolated and identi fied before.Recently,a hemorrhagic disease in cultured channel cat fish finger-lings broke out and spread in the central part of China.The main pathological features were hemorrhages of operculum,lower jaw,skin,fin bases,abdominal distention,and eyes bulging.This disease caused up to 60%mortality in cultured channel cat fish fingerlings less than 12cm in length at a water temperature of 28°C.A reovirus was isolated and identi fied by cell culture and electron microscopy observations as well as molecular characterizations.This study had made a solid base for the diagnosis and control of the newly emerged viral disease in farmed channel cat fish.2.Materials and methods 2.1.FishMoribund channel cat fish,which were 6–12cm in length and had a typical hemorrhagic disease,were collected from a farm located in Zhijiang City,Hubei Province,China.Healthy channel cat fish fingerlings for experimental infection,with a length of 5–10cm,were collected from another farm that had no previous history of this disease.The fish were maintained in aerated tanks at a tempera-ture of 28°C for 1week before experimental infection.Aquaculture 372–375(2013)39–44⁎Corresponding author.Tel.:+862781780158.⁎⁎Corresponding author.E-mail addresses:zenglingbing@ (L.Zeng),gsy@ (S.Gong).0044-8486/$–see front matter ©2012Elsevier B.V.All rights reserved./10.1016/j.aquaculture.2012.10.011Contents lists available at SciVerse ScienceDirectAquaculturej ou r n a l h o m e p a g e :w w w.e l s e v i e r.c o m /l o c a t e /a q u a -o n l i n e2.2.Parasitology and bacteriologyThe exterior mucus,gills and viscera were taken for parasite examination through an optical microscope.For bacteria isolation, liver and kidney materials from eachfish were inoculated onto brain heart infusion(BHI;Difco,USA)agar plates and incubated at28°C for5days.Any colonies were identified by using Biolog Automated Microbial Identification System(Biolog,USA).2.3.Virus isolation and propagationThe channel catfish kidney cell line(CCK,CCTCC NO:C201198) (Zeng et al.,2009)was used for virus isolation and propagation. The viscera including the kidney and spleen of diseasedfish were collected and homogenized with Dulbecco's Phosphate-Buffered Saline(DPBS,Sigma,Germany)on ice,then the homogenate was centrifuged at4000rpm for30min at4°C,and the supernatant was filtrated through a0.22μmfilter(Nalgene,USA).Thefiltered superna-tant(1ml)was supplemented with10μl polybrene(10mg ml−1) and inoculated onto a24-hour-old cell monolayer in a25cm2flask (Corning,USA).The same volume of cell culture medium was used for a mock infection control in cells.After the adsorption for1h at 28°C and extraction of the inoculums,5ml of medium supplemented with2%FBS was added to theflasks,and then the infected cells were incubated at28°C in CO2incubator for7days.After incubation for7days at28°C,the CCK monolayers showing no CPE were subcultured by transferring0.5ml of culture medium supernatant onto fresh cell monolayers and incubated for a further7days.The infected cultures were observed daily under an inverted phase con-trast microscope(Nikon,Japan).When80%cytopathic effect(CPE) was observed the cultures were harvested and stored at−80°C for further identification.The continuous infection of the isolated virus in the CCK cell line was followed as above.The virus titer was calculated using the Spearman–Karber formula(Wardlaw,1985).2.4.Electron microscopyAfter CCRV-730was grown in CCK cells in a25cm2flask for36to 48h,the culture medium was aspirated out and the cells werefixed in2.5%(V/V)glutaraldehyde in a0.1M cacodylate buffer(pH7.3) for1h at4°C.Thefixed cells were scraped down by a cell scraper (Fisher,USA)and pelleted by centrifugation at2500rpm for 20min.The cell pellet was post-fixed in1%(W/V)cacodylic OsO4 for1h and dehydrated through a graded ethanol series(50–100%), then the cell pellets were impregnated at60°C for48h with epoxy resin.Ultra-thin sections(80nm)of the cell pellet were cut using a microtome(Leica UC7,Germany).After being double stained with uranyl acetate and lead citrate,the sections were examined and photographed by transmission electron microscopy(Hitachi-7650, Japan)at80kV.2.5.Virus genome RNA extraction and SDS-PAGE analysisThe CCRV-730genomic RNA was extracted from the virus prepa-ration obtained from both the diseasedfish and the infected CCK cells.Briefly,the tissue homogenate of the diseasedfish and the virus infected cells were frozen–thawed twice between−80°C and room temperature,and then the samples were centrifuged at 5000rpm at4°C for30min.The supernatant was ultra-centrifuged at25,000rpm at4°C for2h(SW28,Beckman-Coulter,Optima LX-80,USA).The pellet was suspended in DPBS and used for RNA extraction with TRIzol LS Reagent(Invitrogen,USA)according to the protocol provided.The RNA was dissolved in an appropriate volume of nuclease free water(Promega,USA)and resolved by SDS poly-acrylamide gel electrophoresis(SDS-PAGE)with a10%gel by the method of Laemmli(Laemmli,1970).After electrophoresis for20h at9mA,the RNA was visualized after staining with silver reagents (Merril et al.,1981).The size of each virus genome segment was esti-mated by comparing its relative mobility with a1kb double-stranded DNA ladder(Fermentas GeneRuler).2.6.Sequence determination of S-class segments of the virus genomeA modified method based on the single primer amplification technique(SPAT)described previously(Attoui et al.,2000;Qiu et al.,2001)was used for the virus genome sequence determination. Briefly,the virus genome RNA segments were separated by agarose gel electrophoresis,and the S-class segment RNA bands were excised and extracted by silica beads(Fermentas).Oligodeoxyribonucleotide primer A(5′-PO4-ATTTACCGCCGAGCCTGACTT-NH2-3′)(Qiu et al., 2001)that had been modified with5′-phosphagen and3′-amidogen to prevent self-ligation and subsequent concatenation,was ligated to both3′ends of the dsRNA using10U of T4RNA ligase I(NEB, USA).The ligation mixture was incubated at16°C for12h.Tailed RNA was recovered from the ligation mixture using silica beads. The primer one-tailed dsRNA was denatured by heating at94°C for5min in15%DMSO in the presence of 1.5mM primerB (5′-AAGTCAGGCTCGGCGGTAAAT-3′)complementary to primer A. The mixture was immediately cooled in an ice–water bath to prevent the re-annealing of the dsRNA.Synthesis of cDNA was carried out at 55°C for1h in a reverse transcriptase reaction mix in a total volume of20μl containing:10μl of the DMSO-denatured dsRNA solution (final DMSO concentration in the mixture is7.5%),1×cDNA synthesis buffer,5mM DTT,0.2mM each dNTP,40U of Ribonuclease inhibitor (Promega,USA)and15U of ThermoScript™reverse transcriptase (Invitrogen,USA).The amplification of cDNA was accomplished by PCR with only primer B that consisted of a denaturation step at 94°C for3min followed by40cycles of45s at94°C,45s at56°C, and90s at72°C.After the amplified DNA products were separated on an agarose gel,the fragments were excised and purified.These were directly ligated into pGEM-T easy vector(Promega,USA) and transformed into a competent DH5αstrain of Escherichia coli (Invitrogen,USA).The recombinant plasmids containing the full-length cDNA of the genome segments were identified according to the size of the inserted segment by PCR using T7and SP6primers on pGEM-T easy.Positive clones that had the full-length cDNA were sequenced in different directions.The sequencing was done by a bio-technology company(Sangon,China).Nucleotide and amino acid sequences were compiled and ana-lyzed with the programs included in the DNASTAR software package (Lasergene).Putative open reading frames(ORFs)were predicted byfinding the start codon AUG and the rest of the coding sequence with the DNASTAR software package.The DNA and protein compari-sons with entries in the sequence databases were performed with the BLAST tool in NCBI.2.7.Experimental infection and RT-PCR assayExperimental infections were performed in healthy channel catfish(5–10cm in length)with the homogenatefiltrate of tissues of the naturally infected channel catfish and the cell cultured virus. Two groups of30channel catfish were injected intraperitoneally with0.1mlfiltrate and cell cultured virus(passage5,106TCID50fish−1)respectively.Another group of30channel catfish were injected with0.1ml PBS for mock-infection.Allfish of the three groups were kept in aerated tanks at a temperature of28°C.Clinical signs and mortality were observed daily.Three moribund or deadfish in each infection group and three livefish in mock-infection group were randomly selected for virus detection by RT-PCR assay.A set of PCR primers was designed based on S8gene sequence for CCRV-730detection.A327bp gene fragment was amplified by RT-PCR using oligonucleotide primers PF(5′-GGCGAGACCCAGTCCTAT-3′)and40J.Xu et al./Aquaculture372–375(2013)39–44PR (5′-CGGGTGATTGTGGCAGTT-3′).The spleen,liver and kidney tissues of the experimental fish were collected and homogenized for viral RNA template extraction according to the method described above.The RT-PCR reaction was carried out using TITANIUM ™One-Step RT-PCR Kit (Clontech,USA)according to the manufacturer's recommended instructions.Ampli fication products were analyzed by electrophoresis through 1.5%agarose gels and ethidium bromide staining.3.Results3.1.Outbreaks and gross pathological signsThe diseased channel cat fish fingerlings (6–12cm)presented lethargic,separately floating at the surface,spiral swimming move-ments and finally died within 24h,showing severe hemorrhage at operculum,lower jaw and fin bases,and body color depigmentation,bilateral exophthalmia,and abdominal distention (Fig.1).On post-mortem examination,gills of diseased fish presented blood loss anemia and become pale.The liver was pale and contained small hemorrhages,and the spleen was dark and enlarged.The digestive tract contained no food but hemorrhages on the intestinal wall.Clear yellow to blood-tinged fluid was presented in the body cavities of abdominal distention individuals.3.2.Parasitology and bacteriologyParasitology inspection showed no notable parasite appeared in the exterior mucus,gills and viscera.Bacteria were also not consis-tently isolated from diseased fish.3.3.Virus isolation and propagationThe tissue homogenate filtrate obtained from the viscera of diseased channel cat fish was used for the virus isolation in the CCK cell line.Cytopathic effect was observed by light microscopy at 48h post-inoculation (Fig.2).The first sign of CPE was the shrinking of cells and a small black spot that appeared in the cytoplasm of these cells.Additionally,other cells became round and lost their typical fibroblastic shape.This was followed by vacuolation and the cells detaching from the monolayer and lysing.The CPE was most promi-nent at 96h post-inoculation with >90%of the cells exhibiting somesort of abnormal change.The mock-infected CCK cells exhibited no CPE at 96h post-inoculation.The resultant virus titer was calculated to be 4.64×108TCID 50/ml.3.4.Electron microscopyIn the CCK cells,CCRV-730particles had icosahedral symmetry with double capsid structure.The virions in cell culture had an overall diameter of about 60–70nm (Fig.3).Characteristic cytoplasmic in-clusion bodies of variable size and shapes with relatively electron dense areas of granular matrix were found scattered in the cytoplasm without any special membranous envelope at different stages ofvirusFig.1.The signs of diseased channel cat fish fingerlings,showing severe hemorrhage at operculum,fins and fin bases,a swollen abdomen and bulgingeyes.Fig.2.Cytopathic effect (CPE)in the channel cat fish kidney (CCK)cells.A:mock-infected CCK cells.B:CPE in the CCK cells 48h after virus infection.Bar=100μm.Fig.3.Electron microscopy observation of CCRV-730in the infected CCK cells.Arrow A:crystalline-arrayed virus particles,with a diameter of about 60–70nm,bar=200nm;B:cell nucleus (collapsed);and C:cell membrane.41J.Xu et al./Aquaculture 372–375(2013)39–44formation.Some of the viroplasms were found to be fully formed with a crystalline array of virus particles filling the entire area of the inclusion body.Morphologically,CCRV-730was similar to the other aquareoviruses such as grass carp reovirus (GCRV)and golden shiner reovirus (GSRV).3.5.Virus genome RNA extraction and SDS-PAGE analysisThe virus genome RNA extracted from both the diseased fish tissues and the virus infected CCK cells showed the same electrophoretype in SDS-PAGE (Fig.4).SDS-PAGE revealed that the virus had a genome composed of 11segments,a typical electrophoretype of the Reoviridae genome.The segments fell into three size classes that ranged from 0.9kb to 4.4kb compared with the dsDNA ladder.The 11segments completely separated and formed a unique electrophoretype,three large,three medium and five small bands.3.6.Sequence determination of S-class segments of the virus genome The complete nucleotide sequence of S-class segments including S7,S8,S9,S10and S11was determined.All the sequences were depos-ited in GenBank (GenBank accession nos.HM545301,HM545302,HM545303,HM545304and HM545305).The RNA segment length,the longest ORF,the sizes of the 5′and 3′non-coding regions (NCR)and the size of the putative protein were identi fied for each segment and are shown in Table 1.The S7segment was 1414bp in length.The ORF analysis showed that S7was bicistronic,encoding two proteins.The first protein consisted of 146aa (15,775Da)and was encoded by the sequence be-tween bases 14and 454.The second protein was 274aa (31,214Da)and was encoded by the sequence between bases 519and 1344.The total length of S8was 1296bp and the ORF sequence was located between bases 12and 1250that encoded a 412aa protein (44,589Da).S9was 1130bp in length and encoded a 352aa protein (37,693Da).The ORF sequence in S9was located between bases 32and 1090.S10was 909bp and encoded a 276aa protein (29,851Da).The ORF sequence on S10was located between bases 31and 861.S11was 820bp containing an ORF between bases 43and 777and encoding a 244aa protein (26,417Da).Segments S7–S11of the virus were found to have conserved ter-minal sequences.All RNA segments had the motif 5′G/UUUAUUU/G3′in common at the 5′end and the motif 5′UUCAU/GC3′in common at the 3′end.The function of the conserved motifs is thought to act as sorting signals,bringing a single copy of each genome segment into the nascent virus capsid (Anzola et al.,1987).As Table 2showed,the blast comparison of the S-class segments of CCRV-730with other Aquareovirus in the NCBI nucleotide database showed that they had high similarity to the same segments of the grass carp reovirus strains (GCRV:90–100%in identity)and golden shiner reovirus (GSRV:91–97%in identity),and had a relatively low similarity to the American grass carp reovirus (AGCRV,strain PB01-155:66–85%in identity).3.7.Experimental infectionChannel cat fish inoculated with 0.1ml tissue homogenate filtrate began to die at 2day post-inoculation (dpi),and the death rate reached 50%at 7dpi (Fig.5A).In the other infection group which in-oculated with 106TCID 50fish −1cell cultured virus,the first death had appeared at 1dpi and finally reached 83.3%at 12dpi.Both groups went through a 3–4days rapid death stage.The dead fish of infection groups presented the same hemorrhagic signs as those of natural infected fish.There was no mortality in mock-infection group.The RT-PCR detection of the dead fish challenged with both tissue homogenate filtrate and cell cultured CCRV-730was all positive,while that of the live fish of mock-infection group was detected negative (Fig.5B).The results con firmed the pathogenicity of the virus and proved CCRV-730to be the causative pathogen of the channel cat fish hemorrhagedisease.Fig.4.SDS-PAGE electrophoretype of the CCRV-730genome RNA segments.M:DNA ladder;A:CCRV-730genome RNA extracted from diseased fish tissues;and B:CCRV-730genome RNA extracted from the CCK culture.Table 1Properties of S-class segments of CCRV-730including the RNA segment length,the amino acid length and mass of the putative protein encoded,the sizes of the 5′and 3′non-coding regions and the GenBank accession number.Segment nameNucleotide length (nt)Putative encoded protein 5′NCR 3′NCR Accession numberLength (aa)Mass (Da)Length (nt)Terminal sequence Terminal sequence Length (nt)S7*******,27415,775,31,214135′UUUAUUU UUCAGC3′70HM545301S8129641244,589115′GUUAUUU UUCAUC3′46HM545302S9113035237,693315′GUUAUUU AUCAUC3′40HM545303S1090927629,851305′GUUAUUU UUCAUC3′48HM545304S1182024426,417425′GUUAUUGUUCAUC3′43HM54530542J.Xu et al./Aquaculture 372–375(2013)39–444.DiscussionAlthough channel catfish has been cultured for28years in China, there was no confirmed viral disease reported yet.In2005,Geng and Wang reported a suspected viral disease in cultured channel catfish in Sichuan Province,based on pathological observation of the diseased channel catfish tissue sections and ultrathin sections by light and electron microscopic examination,they draw a conclusion that it was a herpesvirus(Geng and Wang,2005),but no more direct evidences such as isolation and cell culture of virus,and clear mor-phological observation of virus particles presented.To date,the chan-nel catfish hemorrhagic disease caused by reovirus was the only viral disease of cultured channel catfish reported in China.From the clinical symptoms,most of the diseasedfish showed typical severe hemorrhagic symptoms,but a small part of the diseased channel catfish presented almost the same symptoms as those of CCVD such as abdominal distention and eyes bulging(Alvin and Camus,2004).These signs initially lead us to speculate that it was CCVD,but we failed to detect the CCV in diseasefish tissue according to the PCR detection method by Gray(Gray et al.,1999). Moreover,CCV particles were not observed in the infected CCK cell ultrathin sections by EM.These negative results confirmed the absence of CCV in this disease.The results of experimental infection confirmed the pathogenicity of the virus and proved CCRV-730to be the causative pathogen of the channel catfish hemorrhage disease.CCRV-730could propagate in the CCK cells,caused typical cyto-pathic effect(CPE)and reached high viral titer.The electron micros-copy observation revealed that the virus replicated in cytoplasm, arrayed in crystalline,and had a non-enveloped double capsid with a diameter of60–70nm.The SDS-PAGE analysis of the virus genome RNA demonstrated that CCRV-730was a reovirus belonged to Reoviridae pared with the channel catfish reovirus(Hedrick et al., 1984)isolated in USA on the SDS-PAGE electrophoretype and the rele-vant segment size,CCRV-730was distinct to CRV.Sequence determination of S-class segments of CCRV-730showed that it had a very high similarity to GCRV and especially to the GCRV-873stain.Based on the high identity of the S-class segment sequences between this two virus strains,we could speculate that CCRV-730may be derived from GCRV-873despite the fact that CCRV-730was isolated from channel catfish.Grass carp reovirus is the causative agent of grass carp hemor-rhagic disease in China,which causes serious economic loss in the grass carp industry in China(Fang and Zhu,2003).The grass carp hemorrhage wasfirst observed in1953(Ni,1994)and the pathogen wasfirst isolated in1983in cell culture(Chen and Jiang,1983).Pre-vious results showed that,in addition to grass carp,GCRV could infect black carp(Mylopharyngodon piceus)and rare gudgeon(Gobiocypris rarus)and cause the same hemorrhagic disease.Additionally,GCRV could be detected in silver carp(Hypophthalmichthys molitrix)and Hemiculter bleekeri but could not cause visible disease(Ding et al., 1991).These fourfish species belong to the same Cyprinidae family as grass carp.However,other species within the Cyprinidae family such as common carp(Cyprinus carpio),crucian carp(Carassius auratus),big-head carp(Aristichthys nobilis)and bluntnose black bream(Megalobrama amblycephala)did not show any susceptibility to GCRV(Ding et al.,1991).It was concluded that GCRV had a high specificity for specificfish in Cyprinidae family.Hence it is difficult to believe that GCRV could infect channel catfish and cause a similar hemorrhagic disease.The GCRV-873strain was isolated from diseased grass carp cul-tured in Shaoyang City,Hunan province in1987(Ke et al.,1990).In a previous study on GCRV-873,this virus did not replicate in a chan-nel catfish ovary cell line(CCO)(Zhang et al.,2003).This suggests that GCRV-873could not infect channel catfish.Based on our results, we had reason to suspect that,some genetic variation of GCRV-873, even a minor change,may potentially arise in natural environment and result in this“novel”virus expanding its hosts and becoming pathogenic to channel catfish.The changed part of the genome needs to befigured out by a complete genome sequencing and comparative analysis to explain the host conversion or expansion of GCRC-873strain.Theσ1protein of the mammalian reovirus(MRV) played the most important role in thefirst step of virus infection (Mercier et al.,2004;Montufar-Solis and Klein,2005).This protein was important for virus attachment on the host cell receptor on the cell surface.But the3D reconstruction of the GCRV surface showed that GCRV did not have the same structure asσ1protein(Fang et al.,Table2The comparison of S-class segment genome similarity between CCRV-730and other stains of Aquareovirus including grass carp reovirus(GCRV),golden shiner reovirus(GSRV)and American grass carp reovirus(AGCRV).CCRV-730S7CCRV-730S8CCRV-730S9CCRV-730S10CCRV-730S11Query coverage Max ident Query coverage Max ident Query coverage Max ident Query coverage Max ident Query coverage Max identGCRV-87399%99%100%99%100%99%100%99%100%100% GSRV99%95%100%94%100%92%100%91%100%97% GCRV-876–a–85%100%––84%99%––GCRV-875––85%100%––84%90%––GCRV-991––85%99%––84%99%––AGCRV2%85%68%66%63%72%0000a“–”means no comparison result because of the lack of nucleotide information of relevant GCRVstrain.Fig.5.Pathogenicity of the CCRV-730isolated from the natural infected channel cat-fish.A:cumulative mortality of the CCRV-730infected channel catfish.B:CCRV-730 RT-PCR detection of thefish in experimental infection.M:DL1000DNA marker;lane 1:positive control RNA template from the natural infected channel catfish;lane2–4: 3individual deadfish challenged with tissue homogenatefiltrate of the natural infected channel catfish;lane5–7:3individual deadfish challenged with cell cultured CCRV-730;and lane8–10:3individual livefish from mock-infection group.43J.Xu et al./Aquaculture372–375(2013)39–442005).It indicated that the virus invasion mechanism of GCRV was different from that of MRV.The relationship and difference between CCRV-730and GCRV-873need to be further determined,and it may help us in understanding the mechanism of virus infection of the aquareoviruses.AcknowledgmentsThis study was supported by the Earmarked Fund for China Agriculture Research System(CARS-46)and the Special Fund for Agro-scientific Research in the Public Interest(200803013). ReferencesAlvin,C.,Camus,2004.Channel catfish virus disease.SRAC Publication No.4702. Amend,D.,McDowell,T.,Hedrick,R.,1984.Characteristics of a previously unidentified virus from channel catfish(Ictalurus punctatus).Canadian Journal of Fisheries and Aquatic Sciences41,807–811.Anzola,J.,Xu,Z.,Asamizu,T.,Nuss,D.,1987.Segment-specific inverted repeats found adjacent to conserved terminal sequences in wound tumor virus genome and defective interfering RNAs.Proceedings of the National Academy of Sciences of the United States of America84,8301.Attoui,H.,Billoir,F.,Cantaloube,J.,Biagini,P.,de Micco,P.,de Lamballerie,X.,2000.Strategies for the sequence determination of viral dsRNA genomes.Journal of Virological Methods89,147–158.Chen,Y.X.,Jiang,Y.L.,1983.Morphological and physio-chemical characterization of the hemorrhagic virus of grass carp.Kexue Tongbao28,1138–1140.Deng,W.,Xiao,Y.,Cao,K.,Ma,L.,2010.Development,challenge and strategy of channel cafish industry of China.China Fisheries12,11–13.Ding,Q.,Yu,L.,Ke,L.,Cai,Y.,1991.Study on infecting otherfishes with grass carp hemorrhage virus.Virologica Sinica6,371–373.Fang,Q.,Zhu,Z.,2003.Advances in Aquareovirus research.Virologica Sinica18,82–85. Fang,Q.,Shah,S.,Liang,Y.,Zhou,Z.H.,2005.3D reconstruction and capsid protein characterization of grass carp reovirus.Science in China.Series C,Life Sciences48, 593–600.Fijan,N.,1968.Progress report on acute mortality of channel catfishfingerlings caused by a virus.Bulletin-Office International Epizooties69,1167–1168.Ge,L.,Ge,H.,Zhang,Y.,2001.The culture technology and disease control of channel catfish.Reservoir Fisheries21,48–50.Geng,Y.,Wang,K.,2005.Pathomorphologic observation of channel catfish Ictalunes punctatus infected with herpesvirus.Chinese Journal of Veterinary Science25, 636–639.Gray,W.L.,Williams,R.J.,Jordan,R.L.,Griffin,B.R.,1999.Detection of channel catfish virus DNA in latently infected catfish.Journal of General Virology80(Pt7),1817–1822. Hedrick,R.,Rosemark,R.,Aronstein,D.,Winton,J.,McDowell,T.,Amend,D.,1984.Characteristics of a new reovirus from channel catfish(Ictalurus punctatus).Journal of General Virology65,1527.Ke,L.,Fang,Q.,Cai,Y.,1990.Characteristics of a novel isolate of grass carp hemorrhagic virus.Acta Hydrobiologica Sinica14,153–158.Laemmli,U.,1970.Cleavage of structural proteins during the assembly of the head of bacteriophage T4.Nature227,680–685.Mercier,G.,Campbell,J.,Chappell,J.,Stehle,T.,Dermody,T.,Barry,M.,2004.A chimeric adenovirus vector encoding reovirus attachment proteinσ1targets cells expressing junctional adhesion molecule.Proceedings of the National Academy of Sciences of the United States of America101,6188.Merril,C.,Goldman,D.,Sedman,S.,Ebert,M.,1981.Ultrasensitive stain for proteins in polyacrylamide gels shows regional variation in cerebrospinalfluid proteins.Science(New York,NY)211,1437.Montufar-Solis,D.,Klein,J.,2005.Experimental intestinal reovirus infection of mice.Immunologic Research33,257–265.Ni,D.,1994.Recent developments and prospects offish disease science in China.Journal of Modern Fisheries Information9,1–4.Qiu,T.,Lu,R.H.,Zhang,J.,Zhu,Z.Y.,2001.The molecular characterization of RNA segment S9of grass carp hemorrhage virus(GCHV),an Aquareovirus.Aquaculture 203,1–7.Wardlaw,A.C.,1985.Practical Statistics for Experimental Biologists.Wiley,Chichester, New York.x,290p.pp.Zeng,L.B.,Li,X.L.,Zhang,L.,Xu,J.,Zhang,Y.,2009.Establishment and characterization of a cell line derived from kidney of channel catfish,Ictalurus punctatus.Journal of Fishery Sciences of China16,75–81.Zhang,Q.,Ruan,H.,Li,Z.,Zhang,J.,Gui,J.,2003.Detection of grass carp hemorrhage virus(GCHV)from Vietnam and comparison with GCHV strain from China.High Technology Letters9,7–13.44J.Xu et al./Aquaculture372–375(2013)39–44。

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