Vaccine

Abstract

The present invention is concerned with the development of a vaccine against Aeromonas hydrophila for use especially in fish. The invention provides an immunogenic S-layer protein of approximately 50 kDa of A. hydrophila for use in the development of a vaccine, as well as the nucleic acid encoding said protein and vaccines comprising said protein or nucleic acid encoding said protein.

1 . A method for immunising, vaccinating or raising an immune response in freshwater fish or shellfish as against  A. hydrophila  by administering to the fish or shellfish a formulation comprising:
 a polynucleotide fragment having a nucleotide sequence of SEQ ID NO: 1 or 3 that encodes a polypeptide having an amino acid sequence of SEQ ID NO: 2 or 4 respectively, an antigenic analog, fragment, or modified version thereof; or   a polypeptide having an amino acid sequence of SEQ ID NO: 2 or 4, an antigenic analog, fragment, or modified version thereof   
     
     
         2 . The method according to  claim 1  wherein the formulation further comprises an adjuvant. 
     
     
         3 . The method according to  claim 1  wherein the polynucleotide or polypeptide is packaged within a micro-particulate delivery system, selected from the group consisting of latex beads, poly(lactide-co-glycolide) microspheres, atelocollagen “minipellets”, bentonite, orporous apatite ceramics including hypoxyapatite (HA) and beta-tricalcium phosphate (TCP). 
     
     
         4 . The method according to  claim 1  for immunising, vaccinating or raising an immune response in Abalone  Haliotis discus hannai ; Adriatic sturgeon  Acipenser naccarii ; African catfish  Clarias bartachus ; African cichlid  Nimbochromis venustus ; Amberjack  Seriola dumerili ; American crayfish  Orconectes limosus, Pacifasacus leniusculus, Procambarus clarkia ; American plaice  Hippoglossoides platessoides;  Arctic char  Salvelinus alpinus  L.; Atlantic cod  Gadus morhua  L.; Atlantic croaker  Micropogon undulates ; Atlantic menhaden  Brevoortia tyrannus  Latrobe; Atlantic salmon  Salmo salar  L.; Australian oyster  Saccostrea commercialis ; Ayu  Plecoglossus altivelis ; Balloon molly  Poecilia , spp.: Banana prawn  Penaeus merguiensis ; Barramundi  Lates calcarifer  (Bloch); Bighead carp  Aristichthys nobilis ; Black acara  Cichlasoma bimaculatum ; Black mullet  Mugil cephalus ; Black scraper  Novodon modestus ; Black skirted tetra  Hyphessobrycon  spp.; Blenny  Zoarces viviparous ; Blue fish  Pomatomus saltatrix ; Blue mackerel  Scomber australasicus ; Blue manna crab  Portunus pelagicus ; Blue shrimp  P . ( Litopenaeus )  stylirostris ; Boney bream  Nematolosa come  (Richardson); Borneo mullet  Liza macrolepis ; Bream  Abramis brama ; Brine shrimp  Artemia ; Brook salmon  Salvelinus fontinalis ; Brook trout  Salvelinus fontinalis  (Mitchill); Brown bullhead  Ictalurus nebulosus  (Lesueur); Brown trout  Salmo trutta ; Brown-spotted grouper  Epinephelus tauvina, E. Coioides ; Bullfrog  Rana catesbeiana ; Burnett salmon  Polydactylus sheridani  (Macleay); Canadian shrimp  Lismata amboiens ; Carp  Cyprinus carpio  L.; Catfish  Clarius batrachus  L.; Caucasian carp  Carassius carassius ; Chanchito  Chichlasoma facetum  (Jenyns); Channel catfish  Ictalurus punctatus  (Rafinesque); Chinook salmon  Oncorhynchus tschawytscha ; Chub  Leuciscus cephalis ; Chum salmon  Oncorhynchus keta  (Walbaum); Cichlid  Oreochromis mossambicus ; Clam  Tapes philippinarum ; Cod  Gadus morhua ; Coho salmon  Oncorhynchus kisutch ; Common carp  Cyprinus carpio  L.; Common snook  Centropomus undecimalis ; Common wolf-fish  Anarhichas lupus ; Coral prawn  Metapenaeopsis  spp.; Crucian carp  Carassius carassius ; Cutthroat trout  Salmo clarki ; Cuttle fish  Sepia officinalis ; Cuvier Plaice  Pleuronectes platessa ; Dab  Limanda limanda ; Dace  Leuciscus leuciscus  L.; Damselfish  Chromis punctipinnis; Pomacentridae, Amphiprion clarkii  (Bennett),  Amblyglyphidodon  curacao (Bloch); Danio  Danio devario ; Discus fish  Symphysodon discus, S. aequifasciatus;  Dolphin fish  Coryphaena hippurus  L.; Eastern freshwater cod  Maccullochella ikei;  Eastern mosquitofish  Gambusia holbrooki  J; Eastern painted turtle  Chrysemys pieta pieta ; Eel  Anguilla japonica, A. reinhardtii; A. anguilla, A. Rostrata ; European crayfish  Astacus leptodactlus, A. pachypus, A. torrentium, A. astacus, Austropotamobius pallipes ; European sea bass  Dicentrarchus labra ; Fairy shrimp  Branchipus schaefferi  (Fisher),  Chirocephalus diaphanus  (Prevost),  Streptocephalus torvicornis  (Waga); Farmed mussel  Perna perna; Fathead minnow Pimephalespromelas ; Flat-tailed mullet  Liza dussumieri  (Valenciennes); Firemouth cichlid  Cichlasoma meeki ; Flounder  Paralichthys olivaceus, Platichthys flesus ; Four bearded rockling  Enchelyopus cimbrius  L.; Freshwater cod (Australian native)  Maccullochella  spp.; Freshwater prawn  Macrobranchium rosenbergii ; Golden shiner  Notemigonus crysoleucas  (Mitchell); Goldfish  Carassius auratus  L; Goldsinny wrasse  Ctenolabrus rupestris ; Gourami (three-spot)  Trichogaster trichopterus ; Grass carp  Ctenopharyngodon idella ; Grayling  Thymallus thymallus  L; Greater weever  Trachinus draco ; Green frog  Rana clamitans ; Green knife fish  Eigemannia virescens ; Green moray eel  Gymnothorax funebris ; Green sturgeon  Acipenser medirostris ; Greenback flounder  Rhombosolea tapirina ; Grouper  Epinephelus guaz, E. coioides ; Guppy  Poecilia reticulata  (Peters),  Lebistes reticulates ; Haddock  Melanogrammus aeglefinus  L.; Halibut  Hippoglossus hippoglossus ; L Herring  Arripis georgianus ; Horse mackerel  Trachurus trachurus ; Iberian toothcarp  Aphanius iberus ; Japanese abalone  Sulculus diversicolor supratexta ; Japanese medaka  Oryzias latipes ; Jewel tetra  Hyphessobrycon callistus  (Boulenger); King prawn  Penaeus latisulcatus ; Knife fish  Gymnotus carapo ; Lake trout  Salmo trutta  m  lacustris, Salvelinus namaycush  Walbaum; Largemouth bass  Micropterus salmoides ; Leopard frog  Rana pipiens ; Living dace  Tribolodon hakonensis  Gunther; Loach  Misgurnus anguil licaudatus  Cantor; Lobster  Homarus gammarus  L; Local mussel  Mutilus edulis  Long-tom  Tylosurus macleayanus  (Ogilby); Mackerel  Scomberscombrus ; Manila clam  Tapes philippinarum, T. decussatus, Ruditapes philippinarum ; Masu salmon  Oncorhynchus masou ; Menhaden  Brevoortia patronus ; Minnow  Phoxinus phoxinus  L.; Molly  Poecilia velifera  (Regan); Mud crab  Scylla serrata ; Mullet  Mugil cephalus;  Murray cod  Maccullochella peeli ; Mussel  Protothaca jedoensis, Mytilus edulis, Mytilus galloprovincialis ; Mussel (Far-eastern) Crenomytilus grayanus, Patinopecten yessoensis;  Neon tetra  Paracheirodon innesi, Hyphessobrycon innesi ; North-east Atlantic mackerel  Scomber scombrus ; Northern pike  Esox lucius  L.; Octopus  Octopus vulgaris, O. joubini;  One-spot bream  Diplodus sargus ; Ornamental fish  Pterophyllum scalare ; Oscar  Astronotus ocellatus; Apistogramma ocellatus ; Oyster  Ostrea edulis; Crassostreae virginica ; Pacific herring  Clupea harengus palla˜i;′  Pacific oyster  Crassostrea gigas ; Pacific salmon  Oncorhynchus  spp.; Pacific staghorn sculpin  Leptocottus armatus ; Pacific white shrimp  Penaeus vannamei ; Paradise fish  Macropodus opercularis  (L.); Pejerrey  Odonthestes bonariensis ; Perch  Perca fluviatilis ; Pickerel frog  Rana palustris ; Pike  Esox lucius ; Pilchard  Sardinops neopilchardu ; Pink salmon  Oncorhynchus gorbuscha ; Pink snapper  Chrysophrys unicolor ; Pinkfish  Lagodon rhomboids ; Pirarucu  Arapaima gigas ; Pompanos  Trachinotus carolinus  L; Rabbitfish  Siganus rivulatus  (ForsskAl); Rainbow trout  Oncorhynchus mykiss  (Walbaum); Red abalone  Haliotis ruftscens ; Red algae  Jainia  spp.; Red claw crayfish  Cherax quadricarinatus  L.; Red drum, Redfish  Sciaenops ocellatus ; Red sea bream  Pagrus major ; Red swamp crawfish  Procambarus clarkia ; Red-eared slider turtle  Chrysemys scripta elegans ; Redtail catfish  Phractocephalus hemiliopterus ; Regan Signal crayfish  Pacifastacus leniusculus ; Roach  Rutilus rutilus  L.; Rohu  Labeo rohita ; Rosy barb  Puntius conchonius ; Rudd  Scardinius erythrophthalmus;  Sablefish  Anoplopoma fimbria  (Pallas); Saltwater crocodile  Crocodylus porosus ; Sand eel  Ammodytes lancea  (Cuvier),  Hyperoplus lanceolatus  (Lesauvege); Sand lance  Ammodytes personatus  Girard; Sand whiting  Sillago ciliata  Cuvier;  Saratoga Scleropages leichardii ; Sardine  Sardinops melanostictus, Sardinops sagnax ; Scallop  Pecten maximus, Argopecten purpuratus ; Scaly mackerel fish  Amblygaster postera ; Sea bream  Pagrus major, Evynnisjaponicus, Sparus aurata, Acanthopagrus latus ; Sea catfish  Arius felis;  Sea horse  Hippocampus angustus, H. barbouri, H. whitei, H. kuda ; Sea mullet  Mugil cephalus  L.; Sea trout  Salmo trutta  m.  trutta  L.; Sea turtle  Chelonia mydas ; Sea-urchin  Paracentrotus lividu ; Sepiolid squid  Euprymna scolopes ; Shorted halibut  Eopsetta grigorjewi ; Shubunkin  Carassius  spp.; Siamese fighting fish  Betta splendens ; Silver black porgy  Acanthopagrus cuvieri, Pyoscelis papua ; Silver bream  Acanthopagrus butcheri, A. australis  (Owen); Silver bream  Blicca bjoerkna ; Silver carp  Hypophthalmichthys molitrix  Valenciennes; Silver molly  Poecilia  spp.; Silver mullet  Mugil curema  Valenciennes,  Mugil cephalus  L.; Silver perch  Bidyanus bidyanus  (Mitchell); Silver trout  Cynoscion nothus ; Small abalone  Haliotis diversicolor  supertexta; Smallmouth bass  Micropterus dolomieui ; Snakehead fish  Channa striatus; Ophicephalus punctatus, O. striatus ; Snub-nose garfish  Arrhamphus sclerolepsis  (Gunther); Sockeye salmon  Oncorhynchus nerka  (Walbaum); Softshell clam  Mya arenaria ; Sole  Solea solea ; South African abalone  Haliotis midae ; South American side-necked turtle  Podocnemis unifelis ; Spanish mackerel  Scomber japonicus ; Spanner crab  Ranina ranina ; Spiny soft-shelled turtle  Trionyx spinifer ; Spot  Leiostomus xanthurus;  Spotted moray eel  Gymnothorax moringa ; Spotted wolf-fish  Anarhichas minor ; Squid  Loligo pealei, Sepiola, T euthoidea ; Starfish  Asterias rubens ; Stingray  Dasyatis pastinaca ; Striped bass  Morone saxatilis  (Walbaum),  M. chrysops ; Striped mullet  Mugil cephalus ; Striped-neck musk turtle  Sternotherus minor  peltifer; Sturgeon  Acipenser naccari ; Sunfish  Mola mola ; Tasmanian lobster  Jasus novaehollandiae ; Tiger Frog  Rana tigrina; Tilapia Oreochromis niloticus, O. aurus; Sarotherodon aureus  (Steindachner);  Tilapia nilotica, Tilapia aurea, Tilapia mosambica ; Tom cod  Gadus microgadus;  Tropical shrimp  Stenopus hispidus ; Trout cod  Maccullochella macquariensis ; Turbot  Scophthalmus maximus  L.; Turtle  Dermochelys coriacea; Pseudemis scripta;  Viviparous blenny  Zoarces viviparus ; Western rock lobster  Panulirus cygnus ; White catfish  Ictalurus catus  L.; White clawed crayfish  Austropotamobius pallipes ; White leg shrimp  P . ( Litopenaeus )  vannamei ; White perch  Roccus americanus, Morone americanus  (Gremlin);  Whitefish Coregonus  spp.; Whitespotted rabbitfish  Siganus canaliculatus ; Whiting  Merlangius merlangus ; Wrasse  Labrus berggylta ; Yellow bass  Morone mississippiensis ; Yellow perch  Perca flavescens ; Yellowfin bream  Acanthopagrus australis  (Owen); Yellowtail  Seriola quinqueradiata, S. lalandi ; Zealand mussel  Perna canaliculus ; Zebra danio  Brachydanio rerio.    
     
     
         5 . The method according to  claim 1  wherein the polypeptide is part of a fusion protein.