WO2020130843A1 - Composition destinée à éliminer les crustacés ectoparasites des salmonidés d'élevage à l'aide de la curcumine ou d'un dérivé pharmaceutiquement acceptable de celle-ci et de la lumière - Google Patents

Composition destinée à éliminer les crustacés ectoparasites des salmonidés d'élevage à l'aide de la curcumine ou d'un dérivé pharmaceutiquement acceptable de celle-ci et de la lumière Download PDF

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Publication number
WO2020130843A1
WO2020130843A1 PCT/NO2019/050279 NO2019050279W WO2020130843A1 WO 2020130843 A1 WO2020130843 A1 WO 2020130843A1 NO 2019050279 W NO2019050279 W NO 2019050279W WO 2020130843 A1 WO2020130843 A1 WO 2020130843A1
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Prior art keywords
light
treatment
curcumin
lice
salmon
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PCT/NO2019/050279
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English (en)
Inventor
Øyvind OALAND
Jo Klaveness
Gordon Ritchie
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Mowi ASA
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Publication date
Application filed by Mowi ASA filed Critical Mowi ASA
Priority to CA3123798A priority Critical patent/CA3123798A1/fr
Priority to EP19901125.5A priority patent/EP3897114A4/fr
Publication of WO2020130843A1 publication Critical patent/WO2020130843A1/fr
Priority to DKPA202170358A priority patent/DK202170358A1/en

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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
    • A01P7/00—Arthropodicides
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K61/00—Culture of aquatic animals
    • A01K61/10—Culture of aquatic animals of fish
    • A01K61/13—Prevention or treatment of fish diseases
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K61/00—Culture of aquatic animals
    • A01K61/60—Floating cultivation devices, e.g. rafts or floating fish-farms
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K63/00—Receptacles for live fish, e.g. aquaria; Terraria
    • A01K63/04—Arrangements for treating water specially adapted to receptacles for live fish
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K63/00—Receptacles for live fish, e.g. aquaria; Terraria
    • A01K63/06—Arrangements for heating or lighting in, or attached to, receptacles for live fish
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
    • A01N25/002—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing a foodstuff as carrier or diluent, i.e. baits
    • A01N25/006—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing a foodstuff as carrier or diluent, i.e. baits insecticidal
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N27/00—Biocides, pest repellants or attractants, or plant growth regulators containing hydrocarbons
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N35/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having two bonds to hetero atoms with at the most one bond to halogen, e.g. aldehyde radical
    • A01N35/02—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having two bonds to hetero atoms with at the most one bond to halogen, e.g. aldehyde radical containing aliphatically bound aldehyde or keto groups, or thio analogues thereof; Derivatives thereof, e.g. acetals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/12—Ketones
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • A61K31/716—Glucans
    • A61K31/724—Cyclodextrins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/88—Liliopsida (monocotyledons)
    • A61K36/906—Zingiberaceae (Ginger family)
    • A61K36/9066—Curcuma, e.g. common turmeric, East Indian arrowroot or mango ginger
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
    • A61K41/0057—Photodynamic therapy with a photosensitizer, i.e. agent able to produce reactive oxygen species upon exposure to light or radiation, e.g. UV or visible light; photocleavage of nucleic acids with an agent
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61N—ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY
    • A61N5/00—Radiation therapy
    • A61N5/06—Radiation therapy using light
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61N—ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY
    • A61N5/00—Radiation therapy
    • A61N5/06—Radiation therapy using light
    • A61N5/0613—Apparatus adapted for a specific treatment
    • A61N5/062—Photodynamic therapy, i.e. excitation of an agent
    • Y—GENERAL 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
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
    • Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
    • Y02A40/81—Aquaculture, e.g. of fish

Definitions

  • the invention relates to a new method for removing or killing of ectoparasites from
  • the invention relates to removing or killing crustacean
  • the fish may be a salmonid fish, such as Atlantic
  • the crustacean ec toparasite may be salmon lice ( Lepeophtheirus salmonis, Caligus rogercresseyi, Ca- ligus spp.J. Even more particularly the invention relates to a method comprising
  • io treatment means where light from an artificial light source is used in combination with
  • the method relates to the use of a combina tion of regulatory approved food additives or cyclodextrin complexes thereof and light.
  • Aquaculture also referred to as aquafarming, is the farming of aqueous organism.
  • Aquaculture involves cultivating freshwater and saltwater populations under controlled conditions and is in contrast to commercial harvesting or fishing where the organisms are naturally present.
  • the farmed organisms can typically be fish, crustaceans, mol- lusks, aquatic plants, algae.
  • the aquaculture farms can be in the form of tanks (closed 20 or semi closed), fish ponds, ocean cages or nets.
  • Photodynamic therapy is generally a term for a therapy that involves a so-called pho tosensitizer and light, and oxygen, where the light activates the photosensitizer to generate singlet oxygen and other reactive and toxic species.
  • Photodynamic therapy is well described in human medicine, see for example: Kwiatkowski S et al. Photodyna- mic therapy - mechanisms, photosensitizers and combinations. Biomed Pharmacother. 2018 Oct; 106: 1098-1107.
  • the clinical use of photodynamic therapy is, however, lim ited to some specific indications; especially some skin diseases with products compris ing 5-aminolevulinic acid methyl ester as photosensitizer, and age-related macular degeneration with products comprising verteporfin as photosensitizer.
  • Very many of the publications on photodynamic therapy relates to cancer and some publications relate to infections, however, these diseases are not commonly treated by photody namic therapy.
  • the photosensitizer used in these studies is a porphyrin derivative.
  • Methylene blue is a cationic (permanent positively charged) synthetic compound and nuclear fast red is a negatively charged sulphonic acid in sea water (pH 7.5 to 8.4).
  • Acartia clausi is a small animal (1 mm in length, 0.2 mm broad). The length of salmon lice is up to 18 mm.
  • Hydrogen peroxide has for many years been used to combat sea lice infections in fish including salmon.
  • the concentrates are diluted to a final concentration of 1500 mg hydrogen peroxide per liter.
  • Hydrogen peroxide is toxic for salmon lice.
  • the mecha nism of action is probably related to an oxidative effect on lice components and the formation of oxygen emboli within the salmon lice.
  • Drug resistance is a general problem with farming of salmon. See for example Stian Morch Aaen et al. Drug Resistance in sea lice: a threat to salmon aquaculture.
  • the invention relates to a new method for treatment of salmon lice (L. salmonis, C. rogercresseyi , Caligus spp.) in farmed fish such as Atlantic salmon (S. salar L.) using an artificial light source and at least one chemical substance at toxicologically ac ceptable concentrations for Atlantic salmon. More specifically, the present inventors have identified a novel therapy for the treatment of salmon louse in farmed Atlantic salmon populations that comprises a combination of artificial light together with regu latory approved food additives.
  • the fish may be a salmonid fish, such as Atlantic salmon ( Salmo salar L.) or rainbow trout ( Oncorhynchus mykiss).
  • Atlantic salmon Salmo salar L.
  • rainbow trout Oncorhynchus mykiss
  • Atlantic salmon for convenience fish is referred to as Atlantic salmon in the preceding text and in the text to follow, however, without excluding rainbow trout, other salmonid species or other relevant fish species.
  • blue light comprises violet light (380nm to 450 nm, blue light (450nm to 485nm) and cyan light (485nm to 500nm).
  • Blue light as used herein covers the wavelength from 380nm to 500nm.
  • Red light as used herein covers the wavelength from 625nm to 740nm.
  • the invention relates in a first aspect to a composition in the form of an aqueous solution comprising a photosensitizer for use in a method of photodynamic therapy for an external crustacean parasite infection in salmonid fish, said external crustacean parasite infection comprises an infection of salmon lice, Lepeophtheirus salmonis, Caligus rogercresseyi and Caligus spp.
  • Said photosensitizer is curcumin or a pharmaceutical acceptable derivative thereof, and said photosensitizer is administered in a bath treatment of the salmonid fish in need of such treatment, and the salmon lice is illuminated by light.
  • said photosensitizer may be a curcumin cyclodextrin.
  • said light may be blue light. In an alternative embodiment said light may be red light.
  • the salmonid fish may be located in a receptacle.
  • the receptacle may be one of a closed tank, a semi-closed tank, a chamber, a container and a fish pond.
  • the salmonid fish may be located in one of an ocean cage and a net pen.
  • the blue light source may comprise of LED lamps.
  • the LED lamps may emit light of a wavelength of 450nm to 460nm.
  • Said photosensitizer may be added in the water surrounding the salmonid fish.
  • the water surrounding the salmonid fish may form the bath treatment.
  • the invention relates to use of an artificial light device emitting blue light in accordance to the first aspect of the invention.
  • the invention may relate to a composition in the form of an aqueous solution comprising a photosensitizer for use in a method of photodynamic therapy for an external crustacean parasite infection in salmonid fish, said external crustacean parasite infection comprises an infection of salmon lice, Lepeophtheirus salmonis, Ca- ligus rogercresseyi and Caligus spp.
  • Said photosensitizer is riboflavin or a pharmaceu tical acceptable derivative thereof, and said photosensitizer is administered in a bath treatment of the salmonid fish in need of such treatment, and the salmon lice is illu minated by light.
  • said photosensitizer may be a riboflavin phosphate.
  • said light may be blue light. In an alternative embodiment said light may be red light.
  • the salmonid fish may be located in a receptacle.
  • the receptacle may be one of a closed tank, a semi-closed tank, a chamber, a container and a fish pond.
  • the salmonid fish may be located in one of an ocean cage and a net pen.
  • the blue light source may comprise of LED lamps.
  • the LED lamps may emit light of a wavelength of 450nm to 460nm.
  • Said photosensitizer may be added in the water surrounding the salmonid fish.
  • the water surrounding the salmonid fish may form the bath treatment.
  • the invention may relate to a composition in the form of an aqueous solution comprising a photosensitizer for use in a method of photodynamic therapy for an external crustacean parasite infection in salmonid fish, said external crustacean parasite infection comprises an infection of salmon lice, Lepeophtheirus salmonis, Ca- ligus rogercresseyi and Caligus spp.
  • Said photosensitizer is chlorophyllin sodium cop per salt or a pharmaceutical acceptable salt or derivative thereof, and said photosensi tizer is administered in a bath treatment of the salmonid fish in need of such treatment, and the salmon lice is illuminated by light.
  • said light may be blue light. In an alternative embodiment said light may be red light.
  • the salmonid fish may be located in a receptacle.
  • the receptacle may be one of a closed tank, a semi-closed tank, a chamber, a container and a fish pond.
  • the salmonid fish may be located in one of an ocean cage and a net pen.
  • the blue light source may comprise of LED lamps.
  • the LED lamps may emit light of a wavelength of 450nm to 460nm.
  • Said photosensitizer may be added in the water surrounding the salmonid fish.
  • the water surrounding the salmonid fish may form the bath treatment.
  • the energy required for the artificial light source according to the present invention can vary from a few watts to several kilowatts depending upon the nature of the artifi cial light source and the volume of the enclosure or ocean cage or net.
  • the intensity of the light diminishes with the distance between the light source and the target surface, e.g. the surface of the sea lice.
  • the light intensity may be too high at a too close distance, leading to a too rapid bleaching of the photosensitizer, and thereby the photosensitizer lose its effect.
  • the light intensity may be too low at a too far distance, leading to that photosensitizer is not activated. It is therefore within the scope of the present invention to optimize the distance between the light source and the target surface.
  • the optimal distance is thus dependent on among other things the actual wavelength, the actual photosensitizer, and the actual specification of the lamp, e.g. output in lu men.
  • the exact mechanism of the combination of light together with the regulatory ap proved food additives on salmon lice is not studied, but it can be speculated that the light activates the regulatory approved food additives through a photochemical pro cess and that the formed reactive species are responsible for the observed increased toxicity on the salmon lice.
  • Said photosensitizer may comprise a regulatory approved food additive.
  • Said photo sensitizer may be a food additive cyclodextrin complex where the food additive com prises the regulatory approved food additive.
  • the regulatory approved food additive may be at least one of E100 Curcumin, E101(i) Riboflavin, E101(ii) Riboflavin-5'-phosphate, E102 Tartrazine, E123 Amaranth, E127 Erythrosine, E129 Allura Red AC, E131 Patent Blue V, E132 Indigotine; Indigo Car mine, E133 Brilliant Blue FCF, E140 Chlorophylls and chlorophyllins, E141 Copper complexes of chlorophyll and chlorophyllins, E142 Green S, E151 Brilliant Black BN; Black PN, E155 Brown HT, E160a Carotenes, E160b Annatto; Bixin; Norbixin, E160c Paprika extract; Capsanthian; Capsorubin, E160d Lycopene, E160e Beta-apo-8'- carotenal (C30), E161b Lutein, E161g Canthaxanthin, E162 Beetroot Red; Betanin,
  • the artificial light source may comprise of LED lamps.
  • the wavelength of the emitted light from the artificial light source may be between 200 and 800 nm.
  • the wavelength of the emitted light may be between 400 and 800 nm.
  • the wavelength of the emitted light may be between 200 and 400 nm.
  • the wavelength of the emitted light may be between 200 and 300 nm.
  • Said artificial light source may be a part of a device for automatic localization of salmon lice.
  • Said artificial light source may be a light source used in cultivation or farming of salmonid fish.
  • Said photosensitizer may be added in a food pellet for feeding the salmonid fish.
  • Said photosensitizer may be added in the water surrounding the salmonid fish.
  • An artificial light device may be used according to the methods described above.
  • the present invention may relate to a treatment of salmon louse in farmed fish populations that comprises a combination of regulatory approved food additives as cyclodextrin complexes and artificial light.
  • the present invention may relate to a protocol for the present treat ment.
  • the food additive is administered prior to, after or at approximately the same time as the artificial light.
  • the present invention may relate to a protocol for the present treat ment.
  • the present treatment can be performed when the Atlantic salmon is in recep tacles or enclosures, e.g. closed tanks or semi-closed tanks, pipes, chambers, con tainers, fish ponds, ocean cages or net pens or a combination of such enclosures.
  • the present invention may relate to the artificial light source.
  • the light source according to the present invention can be a light source positioned in the air or placed in water.
  • the light source can optionally be a plurality of light sources.
  • the light source should generate light with wave lengths from 200 to 800 nm.
  • the light source might also generate electromagnetic radiation that is outside this range.
  • the light source might be in the form of a laser.
  • the light source or light systems may comprise halogen lamps, mercury lamps, light emitting electrodes (LEDs) or other suitable light systems.
  • the light source can be a stationary light source or a movable light source.
  • the present invention may relate to a drug treatment protocol.
  • other drug substances or drug like compounds might, in addition to regulatory approved food additives, be useful for the treatment according to the present invention.
  • These drugs include other drugs that have been shown to be toxic to salmon lice including commercial products that are approved for salmon lice infections in Atlantic salmon.
  • the present invention may relate to the dose or concentration of the regulatory approved food additive in the clinical situation.
  • the present invention may relate to the exposure time of the treatment process.
  • the time necessary to perform the present process varies from minutes to hours depending upon several factors like for example nature of the salmon lice infec tion, number of Atlantic salmon per cubic meter of water, concentration of regulatory approved food additive in the water and optionally other drugs and drug like com pounds and nature of the artificial light source.
  • the present invention may relate to resistance.
  • the method according to the present invention is new for treatment of Atlantic salmon infected by salmon lice and has ad vantages related to treatment of Atlantic salmon infected by resistant salmon lice and also related to generation of new forms of resistance.
  • the possible mechanism of ac- tion related to the present invention is biologically not a typical process where re sistance processes are generated.
  • the advantages by a combination of light together with food additives according to the present invention versus state-of-the-art treatment of salmon lice relate to one or more of the following topics: reduced amount of drugs, improved efficacy on salmon lice, reduced toxicity to the Atlantic salmon, resistance issues, cost and time.
  • the present invention relates to the selection of food additives. Approved food addi tives are listed with E-numbers. See for example: https://www.food.aov.uk/business- auidance/eu-approved-additives-and-e-numbers
  • the preferred compounds according to the present invention are:
  • E100 Curcumin E101(i) Riboflavin; (ii) Riboflavin-5'-phosphate, E102 Tartrazine, E123 Amaranth, E127 Erythrosine, E129 Allura Red AC, E131 Patent Blue V, E132 Indigo- tine; Indigo Carmine, E133 Brilliant Blue FCF, E140 Chlorophylls and chlorophyllins, E141 Copper complexes of chlorophyll and chlorophyllins, E142 Green S, E151 Brilliant Black BN; Black PN; E155 Brown HT, E160a Carotenes, E160b Annatto; Bixin; Norbix in, E160c Paprika extract; Capsanthian; Capsorubin, E160d Lycopene, E160e Beta- apo-8'-carotenal (C30), E161b Lutein, E161g Canthaxanthin, E162 Beetroot Red; Betanin, E163 Anthocyanins and E180
  • E100 Curcumin E101(i) Riboflavin; (ii) Riboflavin-5'-phosphate, E102 Tartrazine, E123 Amaranth ,E127 Erythrosine ,E129 Allura Red AC,E131 Patent Blue V, E132 Indigotine; Indigo Carmine ,E133 Brilliant Blue FCF, E140 Chlorophylls and chlorophyllins, E141 Copper complexes of chlorophyll and chlorophyllins, E142 Green S, E160a Carotenes E160c Paprika extract; Capsanthian; Capsorubin, E160d Lycopene, E161b Lutein, E161g Canthaxanthin, E162 Beetroot Red; Betanin and E163 Anthocyanins.
  • the even most preferred compound is curcumin.
  • the concentration of the food additive varies depending upon the various parameters like for example choice of food additive, temperature, disease stage, light source and the density of Atlantic salmon.
  • Typical concentration range for the food additives ac cording to the present invention is in the range 0.1 micromolar (1 time 10 7 molar (M)) to 400 micromolar (400 times 10 6 molar(M)). More preferred the concentration range for the food additive cyclodextrin complex according to the present invention is in the range 0.5 micromolar to 300 micromolar.
  • the present invention relates to treatment of salmon louse in farmed fish populations that comprises a combination of regulatory approved food additives as cyclodextrin complexes and light from an artificial light source.
  • Cyclodextrins are cyclic oligosaccharides what are known to form complexes with various chemical and drugs. Drug complexes with CDs are currently used in hu man medicine. The potential advantages by using drug CD complexes relate to chemi cal stability, aqueous solubility and oral bioavailability.
  • CDs include alpha-CD which is a 6-membered sugar ring molecule, beta-CD which is a 7-membered sugar ring molecule, gamma-CD which is a 7-membered sugar ring molecule and an almost unlimited number of chemical deriva tives with various degree of substitution on these three different sugar ring molecules.
  • the preferred and the most preferred food additives also relate to the food additive cyclodextrin complexes.
  • cyclodextrin part of the food additive cyclodextrin complexes are the three unsubstituted cyclodextrins, methyl cyclodextrin and 2-hydroxypropyl cyclodex trins the preferred cyclodextrins.
  • cyclodextrin part of the food additive cyclodextrin complexes is beta- cyclodextrin the most preferred cyclodextrin.
  • curcumin beta-cyclodextrin, chlorophyll beta-cyclodextrin and chlorophyllin beta-cyclodextrin are among the most preferred food additive cy clodextrin complexes to be used together with artificial light for treatment of salmon louse in farmed fish according to the present invention.
  • the food additive cyclodextrin complexes according to the present invention can be prepared by standard methods for preparation of such complexes. These methods in clude co-evaporation, spray-drying, freeze drying and kneading.
  • One simple method is to carefully mix the food additive with the cyclodextrin (typically molar ratio 1 to 5) with small amounts of water forming a very thick paste for 10 minutes using a mortar and pestle, dry the paste in an oven and prepare a powder of the dry material using the mortar and pestle.
  • the concentration of the food additive cyclodextrin complex varies depending upon the various parameters like for example choice of food additive, temperature, disease stage, light source and the density of Atlantic salmon.
  • Typical concentration range for the food additive cyclodextrin complex according to the present invention is in the range 0.1 micromolar (1 time 10 7 molar (M)) to 400 micromolar (400 times 10 6 mo- lar(M)). More preferred the concentration range for the food additive cyclodextrin complex according to the present invention is in the range 0.5 micromolar to 300 mi cromolar.
  • the present invention relates to a protocol for the present treatment.
  • food additive is administered prior to, after or at approximately the same time as the light.
  • the light might be present after the food additive concentration around the Atlantic salmon is reduced. Any protocol using food additive and light is within the scope of the present invention. It is up to the skilled person on salmon lice treatment to select the best suited protocol for each treatment.
  • a first preferred protocol is to apply the regulatory approved food additive some time prior to the light treatment.
  • the light and the regulatory approved food additive are present partly together or the light is present only after the aqueous con centration of hydrogen peroxide is reduced but still present within the salmon louse.
  • a second preferred protocol is to apply the regulatory approved food additive and to gether and optionally continue to use light after the aqueous concentration of the reg ulatory approved food additive is reduced.
  • a third preferred protocol is to apply light after the aqueous concentration of the regu latory approved food additive is reduced but the regulatory approved food additive still is present within the salmon lice.
  • the most preferred protocol is to apply light, at least partly, when the concentration of the regulatory approved food additive still is quite high in the water surrounding the Atlantic salmon and the salmon lice.
  • the present invention relates to a protocol for the present treatment.
  • the present treatment can be performed when the Atlantic salmon is in receptacles or enclosures, e.g. closed tanks or semi-closed tanks, pipes, chambers, containers, fish ponds, ocean cages or net pens or a combination of such enclosures.
  • the preferred methods of the present invention relate to treatment of Atlantic salmon with salmon lice within a tank/boat or ocean cages or net pens.
  • the absolutely most preferred methods of the present invention relate to treatment of Atlantic salmon infected with salmon lice within an ocean cage or a net pen.
  • the present invention relates to the position of the artificial light source.
  • One preferred position of the light source is in the air above the water.
  • Another preferred position of the light source is on the walls or the bottom of a receptacle or enclosure, e.g. closed tank or semi-closed tank, pipe, chamber, container, or fish ponds.
  • Another preferred position of the light source is under water within the receptacle or the ocean cage or net by mechanical arrangements and/or floating devices. The most preferred position of the light source in a large ocean cage or net is under water.
  • the artificial light source should generate light that has wave lengths from 200nm to 800nm.
  • One preferred artificial light source according to the present invention is that the artificial light source generates visible light within the wavelength band from 400nm to 800nm.
  • One preferred artificial light source according to the present invention is that the artifi cial light source generates light within one of the following colors: red, yellow, green, blue or white.
  • Another preferred light source according to the present invention is that the artificial light source generates UV light within the wavelength band from 200nm to 400nm.
  • Another preferred artificial light source according to the present invention is that the artificial light source generates UVC light within the wavelength band from 200nm to 300nm.
  • the artificial light source might also generate electromagnetic radiation that is outside these wavelength bands. If so, the artificial light source must also generate some light within referred wave lengths or colors.
  • the artificial light source can be one single artificial light source or a plurality of artifi cial light sources. If the volume is huge, like in an ocean cage or net, it is according to the present invention preferred to use a plurality of artificial light sources.
  • One artificial preferred light source according to the present invention is that the artifi cial light source is based on laser.
  • the artificial light source is based on LEDs, halogen lamp or mercury lamp.
  • the most pre ferred type of lamps among these lamps are LED lamps.
  • the most preferred artificial light sources according to the present invention are LED lamps generating blue light; typically, in the wavelength range from 380nm to 500nm. Examples of such lamps are optionally dimmable, underwater lamps currently com suddenly available and used in aquaculture for other purposes.
  • the light from the artificial light source may be stationary, i.e. same light in the same area over time, or the artificial light source may be movable.
  • the present invention relates to a drug treatment protocol.
  • other drug substances or drug like compounds may, in addition to the regulatory approved food additive, be useful for the treatment according to the present invention.
  • One preferred treatment protocol according to the present invention is that the other drug substances or drug like compounds are selected among other drugs that are reg ulatory approved for use to treat Atlantic salmon infected by salmon lice.
  • Another preferred treatment protocol according to the present invention is that the other drug substances or drug like compounds that are cholinesterase inhibitors, syn thetic pyrethroids, chitin synthase inhibitors or glutamate-based chlorine ion channel regulators.
  • Another preferred treatment protocol according to the present invention is that the other drug substances or drug like compounds are selected among compounds that are photosensitizers.
  • Another preferred treatment protocol according to the present invention is that the other drug substances or drug like compounds are selected among compounds that are phototoxic.
  • the present invention relates to the dose or concentration of the regulatory approved food additive in the clinical situation.
  • the present invention relates to the timing of the treatment process.
  • the time neces- sary to perform the present process varies from minutes to hours depending upon several factors like for example nature of the salmon lice infection, number of Atlantic salmons per cubic meter, the nature and the concentration of the regulatory approved food additive and optionally other drugs and drug like compounds, and nature of the light source.
  • the present invention relates to resistance.
  • the method according to the present in vention is new for treatment of Atlantic salmon infected by salmon lice and has ad vantages related to treatment of Atlantic salmon infected by resistant salmon lice and also relates to generation of new forms of resistance.
  • the possible mechanism of ac tion related to the present invention is biologically not a typical process where re- sistance processes are generated.
  • the advantages by a combination of light together with the regulatory approved food additive according to the present invention relate to one or more of the following top ics: reduced amount of drugs, improved efficacy on salmon lice, and reduced toxicity to the Atlantic salmon.
  • Chlorophyllin sodium copper salt (1.412 gram, 2mmol) and 2-Hydroxypropyl-3- cyclodextrin (Average Mw ⁇ 1,460) (4.38 gram, 3mmol) were mixed in a mortar with a pestle. Water (3 ml) was added and the formed paste was mixed in the mortar for 5 minutes. The product was dried at 60 degrees centigrade in dark overnight. The prod uct was in the form of a green powder. The product comprised 24.4% chlorophyllin sodium copper salt.
  • Chlorophyllin sodium copper salt (1.412 gram, 2mmol) and gamma-cyclodextrin (3.89 gram, 3mmol) were mixed in a mortar with a pestle. Water (3 ml) was added and the formed paste was mixed in the mortar for 5 minutes. The product was dried at 60 degrees centigrade in dark overnight. The product was in the form of a green pow der. The product comprised 26.6% chlorophyllin sodium copper salt.
  • Curcumin (0.736 gram, 2mmol) and gamma-cyclodextrin (3.89 gram, 3mmol) were mixed in a mortar with a pestle. Water (5 ml) was added and the formed paste was mixed in the mortar for 5 minutes. The product was dried at 60 degrees centigrade in dark overnight. The product was in the form of a yellow powder. The product com prised 15.9 % curcumin.
  • Methylene Blue hydrate (640 mg, 2mmol) and 2-Hydroxypropyl-3-cyclodextrin (Aver age Mw ⁇ 1,460) (4.38 gram, 3mmol) were mixed in a mortar with a pestle. Water (3 ml) was added and the formed paste was mixed in the mortar for 5 minutes. The product was dried at 65 degrees centigrade in dark overnight. The product was in the form of a blue powder. The product comprised 12.8% methylene blue.
  • Methylene Blue hydrate (640 mg, 2mmol) and gamma-cyclodextrin (3.89 gram, 3mmol) were mixed in a mortar with a pestle. Water (3 ml) was added and the formed paste was mixed in the mortar for 5 minutes. The product was dried at 65 degrees centigrade in dark overnight. The product was in the form of a green powder. The product comprised 14.1% methylene blue.
  • Curcumin-8-cvclodextrin complex (1 :4)
  • Curcumin (0.368 gram, lmmol) and b-cyclodextrin (4.54 gram, 4mmol) were mixed in a mortar with a pestle. Water (5 ml) was added and the formed paste was mixed in the mortar for 5 minutes. The product was dried at 65 degrees centigrade in dark overnight. The product was in the form of a yellow powder. The product comprised 7.8% curcumin.
  • Curcumin (0.368 gram, lmmol), Polysorbate 80 (250mg) and b-cyclodextrin
  • the product was in the form of a yellow powder.
  • the product comprised 7.4% curcumin.
  • Curcumin-8-cvclodextrin complex (1 :4)
  • Curcumin-8-cvclodextrin complex (1 :4)
  • curcumin 0.368 gram, lmmol
  • Polysorbate 80 0.25g
  • b- cyclodextrin 9.08 gram, 8mmol
  • water 10 ml
  • Curcumin-8-cvclodextrin complex (1 :8)
  • curcumin 0.368 gram, lmmol
  • b-cyclodextrin 9.08 gram, 8mmol
  • water 10 ml
  • Salmon louse ( Lepeophtheirus salmonis ) were collected from a salmon farming facility in Norway.
  • Chlorophyllin sodium copper salt (Sigma C6003-25G), concentration in seawater: 4 mg per litre (chlorophyll solution; 5.5 micromolar).
  • 5-Aminolevulinic acid hydrochloride (Sigma-Aldrich). Concentration in sea- water: 168 mg per litre (5-ALA solution; 1 millimolar).
  • Group 1 White LED light (240V, 50W, 35001m)
  • Group 2 Red light. 100 red LED lamp array (10 x 10) mounted on a heat sink. Speci- fications: 620-625nm, DC32-36V, 3500mA, 1000-1500lm
  • each treatment received additional aeration throughout the exposure period. This provided aeration and ensures that the treatment solution remains in suspension, allowing for even distribution of the treatment compound.
  • Group 3 was placed in the darkened cabinet immediately following exposure to the treatment compound. Sixty minutes after the exposure period began a count of the number of affected/unaffected lice was conducted.
  • Groups 1 and 2 were exposed to treatment compound for 1 hour, after which the lights were turned on and they were then exposed to a combination of treatment compound and light source. One hour after the start of light exposure a count of the number of affected/unaffected lice was conducted.
  • the curcumin solution (Compound 3) resulted in 60% parasites affected by the treatment.
  • the chlorophyll solution (chlorophyllin sodium copper salt) treatment group showed no treatment ef fect, in fact, a number of egg string had hatched in the chlorophyll solution, all nauplii present were alive and swimming.
  • the 5-ALA solution showed no treatment effect.
  • the concentration of 5-ALA was 1 millimolar which was 50 times higher than the con centration of curcumin which was 20 micromolar.
  • Salmon louse ( Lepeophtheirus salmonis ) were collected from a salmon farming facility in Norway.
  • Treatment II Curcumin beta-cyclodextrin labelled MH-107 (1 :4) (Compound 7). Con- centration : 640 micromolar (22.7 mg curcumin (304 mg complex) per liter). Treatment III: Copper complex of chlorophyllin sodium salt 2-hydroxypropyl-beta- cyclodextrin complex (2:3) (Compound 1). Concentration: 22 micromolar (16 mg copper complex of chlorophyllin sodium salt (64 mg complex) per liter).
  • Red light 100 red LED lamp array (10 x 10) mounted on a heat sink. Specifications: 620-625nm, DC32-36V, 3500mA, 1000-1500lm). (The power supply did not function optimally in these experiments resulting in somewhat reduced intensity of the red light (only 30 V, 0.51 A)).
  • Blue light 100 blue LED lamp array (10 x 10) mounted on a heat sink. Specifications: 380-500 nm, DC32-36V, 3500mA (100W). Luminous flux: 5000-6000lm.
  • Eleven glass flasks were filled with 300 ml seawater. Ten lice were transferred, using forceps, into each flask. The flasks were randomly labelled and distributed among the 3 light groups. Due to limited lice numbers only 2 control flasks were available, so only 2 treatment groups had controls. Each flask was gently stirred to ensure that all lice were attached, the contents were emptied over a sieve, after which 300 ml treatment compound was then introduced into the flask. The flasks were then placed in their respective waterbath. Sixty minutes after the exposure period began a count of the number of affected/unaffected lice was conducted.
  • treatment I had 30% affected parasites as compared to the other treatments with no affected parasites.
  • the treatments were more effective at the 24 h count, with treatment I being most effective with 100% affected lice, treatment II had 80% affected lice, and treatment III had 30% affected lice (Table 4).
  • treatment I contained a lot of precipitate, whereas the other groups had none.
  • Treatment efficacy at the 1 h control was low with treatment I resulting in 20% affected lice and treatment III with 10% affected lice (Table 5). By the 24 h mark treatment III resulted in 0% affected lice.
  • treatment I resulted in 80% affected lice, and treatment II had 50% affected lice (Table 5). Again, there were issues with the presence of a high amount of precipitate in treatment I.
  • the blue LED group resulted in no affected lice in either the control or treatment III for both the 1 h and 24 h counts (Table 6).
  • Treatment I was the most effective with 70% affected lice at the 1 h count and 90% affected lice at the 24 h count.
  • Treatment II resulted in 20% affected lice after 1 h and 30% affected lice after 24 h of exposure. In the blue light group at the 24 h count, treatment III had large amounts of precipitate out of solution with lice observed moving through it (Table 5).
  • Salmon louse ( Lepeophtheirus salmonis ) were collected from a salmon farming facility in Norway. LABORATORY SETUP
  • Treatment I Curcumin 2-hydroxypropyl-beta-cyclodextrin (MH-103). Concentra tion : 160 micromolar curcumin (408 mg complex per liter). Treatment II (T2) : Riboflavin 5'-phosphate sodium Ph. Eur. Concentration : 100 mi cromolar (47.8 mg per liter).
  • Treatment III (T3) : Riboflavin 5 Ph. Eur. Concentration : 100 micromolar (37.6 mg per liter).
  • Treatment IV (T4) : Riboflavin 5 Ph. Eur. Concentration: 300 micromolar (112.8 mg per liter).
  • Treatment V T5 : Riboflavin 5'-phosphate sodium Ph. Eur. Concentration : 300 mi cromolar (143.4 mg per liter).
  • Blue light 100 blue LED lamp array (10 x 10) mounted on a heat sink. Specifications: 450-460nm, DC32-36V, 3500mA (100W), Luminous flux: 1000-1500lm).
  • curcumin cyclodextrin 160 micromolar was found to be potent. The percentage of active salmon lice after 40 minutes was zero using blue light. The treatment I solution was also active using white light with 50% active lice after 40 minutes and 20% active lice after 90 minutes.
  • the following lice strains were used. The first was sourced from iLab, Bergen, where they cultured, collected, and shipped the lice to the laboratory based at NMBU
  • the lice were shipped in a single 2.5L container filled with oxygenated seawater.
  • the second was sourced from the University of Bergen (UiB), where they cultured, collected, and shipped the lice to the laboratory based at NMBU.
  • the lice were shipped in four 2.5L plastic bottles, filled with oxygenated seawater. All surviving lice were placed in a temperature-controlled cabinet (12°C) with added air supply until commencement of the bioassays.
  • bioassays were conducted at a constant water temperature of 12°C using two water baths and a temperature-controlled cabinet and included the following three light treatments: white LED, blue LED and no light (Table 1). The bioassays were conducted in two rounds, one round for each parasite strain using the same treatment compounds being used as shown in Table 2.
  • Blue light 100 blue LED lamp array (10 x 10) mounted on a heat sink and cooled by an external fan. Specifications: 450-460nm, DC32-36V, 3500mA (100W), Luminous flux: 1000-1500lm).
  • Treatment II Riboflavin phosphate. Concentration : 300 micromolar Treatment III (T3) : Sea water control
  • Treatment IV 10 minutes: Curcumin 2-hydroxypropyl-beta-cyclodextrin.
  • Concen tration 160 micromolar 408 mg complex per liter
  • Treatment V (T5) : 20 minutes: Curcumin 2-hydroxypropyl-beta-cyclodextrin.
  • Concen tration 160 micromolar 408 mg complex per liter
  • Treatment VI T6 : 30 minutes: Curcumin 2-hydroxypropyl-beta-cyclodextrin. Concen tration 160 micromolar (408 mg complex per liter)
  • Treatment IV was exposed to the treatment solution for 10 minutes, treatment V (T5) was exposed for 20 minutes, and treatment VI (T6) for 30 minutes; after which they were rinsed with seawater, 300ml clean seawater was placed in the blue light treatment. Observations of the number of affected/unaffected lice were conducted at 30, 60, 120, 180, and 240 min, due to lack of visibility counts were made for group 4 and 5 when the treatment solutions were replaced.
  • the riboflavin treatment (T2) had no discernable effect on survival of the salmon lice. All the curcumin cyclodextrin treatments (T4-T6) were effective at reducing survival of the lice, with the majority affected within 60min of exposure to blue light (TablelO). Table 9. Percentage active salmon lice recorded during five observations during a 4h exposure to eight different treatment compounds and three different light sources / conditions
  • curcumin 55.5 mg of compound 3 ([2-hydroxypropyl] beta-cyclodextrin curcumin) and 82.8 mg of compound 7 (beta-cyclodextrin curcumin) were each dissolved in 12 ml. sea water separately using vortex mixer at room temperature. The solutions were saturated solutions. Samples were taken out from the solutions, centrifuged and analysed by HPLC. The peak area for compounds compound 3 and compound 7 was almost 29 and 11 times more than the peak area for curcumin, respectively (including all impurities in all three compounds).
  • Blue light is very effective in degradation of curcumin.
  • the curcumin cy- clodextrin complexes are much more stable than curcumin in presence of blue light.
  • Compound 7 (MH-107) comprising beta-cyclodextrin is much more stable than com pound 3 (MH-103) comprising 2-hydroxypropyl-beta-cyclodextrin in presence of blue light.

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Abstract

La présente invention concerne une composition sous la forme d'une solution aqueuse comprenant un photosensibilisateur destiné à être utilisé dans un procédé de thérapie photodynamique pour une infection par un crustacé parasite externe chez des salmonidés, ladite infection par un crustacé parasite externe comprenant une infection par des poux du saumon, Lepeophtheirus salmonis, Caligus rogercresseyi et Caligus spp. Ledit photosensibilisateur est la curcumine ou un dérivé pharmaceutiquement acceptable de celle-ci, et ledit photosensibilisateur est administré dans un traitement par bain des salmonidés nécessitant un tel traitement, et les poux du saumon sont éclairés par la lumière.
PCT/NO2019/050279 2018-12-17 2019-12-17 Composition destinée à éliminer les crustacés ectoparasites des salmonidés d'élevage à l'aide de la curcumine ou d'un dérivé pharmaceutiquement acceptable de celle-ci et de la lumière WO2020130843A1 (fr)

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CA3123798A CA3123798A1 (fr) 2018-12-17 2019-12-17 Composition destinee a eliminer les crustaces ectoparasites des salmonides d'elevage a l'aide de la curcumine ou d'un derive pharmaceutiquement acceptable de celle-ci et de la lum iere
EP19901125.5A EP3897114A4 (fr) 2018-12-17 2019-12-17 Composition destinée à éliminer les crustacés ectoparasites des salmonidés d'élevage à l'aide de la curcumine ou d'un dérivé pharmaceutiquement acceptable de celle-ci et de la lumière
DKPA202170358A DK202170358A1 (en) 2018-12-17 2021-07-05 Composition for removing crustacean ectoparasites from farmed salmonid fish using curcumin or a pharmaceutical acceptable derivate thereof and light

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WO2021256938A1 (fr) * 2020-06-17 2021-12-23 Mowi ASA Procédé d'élimination de crustacés ectoparasites de salmonidés d'élevage à l'aide de composés de tétrapyrrole hydrosolubles et de lumière
EP4167729A4 (fr) * 2020-06-17 2024-06-26 Mowi ASA Procédé d'élimination de crustacés ectoparasites de salmonidés d'élevage à l'aide de composés de tétrapyrrole hydrosolubles et de lumière
CN113698285A (zh) * 2021-07-28 2021-11-26 浙江海洋大学 一种卤代姜黄素衍生物、其制备方法及其在水产品保鲜上的应用
CN113698285B (zh) * 2021-07-28 2023-02-10 浙江海洋大学 一种卤代姜黄素衍生物、其制备方法及其在水产品保鲜上的应用
CN113845460A (zh) * 2021-07-29 2021-12-28 浙江海洋大学 一种高光敏杀菌活性的姜黄素衍生物、制备方法、鲜虾光动力杀菌保鲜方法
CN113845460B (zh) * 2021-07-29 2023-02-14 浙江海洋大学 一种具有光敏杀菌活性的姜黄素衍生物、制备方法、鲜虾光动力杀菌保鲜方法

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EP3897114A4 (fr) 2022-08-24
NO346510B1 (en) 2022-09-12
EP3897114A1 (fr) 2021-10-27

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