NO346319B1 - Method for treating and/or preventing a sea lice infection in a teleost - Google Patents

Method for treating and/or preventing a sea lice infection in a teleost Download PDF

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Publication number
NO346319B1
NO346319B1 NO20191194A NO20191194A NO346319B1 NO 346319 B1 NO346319 B1 NO 346319B1 NO 20191194 A NO20191194 A NO 20191194A NO 20191194 A NO20191194 A NO 20191194A NO 346319 B1 NO346319 B1 NO 346319B1
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caligus
probiotic
probiotic composition
teleost
bacteria
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NO20191194A
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Norwegian (no)
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NO20191194A1 (en
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Kira Salonius
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Previwo As
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Priority to NO20191194A priority Critical patent/NO346319B1/en
Priority to EP20792930.8A priority patent/EP4037698B1/en
Priority to PCT/EP2020/077734 priority patent/WO2021064217A1/en
Priority to AU2020357769A priority patent/AU2020357769A1/en
Priority to CA3156807A priority patent/CA3156807A1/en
Priority to US17/766,551 priority patent/US20230338437A1/en
Publication of NO20191194A1 publication Critical patent/NO20191194A1/en
Priority to CL2022000819A priority patent/CL2022000819A1/en
Priority to DKPA202200420A priority patent/DK202200420A1/en
Publication of NO346319B1 publication Critical patent/NO346319B1/en

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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66Microorganisms or materials therefrom
    • A61K35/74Bacteria
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01KANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K61/00Culture of aquatic animals
    • A01K61/10Culture of aquatic animals of fish
    • A01K61/13Prevention or treatment of fish diseases
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K10/00Animal feeding-stuffs
    • A23K10/10Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A23K10/16Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
    • A23K10/18Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K20/00Accessory food factors for animal feeding-stuffs
    • A23K20/10Organic substances
    • A23K20/105Aliphatic or alicyclic compounds
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K20/00Accessory food factors for animal feeding-stuffs
    • A23K20/10Organic substances
    • A23K20/111Aromatic compounds
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K50/00Feeding-stuffs specially adapted for particular animals
    • A23K50/80Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66Microorganisms or materials therefrom
    • A61K35/74Bacteria
    • A61K35/741Probiotics
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00Medicinal preparations containing antigens or antibodies
    • A61K39/02Bacterial antigens
    • A61K39/104Pseudomonadales, e.g. Pseudomonas
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00Medicinal preparations containing antigens or antibodies
    • A61K39/02Bacterial antigens
    • A61K39/107Vibrio
    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02ATECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A40/00Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
    • Y02A40/80Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
    • Y02A40/81Aquaculture, e.g. of fish

Description

METHOD FOR TREATING AND/OR PREVENTING A SEA LICE INFECTION IN A TELEOST
TECHNICAL FIELD
The present document is directed to combating sea lice infestations on teleosts. More particularly, the present document discloses a probiotic composition which prevents and treats sea lice infestations in teleosts.
BACKGROUND OF THE INVENTION
Fish farming involves raising fish in tanks or enclosures. In aquaculture, freshwater, saltwater or anadromic populations, such as fish, crustaceans and shellfish, are raised under controlled conditions. Mariculture is a sub-branch of aquaculture where marine organisms are cultivated in the open ocean or an enclosed section of the ocean, or alternatively in ponds, tanks and the like filled with seawater.
Teleosts are the largest infraclass of the ray-finned fishes. There are over 26000 species, and they separated from other fish, such as sturgeons and bowfins, around 300 million years ago. The teleosts thus share many biological traits, including skin and mucus. Research indicates that the mucus cells are based on primordially conserved principles, that could be common to mammals and fish alike. Within the teleosts and salmonids the skin epidermis should thus have similar properties.
The growth and health of fish raised by intensive aquaculture is dependent on that sufficient oxygen and clean water with optimal levels of carbon dioxide, ammonia and with feasible pH can be provided. Also, a sufficient amount of feed with a high content of protein and a well-balanced supply of amino acids is crucial to obtain a commercially durable result. Attempts to increase the growth of farmed fish have generally focused on changing the contents of the feed used.
Farmed fish are often contained at high population densities which increase the risk for infections by e.g. parasites such as fish lice, intestinal worms, fungi, virus and bacteria.
Sea lice belong to the family of copepods (small crustaceans) within the order Siphonostomatoida, family Caligidae. The two main genera are the Lepeophtheirus and Caligus genera which infect marine fish. Many sea louse species are specific with regard to host genera. For example, L. salmonis has high specificity for salmonids, including farmed Atlantic salmon (Salmo salar) but it can also parasitize on other salmonids, including brown trout (sea trout: Salmo trutta), Arctic char (Salvelinus alpinus), and all species of Pacific salmon.
Sea lice feed on the fish and large numbers of lice on the same fish and, or just a couple of lice on a juvenile fish, can be harmful or fatal. Sea lice feed on the mucus, epidermal tissue, and blood of host marine fish and can cause fin damage, skin erosion, constant bleeding, and open wounds creating a pathway for other pathogens. More damage may be caused when infected fish jump or scrape along nets in an attempt to dislodge any irritating lice. In addition to the suffering from the fish, wholesalers and consumers prefer to buy fish that are free from sea lice, as infested fish are not aesthetically pleasing. This is of course a great problem in fish farming, but the sea lice may also spread from fish farms and infect wild fish.
Sexually mature and fertilized female lice release several hundred eggs by hatching. Free-living stages are referred to as nauplii (2 stages) and the copepodids along with chalimus (2 stages) are stationary/attached. All stages, including the movable stages (preand adult lice) feed on skin, mucus, blood on/from the fish and the fish’s microbiota.
Treatment and prevention of sea lice infestations generally involve the use of different chemical agents, such as organophosphates, carbamates, pyrethroids, pyrethrins, synergists, insect growth regulating chemicals or avermectins. However, such use is undesirable, e.g. due to the spread of these toxic agents to the environment.
Development of drug resistance in the sea lice has terminated the use of chemical delousing in several salmon farming areas and other delousing means as use of freshwater and mechanical delousing has increased in recent years to replace the use of chemical delousing. Increased frequencies of delousing in particular with mechanical methods during the warmer season results in more stress and more disease and mortality among the farmed salmon.
WO2018007632 and WO2019135009 disclose methods for increasing the growth rate and weight of fish or treating and preventing microbial infections respectively by administering probiotic bacteria such as Aliivibrio njordis, Aliivibrio balderis, and/or Aliivibrio nannie to the fish, such as by topical administration. There is however, no evidence that probiotics have any effect against ectoparasitic infestation. WO2015155293 discloses a feed composition comprising saponins and nucleotides, prebiotics and unnamed probiotics for use in the prevention and treatment of an ectoparasitic infection or infestations in fish.
To protect the wild salmonid species in the salmon farming regions there has been made strict regulations from the governmental authorities to monitor sea lice infestation weekly and treat sea lice infestation. In Norway farmed salmonids is mandatorily deloused when the level of sea lice passes 0.1 sea lice per salmon individual, a level of infestation that is not biologically relevant on a mature individual salmon. These regulations results in frequent sea lice treatments which stresses the farmed salmon.
Cleaner fish that are provided into the aquaculture pens to eat the sea lice from the skin of the primary farmed fish as salmonids are commonly used. The cleaner fish may be teleosts such as wild caught wrasse species as Goldsinny-wrasse (Ctenolabris rupestris), Ballan wrasse (Labrus bergylta) and corkwing wrasse (Symphodus melops) or lump fish (Cyclopterus lumpus). Recently farming of lump fish for use as cleaner fish has been replacing the wild catch and farming of Ballan wrasse has also been developed. There is an animal welfare issue linked to the use of cleaner fish since most of the cleaner fish dies in the pens before the salmon is reaching slaughter size. Little is known about the reasons for the loss of cleaner fish but a large factor is the development of infectious diseases including ulcers, fin rot, metazoan and protistan parasites.
The use of cleaner fish reduces the need of chemical or mechanical delousing episodes efficiently, but not fully and if delousing occurs by chemical, mechanical or thermal delousing the cleaner fish may die.
The economical impact of the sea lice infestations and the need of delousing has doubled the production costs from 1.5 to 3 euro per kilo Atlantic salmon produced the last 20 years in Norway.
An object of the present invention is thus to overcome or at least mitigate some of the problems described above.
SUMMARY
The present document is directed to a probiotic composition comprising probiotic bacteria of the species Aliivibrio njordis and/or Aliivibrio balderis, and optionally Aliivibrio nannie, for use in the treatment and/or prevention of a sea lice infestation in a teleost.
The probiotic composition may further comprise probiotic bacteria of the species Psychrobacter piscimesodermis, Psychrobacter piscimesenchymalis, Psychrobacter piscisubcutanea, Pseudomonas salmosubcutaneae, Pseudomonas salmosubpectoralis and/or Pseudomonas salmointermuscularis.
The present document also discloses a probiotic composition comprising probiotic bacteria of the species Aliivibrio njordis and/or Aliivibrio balderis, and optionally Aliivibrio nannie for use in the treatment and/or prevention of a sea lice infestation in a teleost, wherein said probiotic composition is administered to said teleost before, during or after administration of one or more further probiotic composition(s) comprising one or more of a probiotic bacterium selected from the group consisting of bacteria of the species Psychrobacter piscimesodermis, Psychrobacter piscimesenchymalis, Psychrobacter piscisubcutanea, Pseudomonas salmosubcutaneae, Pseudomonas salmosubpectoralis and/or Pseudomonas salmointermuscularis.
The probiotic composition may comprise probiotic bacteria of two or more different strains of the same bacterial species.
The teleost may be a marine teleost or a fresh water teleost.
The teleost may be of the family Salmonidae, such as salmon, trout, and chars. The teleost of the family Salmonidae may e.g. be Atlantic salmon (Salmo salar), brown trout (sea trout: Salmo trutta), Arctic char (Salvelinus alpinus), and/or any species of Pacific salmon, such as (Coho (Silver) salmon (Oncorhynchus kisutch), Chinook (King) (Oncorhynchus tshawytscha), pink (Humpy) salmon (Oncorhynchus gorbuscha), chum (Dog) salmon (Oncorhynchus keta), sockeye (Oncorhynchus nerka), cutthroat (Oncorhynchus clarki clarki) and steelhead (rainbow trout) (Oncorhynchus mykiss).
The teleost may also be Atlantic cod, cleaner fish like lump fish and wrasses and freshwater fish as carps and perch.
The sea lice may be of the species Lepeophtheirus salmonis, Caligus rogercresseyi and/or Caligus elongatus.
The probiotic composition may be administered in combination with a chemical parasitic infection treatment agent, such as an organophosphate, a carbamate, a pyrethroid, a pyrethrin, a synergist, an insect growth regulating chemical and/or an avermectin.
The probiotic composition may be administered via bath or dip administration in salt, fresh or brackish water, by oral administration, or injection, such as bath administration for a time period of 1 second to 5 hours, such as 1 second to 2 hours, such as 1 seconds to 1 hour, such as 30 seconds to 1 hour or 1 minute to 30 minutes.
The probiotic composition may be administered at least 10 days, such as at least 15 days, 20 days, 25 days, 30 days, or 40 days, before exposure of the teleost to the sea lice.
Other features and advantages of the invention will be apparent from the following detailed description, drawings, examples, and from the claims.
DEFINITIONS
Sea lice attaching infesting teleosts comprise:
● Family Caligidae with many different genera (37) with close to 600 species. Genus Lepeophtheirus with 162 species with L. salmonis as the most important for salmonids. Genus Caligus with 268 species with C. elongatus Norway (mostly Northern Norway) and Northern Atlantic and C. rogercresseyi (Pacific coasts including Chile and British Columbia).
Examples of Salmonides in which a sea lice infection may be treated and/or prevented with a probiotic composition of the present document comprise the species (reproduced from FishBase under CC 3.0 license from
https://www.fishbase.se/identification/specieslist.php?famcode=76):
Brachymystax lenok, Brachymystax savinovi, Brachymystax tumensis, Coregonus albellus, Coregonus albula, Coregonus alpenae, Coregonus alpinus, Coregonus anaulorum, Coregonus arenicolus, Coregonus artedi, Coregonus atterensis, Coregonus austriacus, Coregonus autumnalis, Coregonus baerii, Coregonus baicalensis, Coregonus baunti, Coregonus bavaricus, Coregonus bezola, Coregonus candidus, Coregonus chadary, Coregonus clupeaformis, Coregonus clupeoides, Coregonus confusus, Coregonus danneri, Coregonus duplex, Coregonus fatioi, Coregonus fera, Coregonus fontanae, Coregonus gutturosus, Coregonus heglingus, Coregonus hiemalis, Coregonus hoferi, Coregonus holsata, Coregonus hoyi, Coregonus huntsmani, Coregonus johannae, Coregonus kiletz, Coregonus kiyi, Coregonus ladogae, Coregonus laurettae, Coregonus lavaretus, Coregonus lucinensis, Coregonus lutokka, Coregonus macrophthalmus, Coregonus maraena, Coregonus maraenoides, Coregonus maxillaris, Coregonus megalops, Coregonus migratorius, Coregonus muksun, Coregonus nasus, Coregonus nelsonii, Coregonus nigripinnis, Coregonus nilssoni, Coregonus nipigon, Coregonus nobilis, Coregonus oxyrinchus, Coregonus palaea, Coregonus pallasii, Coregonus peled, Coregonus pennantii, Coregonus pidschian, Coregonus pollan, Coregonus pravdinellus, Coregonus reighardi, Coregonus renke, Coregonus restrictus, Coregonus sardinella, Coregonus stigmaticus, Coregonus subautumnalis, Coregonus suidteri, Coregonus trybomi, Coregonus tugun, Coregonus ussuriensis, Coregonus vandesius, Coregonus vessicus, Coregonus wartmanni, Coregonus widegreni, Coregonus zenithicus, Coregonus zuerichensis, Coregonus zugensis, Hucho bleekeri, Hucho hucho, Hucho ishikawae, Hucho taimen, Oncorhynchus aguabonita, Oncorhynchus apache, Oncorhynchus chrysogaster, Oncorhynchus clarkii, Oncorhynchus formosanus, Oncorhynchus gilae, Oncorhynchus gorbuscha, Oncorhynchus iwame, Oncorhynchus kawamurae, Oncorhynchus keta, Oncorhynchus kisutch, Oncorhynchus masou, Oncorhynchus mykiss, Oncorhynchus nerka, Oncorhynchus rhodurus, Oncorhynchus tshawytscha, Parahucho perryi, Prosopium abyssicola, Prosopium coulterii, Prosopium cylindraceum, Prosopium gemmifer, Prosopium spilonotus, Prosopium williamsoni, Salmo abanticus, Salmo akairos, Salmo aphelios, Salmo balcanicus, Salmo carpio, Salmo caspius, Salmo cenerinus, Salmo cettii, Salmo chilo, Salmo ciscaucasicus, Salmo coruhensis, Salmo dentex, Salmo euphrataeus, Salmo ezenami, Salmo farioides, Salmo ferox, Salmo fibreni, Salmo ischchan, Salmo kottelati, Salmo labecula, Salmo labrax, Salmo letnica, Salmo lourosensis, Salmo lumi, Salmo macedonicus, Salmo macrostigma, Salmo marmoratus, Salmo montenigrinus, Salmo multipunctata, Salmo nigripinnis, Salmo obtusirostris, Salmo ohridanus, Salmo okumusi, Salmo opimus, Salmo pallaryi, Salmo pelagonicus, Salmo pellegrini, Salmo peristericus, Salmo platycephalus, Salmo rhodanensis, Salmo rizeensis, Salmo salar, Salmo schiefermuelleri, Salmo stomachicus, Salmo taleri, Salmo tigridis, Salmo trutta, Salmo viridis, Salmo visovacensis, Salmo zrmanjaensis, Salvelinus agassizii, Salvelinus albus, Salvelinus alpinus, Salvelinus anaktuvukensis, Salvelinus andriashevi, Salvelinus boganidae, Salvelinus colii, Salvelinus confluentus, Salvelinus curilus, Salvelinus czerskii, Salvelinus drjagini, Salvelinus elgyticus, Salvelinus evasus, Salvelinus faroensis, Salvelinus fimbriatus, Salvelinus fontinalis, Salvelinus gracillimus, Salvelinus grayi, Salvelinus gritzenkoi, Salvelinus inframundus, Salvelinus jacuticus, Salvelinus japonicus, Salvelinus killinensis, Salvelinus krogiusae, Salvelinus kronocius, Salvelinus kuznetzovi, Salvelinus lepechini, Salvelinus leucomaenis, Salvelinus levanidovi, Salvelinus lonsdalii, Salvelinus mallochi, Salvelinus malma, Salvelinus maxillaris, Salvelinus murta, Salvelinus namaycush, Salvelinus neiva, Salvelinus neocomensis, Salvelinus obtusus, Salvelinus perisii, Salvelinus profundus, Salvelinus salvelinoinsularis, Salvelinus schmidti, Salvelinus struanensis, Salvelinus taimyricus, Salvelinus taranetzi, Salvelinus thingvallensis, Salvelinus tolmachoffi, Salvelinus umbla, Salvelinus vasiljevae, Salvelinus willoughbii, Salvelinus youngeri, Salvethymus svetovidovi, Stenodus leucichthys, Stenodus nelma, Thymallus arcticus, Thymallus baicalensis, Thymallus brevipinnis, Thymallus brevirostris, Thymallus burejensis, Thymallus flavomaculatus, Thymallus grubii, Thymallus mertensii, Thymallus nigrescens, Thymallus pallasii, Thymallus svetovidovi, Thymallus thymallus, Thymallus tugarinae and Thymallus yaluensis.
Examples of sea lice, the infestation of which may be treated and/or prevented by administration of the probiotic composition disclosed herein comprise Lepeophtheirus acutus, Lepeophtheirus aesopus, Lepeophtheirus alvaroi, Lepeophtheirus anguilli, Lepeophtheirus appendiculatus, Lepeophtheirus argentus, Lepeophtheirus atypicus, Lepeophtheirus azoricus, Lepeophtheirus bagri, Lepeophtheirus bifidus, Lepeophtheirus bifurcatus, Lepeophtheirus bonaci, Lepeophtheirus brachyurus, Lepeophtheirus breviventris, Lepeophtheirus bychowskyi, Lepeophtheirus chaenichthyis, Lepeophtheirus chantoni, Lepeophtheirus chilensis, Lepeophtheirus clarionensis, Lepeophtheirus confusum, Lepeophtheirus constrictus, Lepeophtheirus cossyphi, Lepeophtheirus crabro, Lepeophtheirus crassus, Lepeophtheirus cuneifer, Lepeophtheirus curtus, Lepeophtheirus dissimulatus, Lepeophtheirus distinctus, Lepeophtheirus edwardsi, Lepeophtheirus elegans, Lepeophtheirus eminens, Lepeophtheirus epinepheli, Lepeophtheirus erecsoni, Lepeophtheirus etelisi, Lepeophtheirus europaensis, Lepeophtheirus exilipes, Lepeophtheirus exsculptus, Lepeophtheirus formosanus, Lepeophtheirus frecuens, Lepeophtheirus furcatus, Lepeophtheirus goniistii, Lepeophtheirus grohmanni, Lepeophtheirus gusevi, Lepeophtheirus hapalogenyos, Lepeophtheirus hastatus, Lepeophtheirus heegaardi, Lepeophtheirus hexagrammi, Lepeophtheirus hidekoi, Lepeophtheirus hippoglossi, Lepeophtheirus histiopteridi, Lepeophtheirus hospitalis, Lepeophtheirus hummi, Lepeophtheirus intercurreus, Lepeophtheirus interitus, Lepeophtheirus kabatai, Lepeophtheirus krishnai, Lepeophtheirus lagocephali, Lepeophtheirus lalandei, Lepeophtheirus lateolabraxi, Lepeophtheirus latigenitalis, Lepeophtheirus lewisi, Lepeophtheirus lichiae, Lepeophtheirus litus, Lepeophtheirus longiabdominis, Lepeophtheirus longicaudus, Lepeophtheirus longipalpus, Lepeophtheirus longipes, Lepeophtheirus longispinosus, Lepeophtheirus longiventralis, Lepeophtheirus marcepes, Lepeophtheirus marginatus, Lepeophtheirus molvae, Lepeophtheirus monacanthus, Lepeophtheirus mugiloidis, Lepeophtheirus muraenae, Lepeophtheirus nanaimoensis, Lepeophtheirus natalensis, Lepeophtheirus nordmanni, Lepeophtheirus oblitus, Lepeophtheirus palliatus, Lepeophtheirus paralichthydis, Lepeophtheirus parvicruris, Lepeophtheirus parviventris, Lepeophtheirus parvulus, Lepeophtheirus parvus, Lepeophtheirus paulus, Lepeophtheirus pectoralis, Lepeophtheirus perpes, Lepeophtheirus platensis, Lepeophtheirus plectropomi, Lepeophtheirus plotosi, Lepeophtheirus pollachius, Lepeophtheirus polyprioni, Lepeophtheirus pravipes, Lepeophtheirus quadratus, Lepeophtheirus remiopsis, Lepeophtheirus renalis, Lepeophtheirus rhinobati, Lepeophtheirus robertae, Lepeophtheirus robustus, Lepeophtheirus rotundatus, Lepeophtheirus rotundipes, Lepeophtheirus rotundiventris, Lepeophtheirus schaadti, Lepeophtheirus scutiger, Lepeophtheirus sekii, Lepeophtheirus selkirki, Lepeophtheirus semicossyphi, Lepeophtheirus sheni, Lepeophtheirus shiinoi, Lepeophtheirus sigani, Lepeophtheirus simplex, Lepeophtheirus spatha, Lepeophtheirus spinifer, Lepeophtheirus sturionis, Lepeophtheirus suhmi, Lepeophtheirus tamladus, Lepeophtheirus tenuis, Lepeophtheirus thompsoni, Lepeophtheirus tuberculatus, Lepeophtheirus uluus, Lepeophtheirus unispinosus, Lepeophtheirus yanezi, Lepeophtheirus zbigniewi, Caligus abigailae, Caligus absens, Caligus acanthopagri, Caligus adanensis, Caligus aduncus, Caligus affinis, Caligus afurcatus, Caligus alaihi, Caligus alepicolus, Caligus amblygenitalis, Caligus antennatus, Caligus apodus, Caligus arii, Caligus ariicolus, Caligus asperimanus, Caligus asymmetricus, Caligus atromaculatus, Caligus balistae, Caligus belones, Caligus berychis, Caligus biaculeatus, Caligus bicycletus, Caligus bifurcus, Caligus biseriodentatus, Caligus bocki, Caligus bonito, Caligus brevicaudatus, Caligus brevicaudus, Caligus brevipedis, Caligus brevis, Caligus buechlerae, Caligus callaoensis, Caligus callyodoni, Caligus calotomi, Caligus carangis, Caligus centrodonti, Caligus chamelensis, Caligus cheilodactyli, Caligus chelifer, Caligus chiastos, Caligus chorinemi, Caligus chrysophrysi, Caligus clavatus, Caligus clemensi, Caligus confusus, Caligus constrictus, Caligus cookeoli, Caligus cordiventris, Caligus cordyla, Caligus cornutus, Caligus coryphaenae, Caligus costatus, Caligus cresseyorum, Caligus crusmae, Caligus curtus, Caligus cybii, Caligus dactylopteni, Caligus dakari, Caligus dampieri, Caligus dasyaticus, Caligus debueni, Caligus deformis, Caligus diaphanus, Caligus dicentrarchi, Caligus dieuzeidei, Caligus digitatus, Caligus dubius, Caligus elasmobranchi, Caligus eleutheronemi, Caligus elongatus, Caligus engraulidis, Caligus enormis, Caligus epidemicus, Caligus epinepheli, Caligus equulae, Caligus evelynae, Caligus eventilis, Caligus fajerae, Caligus fistulariae, Caligus flexispina, Caligus fortis, Caligus fronsuganinus, Caligus fugu, Caligus furcisetifer, Caligus glacialis, Caligus glandifer, Caligus grandiabdominalis, Caligus guerini, Caligus gurnardi, Caligus haemulonis, Caligus hamatus, Caligus hamruri, Caligus hemiconiati, Caligus hobsoni, Caligus hoplognathi, Caligus hottentotus, Caligus hyalinae, Caligus hyalinus, Caligus hyporhamphi, Caligus ignotus, Caligus ilhoikimi, Caligus inanis, Caligus infestans, Caligus inopinatus, Caligus irritans, Caligus isonyx, Caligus itacurussensis, Caligus jawahari, Caligus kabatae, Caligus kahawai, Caligus kala, Caligus kalumai, Caligus kanagurta, Caligus kapuhili, Caligus keralensis, Caligus kirti, Caligus klawei, Caligus kurochkini, Caligus kuwaitensis, Caligus labracis, Caligus lacustris, Caligus lagocephali, Caligus lalandei, Caligus laminatus, Caligus laticaudus, Caligus latigenitalis, Caligus latus, Caligus lessonius, Caligus lethrinicola, Caligus lichiae, Caligus ligatus, Caligus ligusticus, Caligus lini, Caligus littoralis, Caligus lobodes, Caligus lolligunculae, Caligus longiabdominis, Caligus longicaudatus, Caligus longicaudus, Caligus longipedis, Caligus longipes, Caligus longiramus, Caligus longirostris, Caligus longispinosus, Caligus lunatus, Caligus lutjani, Caligus macarovi, Caligus macoloricola, Caligus macrurus, Caligus malabaricus, Caligus mebachii, Caligus minimus, Caligus mordax, Caligus mortis, Caligus mugilis, Caligus mulli, Caligus musaicus, Caligus mutabilis, Caligus nanhaiensis, Caligus nataliae, Caligus nengai, Caligus neoaricolus, Caligus nibeae, Caligus nolani, Caligus novocaledonicus, Caligus nuenonnae, Caligus oculicola, Caligus ocyurus, Caligus ogawai, Caligus olsoni, Caligus omissus, Caligus orientalis, Caligus oviceps, Caligus pagelli, Caligus pageti, Caligus pagri, Caligus pagrosomi, Caligus pampi, Caligus paranengai, Caligus parapetalopsis, Caligus parvilatus, Caligus patulus, Caligus pauliani, Caligus pectinatus, Caligus pelagicus, Caligus pelamydis, Caligus penrithi, Caligus pharaonis, Caligus phipsoni, Caligus placidus, Caligus planktonis, Caligus platurus, Caligus platytarsis, Caligus polycanthi, Caligus pomacentrus, Caligus pomadasi, Caligus praecinctorius, Caligus praetextus, Caligus priacanthi, Caligus productus, Caligus pseudokalumai, Caligus pseudoproductus, Caligus pseudorhombi, Caligus pterois, Caligus punctatus, Caligus quadratus, Caligus quadrigenitalis, Caligus randalli, Caligus raniceps, Caligus rapax, Caligus regalis, Caligus reniformis, Caligus robustus, Caligus rogercresseyi, Caligus rotundigenitalis, Caligus rufimaculatus, Caligus saucius, Caligus savala, Caligus schistonyx, Caligus schlegeli, Caligus sciaenops, Caligus sclerotinosus, Caligus scribae, Caligus sensorius, Caligus sepetibensis, Caligus seriolae, Caligus seriolicolus, Caligus serratus, Caligus sibogae, Caligus sicarius, Caligus similis, Caligus solea, Caligus spinosus, Caligus stocki, Caligus stokesi, Caligus stromatei, Caligus subparvus, Caligus suffuscus, Caligus tanago, Caligus temnodontis, Caligus tenax, Caligus tenuicauda, Caligus tenuifurcatus, Caligus tenuis, Caligus tenuis, Caligus teres, Caligus tetrodontis, Caligus thyrsitae, Caligus torpedinis, Caligus trachynoti, Caligus triabdominalis, Caligus triangularis, Caligus tripedalis, Caligus truttae, Caligus turbidus, Caligus tylosuri, Caligus undulatus, Caligus uniartus, Caligus upenei, Caligus ventrosetosus, Caligus vexator, Caligus willungae, Caligus wilsoni, Caligus xystercus, Caligus zei, Caligus zylanica.
A “culture” includes all forms of bacterial culture, both in broth, on agar and in any other media. A “single culture” refers to a culture containing only one bacterial strain, i.e. a pure culture. A “mixed culture” refers to a culture wherein two or more bacterial strains, species and/or general are grown together or wherein two or more bacterial strains, species and/or genera are grown separately and thereafter mixed.
“CFU” stands for “colony forming units” which is a unit used for estimating the number of viable bacterial cells in a sample.
Milliliter is herein abbreviated with “ml” or “mL”.
BRIEF DESCRIPTION OF DRAWINGS
Figure 1: Probiotic application treatment (“Stembiont”) followed by lice challenge according to Example 1.
Figure 2: Results of cohabitant fish exposed to a probiotic composition (“Stembiont”) and control fish according to Example 2.
Figure 3: Results of non-cohabitant exposed to a probiotic composition (“Stembiont”) and control fish according to Example 2.
Figure 4: Goblet cells in four experiments. The treated fish have fewer cells with more mucus.
Figure 5: Deposit receipt and viability statement for Aliivibrio njordis (B1-25, 18-1/2013 mandib V11).
Figure 6: Deposit receipt and viability statement for Aliivibrio balderis (B1-24, 18-1/2013 kidn V12).
Figure 7: Deposit receipt and viability statement for Aliivibrio nannie (B8-24, 313/2013 kidn V13).
Figure 8: Deposit receipt and viability statement for Psychrobacter piscimesodermis Fisk 1, 41, 6/3-2014 Atl. Salm.
Figure 9: Deposit receipt and viability statement for Psychrobacter piscimesenchymalis Fisk 2, 42, 6/3-2014 Atl. Salm.
Figure 10: Deposit receipt and viability statement for Psychrobacter piscisubcutanea may for example be Psychrobacter piscisubcutanea Fisk 3, 43, 6/3-2014 Atl. Salm.
Figure 11: Deposit receipt and viability statement for Pseudomonas salmosubcutaneae Fisk 3, 13/5-2014, hb, Atl. Salm.
Figure 12: Deposit receipt and viability statement for Pseudomonas salmosubpectoralis Fisk 3, 13/5-2014, ba, Atl. salm.
Figure 13: Deposit receipt and viability statement for Pseudomonas salmointermuscularis Fisk 4, 13/5-2014, ha, Atl. salm
DETAILED DESCRIPTION
The present document is directed to treating and/or preventing sea lice infestations in teleosts. The present inventors surprisingly found that by administering probiotic bacteria to the teleosts, sea lice infestations could be prevented and/or treated.
The present document is thus directed to a probiotic composition comprising probiotic bacteria of one or both of the species Aliivibrio njordis and/or Aliivibrio balderis for use in the treatment and/or prevention of a sea lice infestation in a teleost. The present document is also directed to the use of probiotic bacteria of one or both of the species Aliivibrio njordis and Aliivibrio balderis for the preparation of a medicament in the form of a probiotic composition as disclosed for the treatment and/or prevention of a sea lice infestation in a teleost. The present document is further directed to a method for treating and/or preventing a sea lice infestation in a teleost, said method comprising administering a probiotic composition comprising probiotic bacteria of one or both of the species Aliivibrio njordis and Aliivibrio balderis to a teleost in need thereof. The probiotic composition, its preparation and administration will be further described in the below.
Preferably, the composition comprises both of the Aliivibrio njordis and Aliivibrio balderis species. A probiotic composition of the present document may also comprise one or more of a probiotic bacterium of the species Aliivibrio nannie, Psychrobacter piscimesodermis, Psychrobacter piscimesenchymalis, Psychrobacter piscisubcutanea, Pseudomonas salmosubcutaneae, Pseudomonas salmosubpectoralis and/or Pseudomonas salmointermuscularis. It is also possible to provide the probiotic bacteria in separate probiotic compositions which then are administered, simultaneously or sequentially, to a teleost. The same is true for the other probiotic bacterial species mentioned herein to be useful for the purpose of treating and/or preventing a sea lice infestation in a teleost, i.e. the probiotic bacteria can be provided in the same probiotic composition or in one or more separate probiotic compositions, each comprising one or more probiotic bacterial species and/or strains. Thus, unless otherwise stated, the term “probiotic composition for use in the treatment and/or prevention of a sea lice infestation”, and the like, refers to either a single probiotic composition comprising one or more probiotic bacterial species and/or strains or two or more probiotic compositions, each comprising one or more probiotic bacterial species and/or strains intended to be administered simultaneously or sequentially to a teleost. Likewise, the term “probiotic composition” as used herein refers to refers to either a single probiotic composition comprising one or more probiotic bacterial species and/or strains or two or more probiotic compositions, each comprising one or more probiotic bacterial species and/or strains intended to be administered simultaneously or sequentially to a teleost.
The probiotic bacteria of composition of the present document may consist of Aliivibrio njordis and/or Aliivibrio balderis and optionally one or more of a bacterial species selected from the group consisting Aliivibrio nannie, Psychrobacter piscimesodermis, Psychrobacter piscimesenchymalis, Psychrobacter piscisubcutanea, Pseudomonas salmosubcutaneae, Pseudomonas salmosubpectoralis and Pseudomonas
salmointermuscularis.
Thus, the present document also discloses a probiotic composition comprising probiotic bacteria of the species Aliivibrio njordis and/or Aliivibrio balderis, and optionally Aliivibrio nannie for use in the treatment and/or prevention of a sea lice infestation in a teleost, wherein said probiotic composition is administered to said teleost before, during or after administration of one or more further probiotic composition(s) comprising one or more of a probiotic bacterium selected from the group consisting of bacteria of the species Psychrobacter piscimesodermis, Psychrobacter piscimesenchymalis, Psychrobacter piscisubcutanea, Pseudomonas salmosubcutaneae, Pseudomonas salmosubpectoralis and/or Pseudomonas salmointermuscularis.
The present document also discloses the use of a a probiotic composition comprising probiotic bacteria of the species Aliivibrio njordis and/or Aliivibrio balderis, and optionally Aliivibrio nannie and a probiotic composition comprising one or more of a probiotic bacterium selected from the group consisting of bacteria of the species Psychrobacter piscimesodermis, Psychrobacter piscimesenchymalis, Psychrobacter piscisubcutanea, Pseudomonas salmosubcutaneae, Pseudomonas salmosubpectoralis and/or Pseudomonas salmointermuscularis for the preparation of a medicament in the form of a probiotic composition for the treatment and/or prevention of a sea lice infestation in a teleost, wherein said probiotic composition Aliivibrio njordis and/or Aliivibrio balderis, and optionally Aliivibrio nannie is administered before, during or after administration of the one or more further probiotic composition(s) comprising a probiotic bacterium selected from the group consisting of bacteria of the species Psychrobacter piscimesodermis, Psychrobacter piscimesenchymalis, Psychrobacter piscisubcutanea, Pseudomonas salmosubcutaneae, Pseudomonas salmosubpectoralis and/or Pseudomonas salmointermuscularis.
The present document further discloses a method for treating and/or preventing a sea lice infestation in a teleost wherein said method comprises administering a therapeutically effective amount of a probiotic composition comprising a probiotic bacterium of the species Aliivibrio njordis and/or Aliivibrio balderis, and optionally Aliivibrio nannie, before, during or after administration of a therapeutically effective amount of one or more further probiotic composition(s) comprising one or more of a probiotic bacterium selected from the group consisting of bacteria of the species Psychrobacter piscimesodermis, Psychrobacter piscimesenchymalis, Psychrobacter piscisubcutanea, Pseudomonas salmosubcutaneae, Pseudomonas salmosubpectoralis and/or Pseudomonas salmointermuscularis.
It is also possible to use two or more different strains of a probiotic bacterial species in the probiotic composition and/or probiotic bacteria of the same bacterial species/strain but grown under different growth conditions.
The probiotic composition is administered to teleosts in order to treat and/or prevent a sea lice infestation.
The teleost may be any kind of teleost, such as a teleost living in marine environments (salt or brackish water) or a teleost living in fresh water. Some teleosts, like teleosts of the family Salmonidae live in both fresh and salt water depending on where in their life cycle they are. Some teleosts, such as wrasse (Labridae) feed on sea lice of other teleosts.
Example of teleosts which can benefit from a treatment with the probiotic composition disclosed herein are teleosts of the family Salmonidae, such as salmon, trout, and chars. Examples of species of the family Salmonidae are Atlantic salmon (Salmo salar), brown trout (sea trout: Salmo trutta), Arctic char (Salvelinus alpinus), and/or any species of Pacific salmon, such as (Coho (Silver) salmon (Oncorhynchus kisutch), Chinook (King) (Oncorhynchus tshawytscha), pink (Humpy) salmon (Oncorhynchus gorbuscha), chum (Dog) salmon (Oncorhynchus keta), sockeye (Oncorhynchus nerka), cutthroat (Oncorhynchus clarki clarki) and steelhead (rainbow trout) (Oncorhynchus mykiss).
Further examples of marine teleosts are turbot, sea bass and sea bream. Other examples are Atlantic cod, cleaner fish like lump fish and wrasses and freshwater fish as carps and perch.
Without wishing to be bound by theory, one effect of the probiotic bacteria in the probiotic composition of the present document may be that they change the goblet cells of the teleosts so that these respond more efficiently, rapidly and/or with a higher response to an irritation from sea lice trying to attach to the skin or surface of the teleost with an increased flow of mucus, which prevents the sea lice from attaching.
The sea lice may e.g. be of the genera Lepeophtheirus salmonis, Caligus rogercresseyi and/or Caligus elongatus.
The probiotic bacteria may be administered separately or in any combination of two or more of the species and/or different strains of the different species. Bacteria of the different species and/or different strains may be used at any relative ratio. However, typically, the ratio is about 1:10 to about 10:1 between any two species/strains when the bacteria are used in combination. For example, the ratio between any two species/strains when the bacteria are used in combination may be 1:4 to 4:1, 1:3 to 3:1, 1:2 to 2:1 or 1:1. However, it may also be about 1:100 to 100:1.
Probiotic bacteria of the different species disclosed herein and/or different strains within the same species may be mixed before administration. The different species of probiotic bacteria and/or different strains within the same species may also be cultured together in the same culture. It is also possible to culture two or more strains of the same species together or in combination with one or more strains of another species.
Probiotic bacteria of the different species and/or different strains within the same species may be administered separately but simultaneously to the same population of teleosts. It is also possible to expose the teleost to probiotic bacteria of the different species disclosed herein and/or strains of one or more of the species sequentially by administering one or more probiotic bacterial species/strains before the addition of one or more further probiotic bacterial species/strains. If such a sequential administration of the probiotic bacteria is to be used, it is possible to add the bacteria sequentially but without removing previously added bacteria or to effect removal of previously added bacteria before new bacteria are added, e.g. by exchanging the volume of bacteria containing water for new water before addition of further bacteria.
Bacterial species and strains
The probiotic bacteria used in accordance with the present document are of the Aliivibrio njordis and/or Aliivibrio balderis species. Further, bacteria of the species Aliivibrio nannie may be used.
In addition to the above mentioned Aliivibrio species, probiotic bacteria of the species Psychrobacter piscimesodermis, Psychrobacter piscimesenchymalis, Psychrobacter piscisubcutanea, Pseudomonas salmosubcutaneae, Pseudomonas salmosubpectoralis and/or Pseudomonas salmointermuscularis may be used.
As mentioned above, the probiotic composition disclosed herein comprises probiotic bacteria of the species of the species Aliivibrio njordis and/or Aliivibrio balderis. Aliivibrio njordis may for example be Aliivibrio njordis strain B1-25, 18-1/2013 mandib V11, which has been deposited at National Collection of Industrial and Marine Bacteria and has been assigned accession number NCIMB 42593. Aliivibrio balderis may for example be Aliivibrio balderis B1-24, 18-1/2013 kidn V12, which has been deposited at National Collection of Industrial and Marine Bacteria and has been assigned accession number NCIMB 42592.
One exemplary strain of Aliivibrio nannie is Aliivibrio nannie B8-24, 313/2013 kidn V13, which has been deposited at National Collection of Industrial and Marine Bacteria and has been assigned accession number NCIMB 42594.
These exemplary strains of A. njordis, A. balderis and A. nannie, respectively, have been isolated in Norway and deposited according to the Budapest Treaty on June 17, 2016, at the National Collection of Industrial and Marine Bacteria (Ferguson Building, Craibstone Estate, Bucksburn, Aberdeen, AB219YA, Scotland, United Kingdom). Further details on the isolation and growth of these Aliivibrio strains are given in WO 2018/007632.
Psychrobacter piscimesodermis may for example be Psychrobacter piscimesodermis Fisk 1, 41, 6/3-2014 Atl. Salm, which has been deposited at National Collection of Industrial and Marine Bacteria and has been assigned accession number NCIMB 42947.
Psychrobacter piscimesenchymalis may for example be Psychrobacter piscimesenchymalis Fisk 2, 42, 6/3-2014 Atl. salm, which has been deposited at National Collection of Industrial and Marine Bacteria and has been assigned accession number NCIMB 42948. Psychrobacter piscisubcutanea may for example be Psychrobacter piscisubcutanea Fisk 3, 43, 6/3-2014 Atl. Salm, which has been deposited at National Collection of Industrial and Marine Bacteria and has been assigned accession number NCIMB 42949.
These exemplary species of Psychrobacter have been isolated in Norway and deposited according to the Budapest Treaty on January 4, 2018, at the National Collection of Industrial and Marine Bacteria (Ferguson Building, Craibstone Estate, Bucksburn, Aberdeen, AB219YA, Scotland, United Kingdom). Further details on the isolation and growth of these Psychrobacter strains are given in WO 2019/135009.
Pseudomonas salmosubcutaneae may for example be Pseudomonas salmosubcutaneae Fisk 3, 13/5-2014, hb, Atl. Salm., which has been deposited at National Collection of Industrial and Marine Bacteria and has been assigned accession number 43330.
Pseudomonas salmosubpectoralis may for example be Pseudomonas salmosubpectoralis Fisk 3, 13/5-2014, ba, Atl. salm., which has been deposited at National Collection of Industrial and Marine Bacteria and has been assigned accession number 43331.
Pseudomonas salmointermuscularis may for example be Pseudomonas salmointermuscularis Fisk 4, 13/5-2014, ha, Atl. salm., which has been deposited at National Collection of Industrial and Marine Bacteria and has been assigned accession number 43332.
These three exemplary species of Pseudomonas have been isolated in Norway and deposited according to the Budapest Treaty on December 20, 2018, at the National Collection of Industrial and Marine Bacteria (Ferguson Building, Craibstone Estate, Bucksburn, Aberdeen, AB21159YA, Scotland, United Kingdom). Further details on the isolation and growth of these Pseudomonas strains are given in WO 2019/135009.
Preparation of the probiotic bacterial composition
The probiotic composition comprising Aliivibrio njordis and/or Aliivibrio balderis and optionally additional bacterial species as described elsewhere herein may be prepared by culturing the different bacterial species and/or strains of the same bacterial species together. It is also possible to grow the different bacterial species and/or strains in separate cultures and mix them before administration to a teleost. Also, it is possible to grow the bacteria separately and administer them to a teleost without prior mixing, such by simultaneous or consecutive administration. Further, it is possible to grow two or more different bacterial species and/or strains of the same bacterial species together and mix with one or more culture(s) of one or more bacterial species and/or strains or to administer such cultures separately (simultaneously or consecutively) to a teleost.
The probiotic bacteria may be used together with their used growth medium, i.e. the probiotic composition comprises the used growth medium. One advantage with this is that beneficial substances that are produced by the probiotic bacteria will be present in the probiotic composition. Alternatively, the probiotic bacteria may be separated from the use growth medium (e.g. by centrifugation or filtration) and resuspended in another medium, such as new growth medium, a buffer (such as phosphate buffered saline, PBS) or a salt solution (such as a sodium salt solution). In this case, the probiotic bacteria may be washed one or more times with e.g. growth medium, a buffer, such as PBS or saline (such as a sodium salt solution).
The concentration of the probiotic bacteria in the probiotic composition may vary but is typically in the range of from 10<2 >to 10<14 >CFU, such as from 10<8 >to 10<13>, 10<9 >to 10<13>, 10<10 >to 10<12>, 10<2 >to 10<7>, or from 10<5 >to 10<7 >CFU.
The ratio of the different probiotic bacterial species in the probiotic composition may vary depending on e.g. whether the bacteria are grown together or separately and mixed before or upon administration. Typically Aliivibrio njordis and/or Aliivibrio balderis are present in a ratio of about 1:10 to 10:1, such as about a ratio of 3:10 to 10:3 or a ratio of about 1:1. When the probiotic composition further comprises bacteria of the species A. nannie, each bacterial species is typically constitutes about 33% of the number of probiotic bacteria in the composition.
In addition to the probiotic bacteria, the probiotic composition may comprise e.g. a pharmaceutically acceptable excipient and/or adjuvant.
It is also possible to add a chemical parasitic infestation treatment agent, such as an organophosphate, a carbamate, a pyrethroid, a pyrethrin, a synergist, an insect growth regulating chemical and/or an avermectin to the probiotic composition comprising probiotic bacteria described herein and/or to the further probiotic composition described herein. Thus, the probiotic composition described herein and/or the further probiotic composition described herein may comprise a chemical parasitic infestation treatment agent such as those mentioned above. Additionally or alternatively, it is possible to administer such a chemical parasitic infection treatment agent in conjunction with the administration of the probiotic composition, such as simultaneously or sequentially with the administration of the probiotic composition.
It will be appreciated that the probiotic composition described herein may be provided as a kit of parts. For instance, there is provided a kit of parts comprising:
(i) a probiotic composition as described herein, and
(ii) one or more further probiotic composition(s) as described herein, and/or
(iii) instructions for use.
It will be appreciated that the instructions for use described herein may describe that the probiotic composition described herein should be administered before, during and/or after administration of the one or more further probiotic composition(s) as described herein. The administration may intend administration to teleosts.
Administration of the probiotic bacteria to teleosts
The probiotic composition may e.g. administered to teleosts via bath or dip administration in salt, fresh or brackish water, by oral administration, by oral administration or by injection, such as by intraperitoneal administration, intramuscular administration, or subcutaneous administration.
When the probiotic composition is administered via bath or dip administration, the probiotic bacteria are cultured in a suitable manner and then added to water. The water to which the bacteria are added may be the water that the fish are already contained in or may be water in another tank, cage or the like to which the fish are transferred. The water is typically the same kind of water that the fish are contained in depending on their growth stage. For e.g. salmon at the post smolt stage, the water is typically natural sea water. However, it is also possible to transfer the fish to another kind of water during the treatment. For example, salmon at the pre smolt stage, which live in fresh water, may be transferred to salt water, such as natural seawater, during the exposure to the probiotic bacteria and then moved back to the fresh water. The water to which the bacteria are added typically has a salinity of about 0.5 to about 4 weight%, such as about 0.9 to about 4 weight%, such as about 2 to 4 weight%, although the water may also be fresh water which has a much lower salinity. It is thus possible to use already propagated live cells to bath fish at lower salt concentrations down to fresh water at typical shorter time intervals not killing the probiotic bacteria.
The teleosts are exposed to the bathing water containing the probiotic bacteria for a time sufficient for enough bacteria to be administered to the teleosts to obtain the desired effects. This time will depend on e.g. the concentration of probiotic bacteria in the probiotic composition used, the type and status of the teleosts that are to be exposed etc. Typically, an exposure time of a few seconds (dip administration) to a couple of hours may be used, such as from about 1 second to about 5 hours, such as from about 1 second to about 2 hours, such as from about 1 second to about 1 hour, such as from about 30 seconds to about 1 hour or from about 1 minute to about 30 minutes. Increasing the concentration of probiotic bacteria in the water will generally decrease the exposure time needed.
The teleosts may be exposed to the probiotic bacteria a single time or the exposure may be repeated one or more times with different time intervals. It may be beneficial to administer the probiotic composition to the teleosts at least 10 days, such as at least 15 days, 20 days, 25 days, 30 days, or 40 days, before exposure of the teleost to the sea lice.
The probiotic bacteria may be administered to teleosts in a therapeutically effective amount. It will be appreciated that this amount will depend on the administration route used.
Typically, when dip or bath administration is used, the concentration of the probiotic bacteria in the bath is from about 10<4 >to about 10<13 >CFU/ml. The concentration when using dip administration generally has to be higher than if bath administration is used, due to the shorter exposure time when dip administration is used. For dip administration, the concentration is typically from about 10<7 >to about 10<13 >CFU/ml, such as from about 10<7 >to about 10<12 >CFU/ml, such as from about 10<9 >to about 10<12 >CFU/ml.When applying a bath for a short single treatment interval, such as a bathing lasting for about 15 to 60 minutes, a concentration of from about10<5 >to about 10<7 >may be suitable. For bath administration, the concentration is typically from about 10<6 >to about 10<11 >CFU/ml. Bacterial cultures prepared in fermenters may have a concentration of ca 10<13 >CFU/ml. A dilution of such a fermented culture of ca 1:100 to 1:600 may be suitable for application by bathing and a dilution of ca 1:20 for application by dipping. Adding probiotic bacteria at repeated intervals at lower concentrations down to the natural level in seawater may be beneficial to the bathed teleosts. A continuous infusion of probiotic bacteria at lower levels of concentration down to one cell/ml water may be beneficial to the teleosts.
Similarly the uptake of probiotic bacteria can be facilitated by injection of the bacteria, such as via injecting them through the abdominal wall exposing the serous linings of the peritoneal cavity for the probiotic bacteria. The concentration of bacteria used for such administration is typically from about 1 x 10<5 >to about 1 x 10<7 >CFU/ml, such as about 1 x 10<6 >CFU/ml. Typically about 0.1 ml is administered per teleost giving a dose of from about 1 x 10<4 >to about 1 x 10<6 >CFU, such as about 1 x 10<5 >CFU.
An additional way of administering probiotic bacteria is through oral intubation. It is considered that the immune cells in the distal part of the intestine is high in numbers and are able to transport bacteria across the intestinal wall. The concentration of bacteria in the probiotic composition used for such administration is typically from about 1 x 10<6 >to about 1 x 10<8 >CFU/ml, such as about 1 x 10<7 >CFU/ml. Typically about 0.1-0.2 ml of the probiotic composition is administered. Similarly intubation of the teleost through the mouth into the stomach is tested as an effective way of administering probiotic bacteria.
Administering probiotic bacteria to spawned eggs is an important and effective way of administering probiotic bacteria. Uptake of bacteria into the eggs and adhesion to the egg shells are mechanisms that are important in the protection of the eggs and early fry. Egg yolk fry and start-feeding fry is a group of fish that also very effectively can be protected by exposing to probiotic bacteria by bathing or dipping.
It is also possible to administer the probiotic bacteria in freeze-dried form.
Administration of probiotic bacteria to RAS facilities
Recirculation of water in fish farming facilities (RAS facilities) has increased due to different pressing reasons. The consumption of intake water to a farming facility can be down to only 5 % of a flow-through facility. This makes it easier to construct larger and more effective facilities independent of limitations of large water supplies. The low level of intake water also reduces the risk of attracting diseases through the external water sources. On the other side there is a large risk that “house strains” of bacteria may establish in the RAS facility through the bio-filter microbiota or through the biofilms established in the tanks and pipe systems.
RAS-facilities make it possible to keep a higher temperature in the RAS water which increases the speed of the growth of salmonids.
During and after the smoltification process marine water is commonly used to some extent to warm up the water or to increase the growth towards the sea transfer. If the level of marine water can be reduced in RAS the risk of getting marine pathogens into the facility can be reduced. Typically intake of marine water is made from depths that are below the salmon sea lice zone of the marine ecosystem both in RAS plants and in flow-through plants using marine water in particular in the post smolt period to reduce the time of farming in the open net cages in the sea. Even full-scale RAS systems for farming of Atlantic salmon from egg to slaughter in RAS facilities are now constructed.
The microbiotas of the various RAS facilities are studied so far to a low extent but it seems that ulcer and fin rot and depressed growth can be substantial problems in some facilities even to an extent that the whole system is stopped, sanitized and restarted. Direct use of probiotic bacteria in RAS systems can be effective ways of securing and increasing growth and reducing disease and mortality. The bio-filter is a complex organism with dominating groups of bacteria degrading organic material from the fish coming from feces and uneaten feed. The sedimentation unit will only sediment the larger particles. In addition to the microbiota in the bio-filter degrading organic compounds and binding nutrients there is a specific microbiota related to the health and welfare of the fish. This part of the microbiota in the bio-filter is only a minor part of the total microbiota in the bio-filter and also in the water of the fish tanks. Some RAS-facilities disinfect the water coming from the bio-filter before it enters the fish tanks. This may be an important factor controlling the risk of getting infectious diseases in the populations of fish. On the other hand such disinfection may also reduce the level of naturally probiotic bacteria in the plant.
Depending on the design of the various RAS-facilities, either they use fresh or marine water, the level of the normal probiotic bacteria in the water of the facility can be low or high. To be able to secure a stable high growth with healthy fish in RAS-systems it will be optimal to add probiotic bacteria either for fresh, brackish or salt water. If fish are bathed or supplied with probiotic bacteria they will shed bacteria into the water and in that way seed the water for uptake in fish later when the water return to the fish tanks. This means that using probiotic bacteria directly to the fish will also impact the bio-filter unit and then benefit the fish after the microbial processing of the water in the bio-filter.
Application of probiotic culture may be performed any place in the cycle of RAS-facilities in addition to be directed immediately to the fish, i.e. in the fish tank water, in the bio-filter unit, after the bio-filter unit and if a disinfection step is used after the bio-filter processing of the water after the disinfection but before the water reaches the fish tanks.
In summary probiotic bacteria can be administered in a large number of different ways depending on the design and management including stage of production of each facility.
If bacteria of more than one species and/or strain is to be used, the bacteria may be grown separately (i.e. in single cultures) or in the same culture (i.e. in mixed cultures). If grown separately, bacteria of the different cultures may be mixed before addition to the water or the bacteria of the different cultures may be added separately. Probiotic bacteria obtained from different cultures (independently of whether single or mixed cultures) may also be added separately but to the same volume of water for the fish to be exposed to the different bacteria simultaneously.
Administration preferably to be performed before infection but may also be possible during ongoing infection.
The invention will be further described in the following examples, which do not limit the scope of the invention described in the claims
EXPERIMENTAL SECTION
In all examples below the Aliivibrio njordis bacteria were of the strain Aliivibrio njordis strain B1-25, 18-1/2013 mandib V11, the Aliivibrio balderis bacteria were of the strain Aliivibrio balderis B1-24, 18-1/2013 kidn V12, and the Aliivibrio nannie bacteria were of the strain Aliivibrio nannie B8-24, 313/2013 kidn V13.
Example 1
Alteration of skin and potential utility of microbiome intervention in sea lice infestation avoidance was demonstrated by using probiotic application as a skin enhancing treatment in a laboratory study. Salmon were immersed in a solution comprising probiotic bacteria (“Stembiont”) for 60 seconds 12 days prior to challenge with 30 copepodids/fish for 30 minutes with oxygenation. The probiotic composition used was a bacterial composition comprising a mixed culture of Aliivibrio balderis and Aliivibrio njordis grown together (total concentration about 1.9 x 10<8 >cfu per ml of which about 70 % was A. njordis and about 30 % was A. balderis) cultured in Luria-Bertani broth (LB, 1% Bacto-tryptone, 0.5% Bactoyeast extract and 2.5% NaCl). The probiotic composition was used at a 1/20 dilution, i.e. the concentration of bacteria in the probiotic bath was 1/20 of the concentration of the stock culture. The culture was made by mixing the two cultures of the two bacterial species after making pure cultures in LB from frozen stock cultures before mixing the cultures before the final up-scaling of the culture volumes. The cultures can also be grown separately and mixed at the time of use. The probiotic bacterial cultures were used directly without removing the broth medium. The groups exposed to the probiotic composition, SB1 and SB2, consisted of 21-22 fish, and the control groups (exposed to sea lice but unexposed to the probiotic composition, Ctrl1 and Ctrl 2) were similarly set up and challenged with sea lice concurrently with the exposed fish.
Results: Lice counting was carried out at 28 days post challenge with sea lice (2 tanks with application of the probiotic composition and 2 parallel controls without probiotic application; Fig.1). The results showed a significant reduction in attached lice and a lack of feeding in the attached lice in the probiotic application groups (SB-1 and SB-2).
Example 2
Salmon (mean weight 150 g) were immersed for 35 min in a solution comprising a probiotic composition comprising Aliivibrio balderis and Aliivibrio njordis grown together (“Stembiont”, 1.23 x 10<10 >cfu/ml composition was used at a 1/20 dilution when bathing the fish<) >together with an anesthetic bath February 19, 2019, 159 days (5.5 months) before introduction of sea lice as a challenge. A. njordis constituted about 70% and A. balderis about 30% of the bacteria in the probiotic composition. Groups treated with the probiotic composition were pit-tagged with 12 mm RFID, itag 162, pit tagging was done under Benzoak Vet. anesthesia. Pit tags were ISO 11784 and 11785 approved, and IEC 8-26/29 tested. Pit tags were used as recommended from dealer, BTS-ID. Pit tags were injected with a N125 needle. Fish size was > 45g and >15 cm and the salmon immersed in probiotic bacteria cohabitated with control groups not exposed to the probiotic composition (Table 1, Figure 2) or in individual treated and control tanks as tabled (Table 2, Figure 3). The study also compared treatment efficacy in a co-habitant housing with lice treatments for passive immunity and vaccination.
Fish (mean weight 150 g) were challenged with 30 copepodids/fish for 30 minutes with oxygenation on July 24.07.19 and assessed for lice counts on August 27th, 34 days after the lice were introduced.
Table 1: Lice counts per cohabitant tank and per group, relative percent protection (RPP) 5.5 months after exposure to the probiotic composition.
* RPP is “relative percent protection” calculated 1 minus as the mean lice count divided by the mean lice count on the reference control group x 100 (as percent).
Table 2: Lice counts treatment group (not cohabitant), relative percent protection (RPP) 5.5 months after exposure to the probiotic composition.
Table 2
Example 3
Salmon 34 g were PIT tagged (see Example 2) and introduced to 180 L tanks 11.06.19. A probiotic composition comprising Aliivibrio balderis and Aliivibrio njordis (Composition 1 in Table 3 below) or Aliivibrio balderis, Aliivibrio njordis, and Aliivibrio nannie (Composition 2 in Table 3 below) was used as preventive empowerment as below in Table 3 in individual 180 L tanks as a part of a larger population (in the same manner as in examples 1 and 2).
For this example, Aliivibrio balderis and Aliivibrio njordis were co-cultured resulting in Composition 1 having about 30% A. balderis and about 70% A. njordis, while Aliivibrio nannie was cultured separately. To produce Composition 2, the A. nannie culture was added to Composition 1 resulting in a ratio of about 20% A. balderis and 47% A. njordis and about 33% A. nannie.
Composition 1 had a total bacterial count of about 7.2 x 10<8 >CFU/ml, while the Composition 2 had a total bacterial count of about 8.8 x 10<8 >CFU/ml. A 1/20 times dilution of Composition 1 and 2, respectively, was used for bathing the fish. The fish were bathed in the probiotic composition for 35 min.
To compare the effect of microbial enhancement to microbial depletion by use of an antimicrobial compound, Halamid (Chloramine T), fish in that group were given a Halamid treatment starting 17.06.19 weekly for four weeks in freshwater. All groups were introduced to SW (salt water) 10.07.19 and full SW 17.07.19.
The lice challenge (15 lice per fish):
24.07.19 Lice added
15.08.19 Check to confirm lice attach
27.08.19 Lice count
Five groups with duplicate, a total of 10 tanks.15 fish from each tank were moved to a separate holding tank and added 15 lice per fish (number calculated by size).
Table 3: Results
Per 27.08.19
● In general, the number of lice are very low.
● That there is a difference in the skin structure of the fish treated with Composition 1 or 2 as compared to untreated fish
● The lice normally bind from the dorsal fin and backwards, but in this experiment they mostly bind on the head.
● Some lice with lighter colors that seem easier to remove were observed in the probiotic enhanced groups.
17.09.19 check for egg strings
52 of the 150 fish in total were examined. Two lice were found, both in the Chalamus stage. This indicates that the lice have not evolved to the mature adult stage where egg strings evolve. Rather, the lice have detached when entering mobile adult stage (if they even reached adult stage).
Example 4
In this example the number of goblet cells in salmon with and without treatment with a probiotic composition was studied.
The Atlantic salmon smolts were empowered with a probiotic bacterial composition comprising A. balderis and A. njordis in connection with the vaccination operation 8 weeks before the smolts were transferred to sea water (March 2018). The probiotic composition was added to the anaesthetic bath (30 mg Benzoak Vet. pr. liter) before the presmolts were vaccinated by an automated vaccine robot. The probiotic composition was added to the anaesthetic bath in a dilution of 1:20 and the presmolt stayed in the bath for 30 to 40 seconds before automatically transferred to the vaccination robot. The bacterial concentration was 1 x 10 log<7 >cfu/ml in the mixed probiotic culture before dilution and 1 x 10 log<6 >cfu/ml in the application water after dilution. The probiotic bacteria were grown together and the ratio between A. balderis and A. njordis in the resulting culture was about 70 % and 30% respectively.
The vaccinated presmolts were reared for 8 weeks before transferred to sea water. The experienced personell at the smolt plant commented that there was many years since the smolts they produced had such a large layer of mucus produced when fish are handled. These observations lead to further trials with sea lice prevention from probiotic empowerment.
All data passed the D'Agostino & Pearson normality test, Shapiro-Wilk normality test, Brown-Forsythe test and Bartlett's test. Tukey's multiple comparisons test showed significance (P<0.0008) from the A2 group. A3 is a group of triploid fish, A2 is diploid, both from Ranfjord. B1-untreated and B1-treated are similar fish from Grytågå. The cells were counted in skin samples from the head and body of seven fish for each batch. For each fish three polygonal areas between 0.2 mm<2 >and 0.3 mm<2 >of the epidermis (the outermost layer of the skin) were sampled, one from the head, two from different parts of the body. The specimens were coloured with Periodic Acid Shiff (PAS) for the goblet cells to be distinct in images. The digital images were analysed by a skilled investigator.
Figure 4 shows the number of goblet cells (mucus cells) per square mm in salmon, analysed in GraphPad Prism v 7.03. The average number of goblet cells per mm<2 >± SD is: A3-treated: 914.7±203.7, A2-untreated: 1262 ± 186.8, B1-untreated: 08.11.181004 ± 257, B1-treated: 861.9 ± 180.5.
The untreated fish have more, but smaller goblet cells. In the treated fish, there are fewer cells, but they are larger with more mucus. Without wishing to be bound by theory, the effect of this may be that the fish respond with an increased flow of mucus to an irritation from sea lice trying to attach to the skin or surface of the fish.
It is to be understood that while the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.
Unless expressly described to the contrary, each of the preferred features described herein can be used in combination with any and all of the other herein described preferred features.
REFERENCES WO 2019/135009
ITEMS
1. A probiotic composition comprising probiotic bacteria of the species Aliivibrio njordis and/or Aliivibrio balderis for use in the treatment and/or prevention of a sea lice infestation in a teleost.
2. The probiotic composition for use according to item 1, wherein said probiotic composition comprises probiotic bacteria of both Aliivibrio njordis and Aliivibrio balderis.
3. The probiotic composition for use according to item 1 or 2, wherein said Aliivibrio njordis is Aliivibrio njordis strain B1-25, 18-1/2013 mandib V11, which has been deposited at National Collection of Industrial and Marine Bacteria and has been assigned accession number NCIMB 42593.
4. The probiotic composition for use of any one of the preceding items, wherein said Aliivibrio balderis is Aliivibrio balderis B1-24, 18-1/2013 kidn V12, which has been deposited at National Collection of Industrial and Marine Bacteria and has been assigned accession number NCIMB 42592.
5. The probiotic composition for use according to any one of the preceding items, wherein said probiotic composition further comprises probiotic bacteria of the species Aliivibrio nannie, such as Aliivibrio nannie B8-24, 313/2013 kidn V13, which has been deposited at National Collection of Industrial and Marine Bacteria and has been assigned accession number NCIMB 42594.
6. The probiotic composition for use according to any one of the preceding items, wherein said probiotic composition further comprises probiotic bacteria of the species Psychrobacter piscimesodermis, Psychrobacter piscimesenchymalis, Psychrobacter piscisubcutanea, Pseudomonas salmosubcutaneae, Pseudomonas salmosubpectoralis and/or Pseudomonas salmointermuscularis.
7. The probiotic composition for use according to item 6, wherein said Psychrobacter piscimesodermis is Psychrobacter piscimesodermis Fisk 1, 41, 6/3-2014 Atl. Salm, which has been deposited at National Collection of Industrial and Marine Bacteria and has been assigned accession number NCIMB 42947.
8. The probiotic composition for use according to item 6 or 7, wherein said Psychrobacter piscimesenchymalis is Psychrobacter piscimesenchymalis Fisk 2, 42, 6/3-2014 Atl. salm, which has been deposited at National Collection of Industrial and Marine Bacteria and has been assigned accession number NCIMB 42948.
9. The probiotic composition for use according to any one of items 6-8, wherein said Psychrobacter piscisubcutanea is Psychrobacter piscisubcutanea Fisk 3, 43, 6/3-2014 Atl. Salm, which has been deposited at National Collection of Industrial and Marine Bacteria and has been assigned accession number NCIMB 42949.
10. The probiotic composition for use according to any one of items 6-9, wherein said Pseudomonas salmosubcutaneae is Pseudomonas salmosubcutaneae Fisk 3, 13/5-2014, hb, Atl. Salm, which has been deposited at National Collection of Industrial and Marine Bacteria and has been assigned accession number 43330.
11. The probiotic composition for use according to any one of items 6-10, wherein said Pseudomonas salmosubpectoralis is Pseudomonas salmosubpectoralis Fisk 3, 13/5-2014, ba, Atl. salm., which has been deposited at National Collection of Industrial and Marine Bacteria and has been assigned accession number 43331.
12. The probiotic composition for use according to any one of items 6-11, wherein said Pseudomonas salmointermuscularis is Pseudomonas salmointermuscularis Fisk 4, 13/5-2014, ha, Atl. salm., which has been deposited at National Collection of Industrial and Marine Bacteria and has been assigned accession number 43332.
13. The probiotic composition for use according to item 1 or 2, wherein said probiotic composition is administered to said teleost before, during or after administration of one or more further probiotic composition(s) comprising one or more of a probiotic bacterium as defined in any one of items 3-12.
14. The probiotic composition for use according to any one of the preceding items, wherein said probiotic composition comprises probiotic bacteria of two or more different strains of the same bacterial species.
15. The probiotic composition for use according to any one of the preceding items, wherein said teleost is a marine teleost.
16. The probiotic composition for use according to any one of items 1-14, wherein said teleost is a fresh water teleost.
17. The probiotic composition for use according to any one of the preceding items, wherein said teleost is of the family Salmonidae, such as salmon, trout, and chars.
18. The probiotic composition for use according to item 16, wherein said teleost of the family Salmonidae is Atlantic salmon (Salmo salar), brown trout (sea trout: Salmo trutta), Arctic char (Salvelinus alpinus), and/or any species of Pacific salmon, such as (Coho (Silver) salmon (Oncorhynchus kisutch), Chinook (King) (Oncorhynchus tshawytscha), pink (Humpy) salmon (Oncorhynchus gorbuscha), chum (Dog) salmon (Oncorhynchus keta), sockeye (Oncorhynchus nerka), cutthroat
(Oncorhynchus clarki clarki) and steelhead (rainbow trout) (Oncorhynchus mykiss).
19. The probiotic composition for use according to any one of items 1-16, wherein said teleost is Atlantic cod, cleaner fish like lump fish and wrasses and freshwater fish as carps and perch.
20. The probiotic composition for use according to any one of the preceding items, wherein said sea lice is of the species Lepeophtheirus salmonis, Caligus rogercresseyi and/or Caligus elongatus.
21. The probiotic composition for use according to any one of the preceding items, wherein said probiotic composition further comprises a pharmaceutically acceptable excipient and/or adjuvant.
22. The probiotic composition for use according to any one of the preceding items, wherein said probiotic composition is administered in combination with a chemical parasitic infection treatment agent, such as an organophosphate, a carbamate, a pyrethroid, a pyrethrin, a synergist, an insect growth regulating chemical and/or an avermectin.
23. The probiotic composition for use according to any one of the preceding items, wherein said probiotic composition is administered via bath or dip administration in salt, fresh or brackish water, by oral administration, by oral administration or by injection, such as by intraperitoneal administration, intramuscular administration, or subcutaneous administration.
24. The probiotic composition for use according to item 23, wherein said bath administration takes place for a time period of from 1 second to 5 hours, such as from 1 second to 2 hours, such as from 1 second to 1 hour, such as from 30 seconds to 1 hour or from 1 minute to 30 minutes.
25. The probiotic composition for use according to any one of the preceding items, wherein the total concentration of bacteria in the probiotic composition is from 10<2 >to 10<14>, such as from 10<8 >to 10<13>, from 10<9 >to 10<13>, from 10<10 >to 10<12>, from 10<2 >to 10<7>, or from 10<5 >to 10<7>.
26. The probiotic composition for use according to any one of the preceding items, wherein said probiotic composition is administered at least 10 days, such as at least 15 days, 20 days, 25 days, 30 days, or 40 days, before exposure of the teleost to the sea lice.
27. Use of probiotic bacteria of one or both of the species Aliivibrio njordis and Aliivibrio balderis for the preparation of a medicament in the form of a probiotic composition for the treatment and/or prevention of a sea lice infestation in a teleost.
28. The use according to item 27, wherein said probiotic composition comprises probiotic bacteria of both Aliivibrio njordis and Aliivibrio balderis.
29. The use according to item 27 or 28, wherein said Aliivibrio njordis is Aliivibrio njordis strain B1-25, 18-1/2013 mandib V11, which has been deposited at National Collection of Industrial and Marine Bacteria and has been assigned accession number NCIMB 42593
30. The use according to any one items 27-29, wherein said Aliivibrio balderis is Aliivibrio balderis B1-24, 18-1/2013 kidn V12, which has been deposited at National Collection of Industrial and Marine Bacteria and has been assigned accession number NCIMB 42592.
31. The use according to any one of items 27-30, wherein said probiotic composition further comprises probiotic bacteria of the species Aliivibrio nannie, such as Aliivibrio nannie B8-24, 313/2013 kidn V13, which has been deposited at National Collection of Industrial and Marine Bacteria and has been assigned accession number NCIMB 42594.
32. The use according to any one of items 27-31, wherein said medicament further comprises probiotic bacteria of the species Psychrobacter piscimesodermis, Psychrobacter piscimesenchymalis, Psychrobacter piscisubcutanea, Pseudomonas salmosubcutaneae, Pseudomonas salmosubpectoralis and/or Pseudomonas salmointermuscularis.
33. The use according to item 32, wherein said Psychrobacter piscimesodermis is Psychrobacter piscimesodermis Fisk 1, 41, 6/3-2014 Atl. Salm, which has been deposited at National Collection of Industrial and Marine Bacteria and has been assigned accession number NCIMB 42947.
34. The use according to item 32 or 33, wherein said Psychrobacter piscimesenchymalis is Psychrobacter piscimesenchymalis Fisk 2, 42, 6/3-2014 Atl. salm, which has been deposited at National Collection of Industrial and Marine Bacteria and has been assigned accession number NCIMB 42948.
35. The use according to any one of items 32-34, wherein said Psychrobacter piscisubcutanea is Psychrobacter piscisubcutanea Fisk 3, 43, 6/3-2014 Atl. Salm, which has been deposited at National Collection of Industrial and Marine Bacteria and has been assigned accession number NCIMB 42949.
36. The use according to any one of items 32-35, wherein said Pseudomonas salmosubcutaneae is Pseudomonas salmosubcutaneae Fisk 3, 13/5-2014, hb, Atl. Salm, which has been deposited at National Collection of Industrial and Marine Bacteria and has been assigned accession number 43330.
37. The use according to any one of items 32-36, wherein said Pseudomonas salmosubpectoralis is Pseudomonas salmosubpectoralis Fisk 3, 13/5-2014, ba, Atl. salm., which has been deposited at National Collection of Industrial and Marine Bacteria and has been assigned accession number 43331.
38. The use according to any one of items 32-37, wherein said Pseudomonas salmointermuscularis is Pseudomonas salmointermuscularis Fisk 4, 13/5-2014, ha, Atl. salm., which has been deposited at National Collection of Industrial and Marine Bacteria and has been assigned accession number 43332.
39. Use of a a probiotic composition as defined in item 1 or 2 and a probiotic composition comprising one or more of a probiotic bacterium as defined in any one of items 3-12 for the preparation of a medicament in the form of a probiotic composition for the treatment and/or prevention of sea lice infestation in a teleost, wherein said probiotic composition as defined in item 1 or 2 is administered to said teleost before, during or after administration of one or more further probiotic composition(s) comprising a probiotic bacterium as defined in any one of items 3-12.
40. The use according to any one of items 27-39, wherein said probiotic composition comprises probiotic bacteria of two or more different strains of the same bacterial species.
41. The use according to any one of items 27-40, wherein said teleost is a marine teleost.
42. The use according to any one of items 27-40, wherein said teleost is a fresh water teleost.
43. The use according to any one of items 27-42, wherein said teleost is of the family Salmonidae, such as salmon, trout, and chars.
44. The use according to item 43, wherein said teleost of the family Salmonidae is Atlantic salmon (Salmo salar), brown trout (sea trout: Salmo trutta), Arctic char (Salvelinus alpinus), and/or any species of Pacific salmon, such as (Coho (Silver) salmon (Oncorhynchus kisutch), Chinook (King) (Oncorhynchus tshawytscha), pink (Humpy) salmon (Oncorhynchus gorbuscha), chum (Dog) salmon
(Oncorhynchus keta), sockeye (Oncorhynchus nerka), cutthroat (Oncorhynchus clarki clarki) and steelhead (rainbow trout) (Oncorhynchus mykiss).
45. The use according to any one of items 27-42, wherein said teleost is Atlantic cod, cleaner fish such as lump fish and wrasses and freshwater fish as carps and perch.
46. The use according to any one of items 27-45, wherein the sea lice is of the species Lepeophtheirus salmonis, Caligus rogercresseyi and/or Caligus elongatus.
47. The use according to any one of items 27-46, wherein said probiotic composition further comprises a pharmaceutically acceptable excipient and/or adjuvant.
48. The use according to any one of items 27-47, wherein said probiotic composition is administered in combination with a chemical parasitic infection treatment agent, such as an organophosphate, a carbamate, a pyrethroid, a pyrethrin, a synergist, an insect growth regulating chemical and/or an avermectin.
49. The use according to any one of items 27-48, wherein said probiotic composition is administered via bath or dip administration in salt, fresh or brackish water, by oral administration, by oral administration or by injection, such as by intraperitoneal administration, intramuscular administration, or subcutaneous administration.
50. The use according to item 49, wherein said bath administration takes place for a time period of from 1 second to 5 hours, such as from 1 second to 2 hours, such as from 1 second to 1 hour, such as from 30 seconds to 1 hour or from 1 minute to 30 minutes.
51. The use according to any one of items 27-50, wherein the total concentration of bacteria in the probiotic composition is from 10<2 >to 10<14>, such as from 10<8 >to 10<13>, from 10<9 >to 10<13>, from 10<10 >to 10<12>, from 10<2 >to 10<7>, or from 10<5 >to 10<7>.
52. The use according to any one of items 27-51, wherein said probiotic composition is administered at least 10 days, such as at least 15 days, 20 days, 25 days, 30 days, or 40 days, before exposure of the teleost to said sea lice.
53. A method for treating and/or preventing a sea lice infestation in a teleost, said method comprising administering a therapeutically effective amount of a probiotic composition comprising probiotic bacteria of one or both of the species Aliivibrio njordis and Aliivibrio balderis to a teleost in need thereof.
54. The method according to item 53, wherein said probiotic composition comprises probiotic bacteria of both Aliivibrio njordis and Aliivibrio balderis.
55. The method according to item 53 or 54, wherein said Aliivibrio njordis is Aliivibrio njordis strain B1-25, 18-1/2013 mandib V11, which has been deposited at National Collection of Industrial and Marine Bacteria and has been assigned accession number NCIMB 42593.
56. The method according to any one items 53-55, wherein said Aliivibrio balderis is Aliivibrio balderis B1-24, 18-1/2013 kidn V12, which has been deposited at National Collection of Industrial and Marine Bacteria and has been assigned accession number NCIMB 42592.
57. The method according to any one items 53-56, wherein said probiotic composition further comprises probiotic bacteria of the species Aliivibrio nannie, such as Aliivibrio nannie B8-24, 313/2013 kidn V13, which has been deposited at National Collection of Industrial and Marine Bacteria and has been assigned accession number NCIMB 42594.
58. The method according to any one items 53-57, wherein said probiotic composition further comprises probiotic bacteria of the species Psychrobacter piscimesodermis, Psychrobacter piscimesenchymalis, Psychrobacter piscisubcutanea, Pseudomonas salmosubcutaneae, Pseudomonas salmosubpectoralis and/or Pseudomonas salmointermuscularis.
59. The method according to item 58, wherein said Psychrobacter piscimesodermis is Psychrobacter piscimesodermis Fisk 1, 41, 6/3-2014 Atl. Salm, which has been deposited at National Collection of Industrial and Marine Bacteria and has been assigned accession number NCIMB 42947.
60. The method according to item 58 or 59, wherein said Psychrobacter piscimesenchymalis is Psychrobacter piscimesenchymalis Fisk 2, 42, 6/3-2014 Atl. salm, which has been deposited at National Collection of Industrial and Marine Bacteria and has been assigned accession number NCIMB 42948.
61. The method according to any one of items 58-60, wherein said Psychrobacter piscisubcutanea is Psychrobacter piscisubcutanea Fisk 3, 43, 6/3-2014 Atl. Salm, which has been deposited at National Collection of Industrial and Marine Bacteria and has been assigned accession number NCIMB 42949.
62. The method according to any one of items 58-61, wherein said Pseudomonas salmosubcutaneae is Pseudomonas salmosubcutaneae Fisk 3, 13/5-2014, hb, Atl. Salm, which has been deposited at National Collection of Industrial and Marine Bacteria and has been assigned accession number 43330.
63. The method according to any one items 58-62, wherein said Pseudomonas salmosubpectoralis is Pseudomonas salmosubpectoralis Fisk 3, 13/5-2014, ba, Atl. salm., which has been deposited at National Collection of Industrial and Marine Bacteria and has been assigned accession number 43331.
64. The method according to any one of items 58-63, wherein said Pseudomonas salmointermuscularis is Pseudomonas salmointermuscularis Fisk 4, 13/5-2014, ha, Atl. salm., which has been deposited at National Collection of Industrial and Marine Bacteria and has been assigned accession number 43332.
65. The method according to any one of items 53-64, said method comprising administering a probiotic composition as defined in item 1 or 2 to said teleost before, during or after administration of one or more further probiotic composition(s) comprising one or more of a probiotic bacterium as defined in any one of items 3-12.
66. The method according to any one items 53-65, wherein said probiotic composition comprises probiotic bacteria of two or more different strains of the same bacterial species.
67. The method according to any one items 53-66, wherein said teleost is a marine teleost.
68. The method according to any one items 53-66, wherein said teleost is a fresh water teleost.
69. The method according to any one items 53-68, wherein said teleost is of the family Salmonidae, such as salmon, trout, and chars.
70. The method according to item 69, wherein said teleost of the family Salmonidae is Atlantic salmon (Salmo salar), brown trout (sea trout: Salmo trutta), Arctic char (Salvelinus alpinus), and/or any species of Pacific salmon, such as (Coho (Silver) salmon (Oncorhynchus kisutch), Chinook (King) (Oncorhynchus tshawytscha), pink (Humpy) salmon (Oncorhynchus gorbuscha), chum (Dog) salmon (Oncorhynchus keta), sockeye (Oncorhynchus nerka), cutthroat (Oncorhynchus clarki clarki) and steelhead (rainbow trout) (Oncorhynchus mykiss).
71. The method according to any one items 53-68, wherein said teleost is Atlantic cod, cleaner fish like lump fish and wrasses and freshwater fish as carps and perch. 72. The method according to item 71, wherein the sea lice is of the species Lepeophtheirus salmonis, Caligus C. rogercresseyi and/or Caligus elongatus. 73. The method according to any one items 53-72, wherein said probiotic composition further comprises a pharmaceutically acceptable excipient and/or adjuvant.
74. The method according to any one items 53-73, wherein said probiotic composition is administered in combination with a chemical parasitic infection treatment agent, such as an organophosphate, a carbamate, a pyrethroid, a pyrethrin, a synergist, an insect growth regulating chemical and/or an avermectin.
75. The method according to any one items 53-74, wherein said probiotic composition is administered via bath or dip administration in salt, fresh or brackish water, by oral administration, by oral administration or by injection, such as by intraperitoneal administration, intramuscular administration, or subcutaneous administration.
76. The method according to item 75, wherein said bath administration takes place for a time period of from 1 second to 5 hours, such as from 1 second to 2 hours, such as from 1 second to 1 hour, such as from 30 seconds to 1 hour or from 1 minute to 30 minutes.
77. The method according to any one of items 53-76, wherein the total concentration of bacteria in the probiotic composition is from 10<2 >to 10<14>, such as from 10<8 >to 10<13>, from 10<9 >to 10<13>, from 10<10 >to 10<12>, from 10<2 >to 10<7>, or from 10<5 >to 10<7>.
78. The method according to any one of items 53-77, wherein said probiotic composition is administered at least 10 days, such as at least 15 days, 20 days, 25 days, 30 days, or 40 days, before exposure of the teleost to said sea lice.

Claims (13)

1. A probiotic composition comprising probiotic bacteria of the species Aliivibrio njordis and/or Aliivibrio balderis, and optionally Aliivibrio nannie, for use in the treatment and/or prevention of a sea lice infestation in a teleost.
2. The probiotic composition for use according to claim 1, wherein said probiotic composition further comprises probiotic bacteria of the species Psychrobacter piscimesodermis, Psychrobacter piscimesenchymalis, Psychrobacter piscisubcutanea, Pseudomonas salmosubcutaneae, Pseudomonas salmosubpectoralis and/or Pseudomonas salmointermuscularis.
3. The probiotic composition for use according to claim 1 or 2, wherein said probiotic composition is administered before, during or after administration of one or more further probiotic composition(s) comprising one or more of a probiotic bacterium as defined in claim 2.
4. The probiotic composition for use according to any one of claims 1-3, wherein said probiotic composition comprises probiotic bacteria of two or more different strains of the same bacterial species.
5. The probiotic composition for use according to any one of the preceding claims, wherein said teleost is a marine teleost.
6. The probiotic composition for use according to any one of claims 1-4, wherein said teleost is a fresh water teleost.
7. The probiotic composition for use according to any one of the preceding claims, wherein said teleost is of the family Salmonidae, such as salmon, trout, and chars.
8. The probiotic composition for use according to claim 7, wherein said teleost of the family Salmonidae is Atlantic salmon (Salmo salar), brown trout (sea trout: Salmo trutta), Arctic char (Salvelinus alpinus), and/or any species of Pacific salmon, such as (Coho (Silver) salmon (Oncorhynchus kisutch), Chinook (King) (Oncorhynchus tshawytscha), pink (Humpy) salmon (Oncorhynchus gorbuscha), chum (Dog) salmon (Oncorhynchus keta), sockeye (Oncorhynchus nerka), cutthroat (Oncorhynchus clarki clarki) and steelhead (rainbow trout) (Oncorhynchus mykiss).
9. The probiotic composition for use according to any one of claims 1-6, wherein said teleost is Atlantic cod, cleaner fish like lump fish and wrasses and freshwater fish as carps and perch.
10. The probiotic composition for use according to any one of the preceding claims, wherein said sea lice is of the species Lepeophtheirus salmonis, Caligus rogercresseyi and/or Caligus elongatus.
11. The probiotic composition for use according to any one of the preceding claims, wherein said probiotic composition is administered in combination with a chemical parasitic infection treatment agent, such as an organophosphate, a carbamate, a pyrethroid, a pyrethrin, a synergist, an insect growth regulating chemical and/or an avermectin.
12. The probiotic composition for use according to any one of the preceding claims, wherein said probiotic composition is administered via bath or dip administration in salt, fresh or brackish water, by oral administration, or injection, such as bath administration for a time period of from 1 second to 5 hours, such as from 1 second to 2 hours, such as from 1 seconds to 1 hour, such as from 30 seconds to 1 hour or 1 minute to 30 minutes.
13. The probiotic composition for use according to any one of the preceding claims, wherein said probiotic composition is administered at least 10 days, such as at least 15 days, 20 days, 25 days, 30 days, or 40 days, before exposure of the teleost to said sea lice.
NO20191194A 2019-10-05 2019-10-05 Method for treating and/or preventing a sea lice infection in a teleost NO346319B1 (en)

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NO20191194A NO346319B1 (en) 2019-10-05 2019-10-05 Method for treating and/or preventing a sea lice infection in a teleost
EP20792930.8A EP4037698B1 (en) 2019-10-05 2020-10-02 A probiotic composition comprising probiotic bacteria for use in a method of treating and/or preventing a parasitic infection in a teleost
PCT/EP2020/077734 WO2021064217A1 (en) 2019-10-05 2020-10-02 Method for treating and/or preventing a parasitic infection in a teleost
AU2020357769A AU2020357769A1 (en) 2019-10-05 2020-10-02 Method for treating and/or preventing a parasitic infection in a teleost
CA3156807A CA3156807A1 (en) 2019-10-05 2020-10-02 Method for treating and/or preventing a parasitic infection in a teleost
US17/766,551 US20230338437A1 (en) 2019-10-05 2020-10-02 Method for treating and/or preventing a parasitic infection in a teleost
CL2022000819A CL2022000819A1 (en) 2019-10-05 2022-04-01 Method for the treatment and/or prevention of a parasitic infection in a teleost
DKPA202200420A DK202200420A1 (en) 2019-10-05 2022-05-04 Method for treating and/or preventing a parasitic infection in a teleost

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015155293A1 (en) * 2014-04-09 2015-10-15 Biomar Group A/S Compound or composition for use in the prevention and/or treatment of an ectoparasitic copepod infestation or infection in fish
WO2018007632A1 (en) * 2016-07-08 2018-01-11 Previwo As Probiotic bacteria for fish
WO2019135009A1 (en) * 2018-01-08 2019-07-11 Previwo As Probiotic bacteria for fish

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015155293A1 (en) * 2014-04-09 2015-10-15 Biomar Group A/S Compound or composition for use in the prevention and/or treatment of an ectoparasitic copepod infestation or infection in fish
WO2018007632A1 (en) * 2016-07-08 2018-01-11 Previwo As Probiotic bacteria for fish
WO2019135009A1 (en) * 2018-01-08 2019-07-11 Previwo As Probiotic bacteria for fish

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