GD INT NATIONAL APPLICATION PUBL D UNDER THE PATENT COOPERATION TREATY (PCT) World Intellectual Property > era Z AAU International Burau Zz (Internationa Publication Number 17 March 2022 (17.03.2022) WIPO! PCT Oz eue! (1) International Patent Classificato ARK 20710 2006.01) A23K 5080.2016.01) 21) International Application Number: PCT/NO2021/050189 (22) International Filing Date: 10 September 2021 (10.09.2021) 25) Filing Language: English (26) Public Langa English 0) Priority Data: 20201004 11 Septem + 2020 (11,09,2020) NO om (72) Inventors: RIKHARDSEN, Bjorn-Inge; Karlsholmveien 6, 8820 Donm (NO), RIKHARDSEN, Berit Anne, sholnveien 6, 8820 Donsia (NO), (74) Agent: BRYN AARFLOT AS; Siomtingsgata 8, 0161 OS- LO WO), Designated States (unless otherwise indicated, for every Kind of national protection available): AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, BZ,, CA.CH, CL, CN, CO, CR, CU; CZ, DE, DJ, DK, DM, DO, DZ, EC, BE, EG, ES, Fl, GB, GD, GE, GH, GM, GT, HIN, HR, HU, ID, IL, IN, IR.IS, IT, J0, JP, KE, KG, KH, KN, KP.KR, KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, MG, Mik, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA.SC, SD, SE, SG, SK, SL, ST, SV, SY, TH, TI, TM, TN, TR.TT-1Z, UA,UG, US, UZ, VC, VN, WS.ZA,ZM.2W. Designated States (unless otherwise indicated. for every Kind of regional protection avaitabley: ARIPO (BW, GH. GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, $2,1Z, UG, ZM, ZW), Enrasian (AM, AZ, BY, KG, KZ, RU. TI. ‘TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, DK, BE, ES, Fl, FR, GB, GR, HR, HU, IE, 1S, IT, LT, LU, LV, MC, MK, MT, NL, NO, PL, PT, RO. RS, SE, SI. SK, SM. TR), OAPI (BF, BI, CF, CG, Ci, CM, GA. GN, GQ, GW, KM, ML, MR, NE, SN, TD, T), ed — with international search report fr, 21(3)) = wit amended claims (Art. 19/1) ;D FOR TREATMENT OF ECTOPARASITE INFECTIONS (7) Abstract: The present invention relates to a medicated fish feed for use in preventing and treating of ectoparasite infections in fish, in parcular of salmon lice infections inaquaculture of Atlantic salmon, The medicated fish feed comprises as active agent ada, seleced from chitin synthesis inhibitors, avermectins and neonicotinoids, Themedicated fish feed is a floating pellet which lus a at least neutal buoyancy and more preferably a positive buoyancy Wo 2022/055360 A1 lll20 30 WO 2022/055360 PCT/NO2021/050189 FISH FEED FOR TREATMENT OF ECTOPARASITE INFECTIONS Field of the invention The present invention relates to a floating feed comprising a chitin synthesis inhibitor, an avermectin, or a neonicotinoid as active ingredient for treatment of ectoparasitic infections in fish, especially of infections of salmonids by the ectoparasitic lice Lepeophtheirus salmonis. Background of the invention For the time being, Atlantic Salmon, Salmo salar, in the on-grown and post- smolt stages are mainly cultivated in open net-pens/cages, which are submerged in the sea, These net-pens are dependent on a free-water exchange with their surroundings through the net to guarantee a sufficient supply with fresh and oxygen- rich water at any time. The water exchange in and out of the cage will be affected by local currents, waves, wind, and tides. Open sea cages have the drawback that they are not isolated from the environment. Thus, small particles and planktonic stages including poisonous microalgae, or the infectious stages of salmon lice, virus, and bacteria can pass with the water through the net meshes, On the other side, excrements, non-eaten feed residues, chemicals, and agents used in farming processes and for treatment of fish and any other small particulate waste may be transported with the water current out of the cage and will be spread to the surroundings. Discharges from aquaculture production are recognized as a major problem due to potential negative impact and effects on the environment such as the water quality, sediments and wildlife in vicinity to the aquaculture site. Parasitic infections in intensive farming of fish such as the Atlantic salmon, Salmo salar, are one of the main problems in salmon aquaculture. In particular infections by ectoparasitic copepods Lepeophtheirus spp. and Caligus spp. Especially the Salmon lice Lepeophtheirus salmonis, as well as the sea lice Caligus elongatus are a major challenge for the industrial production. They are affecting fish welfare and health and as a consequence cause considerable economic losses in the production.20 25 30 Wo 2022/0 PCT/NO2021/050189 2 Salmon lice, Lepeophtheirus salmonis, is an ectoparasite, which belongs to the family Caligidae. Salmon lice have eight different stages in their life cycle with ecdysis in between. Adult salmon lice have eggs strings comprising developing embryos, which after hatching pass through a pelagic phase with two nauplii stages and one copepodite stage. These three stages are free-living planktonic stages. They are passively transported and spread by the water current and cause the spreading of the disease between fish and locations. The copepodite stage, lasting about 10 days at 12°C, is the stage that finally infects the fish by attaching to its skin After attachment, the louse develops further and harms the fish by damaging/feeding on the skin and sucking its blood. Resulting skin lesions can additionally be infected by bacteria causing even more harm to the fish. Lice may also act as vectors for certain bacterial and virus infections such as the ISA-virus (Infectious Salmon Anemia virus) Treatment of infected fish is important to keep control of the pest and there is a strong focus on the development of effective and environmentally sustainable methods for prevention and treatment of the ectoparasite infections. Pest management of lice infections has become a major focus area in salmon farming Salmon lice infections have previously mainly been treated using chemical drugs and agents for delousing, by bath treatment of infected fish. However, in the later years, lice have increasingly developed resistances against most of the commonly known agents used to combat lice infections. Moreover, many of the known agents represent a danger for the environment also affecting the ecosystem in vicinity to aquaculture sites Use of cleaning fish, such as of the wrasse family, feeding on e.g. lice disturbing larger fish, are considered environmentally friendly and have proven to be effective in keeping down the infectious rates with salmon lice, The drawback of such cleaning fish is their availability, high costs in their cultivation as well as a problematic fish welfare of these fish during use and handling. Nowadays, also mechanical and physical methods for removal of lice are increasingly used as an alternative in the treatment of lice such as laser treatment, brushing, treatment with water nozzles, freshwater, warm water ete. A drawback of many of these mechanical and physical methods is the risk to harm the fish and the10 20 Wo 2022/0 PCT/NO2021/050189 caused stress during the treatment and handling as they require a physical interaction with the fish in order to remove the attached lice Effective agents for prevention and treatment of lice infection may also be supplied as orally administered drugs. The least stressful and most effective method is to simply administer a drug comprised in a fish feed such as in form of a medicated feed pellet. Medicated feed pellets comprising an effective agent are thereby distributed in a suitable amount in the cage with the farmed fish to be treated and the feed pellets are consumed by the fish. One problem with the oral administration of drugs in intensive salmon farming are feed losses. Feed pellets not readily consumed by fish sink and may also disintegrate, Feed pellets and smaller particles from the feed pellets may be passively transported out of the cage by currents. Thereby, the active agents will be spread to the surrounding water with all the negative consequences this may have on the food chain, wildlife and ecosystem. Moreover, they may accumulate in sediments and pollute the water over time. An exact control of the amount of ingested medicated feed is very important not only to reduce losses, but also to obtain and ensure the desired therapeutic dose and thus also effect. Loss of medicated feed which sinks makes it very difficult to assess whether the fish have ingested the correct and effective amount of the therapeutic agent. Insufficient therapeutic treatment with an agent is also problematic for the development of reduced sensitivity and resistances of parasites against a drug. Many of the effective agents used today in treatment of salmon lice infections are belonging to the group of chitin synthesis inhibitors (CSIs, also called Chitinase inhibitors) which act by interfering with chitin formation and molting of the copepod. Known CSIs are diflubenzuron and teflubenzuron. Another effective agent used is the avermectin emamectin benzoate. Emamectin benzoate acts by inhibiting muscle contraction by causing a continuous flow of chlorine ions in the GABA and H- Glutamate receptor sites. These are highly effective in the treatment of salmon lice infections. A disadvantage of these agents is that they are unspecific in their mode of action, which means that they not only affect the ectoparasitic lice infested to the fish, but also any other crustacean in case they were to be exposed to the drugs.10 20 Wo 2022/0 PCT/NO2021/050189 Another very effective drug for treatment of salmon lice infection is imidacloprid which is a systemic insecticide belonging to the class of neonicotinoids acting as a neurotoxin. Like the CSIs this agent is not specific for only copepods but will affect other organisms such as other crustaceans if these were to be exposed to the agent, WO2015/198247 discloses a fish feed comprising neonicotinoid, especially imidacloprid, for preventing and treating of parasite infections in fish such as salmon live. Neonicotinoids are added to a premix, which is applied to feed granules or pellets consumed by the fish. The concentration of imidacloprid is preferably about 40 wt%, based on the total weight of the composition. One of the main problems addressed in WO2015/198247 is to provide a suitable mode of oral administration of the drug in fish feed, where the feed is accepted by the fish and consumed/ingested with reduced feed losses. US2015022931 A1 concerns the use of clothianidin for controlling sea lice in a fish population, such as by feeding the fish with a medicated fish feed. EP1168931 B1 concems methods for reducing or preventing parasites in a fish population, feeding emamectin or a salt thereof to the fish population. Neither of the above-mentioned documents address the fact that active agents may be spread to the surrounding water routed through uneaten pellets, with all the negative consequences this may have on the environment and ecosystem. Chemicals such as delousing agents which are toxic to the targeted species may also be toxic to non-target species if they are released into the environment. Studies have shown that there is a risk of negative environmental effects when delousing agents enter the surrounding water. Many studies have addressed effects of neonicotinoids on terrestrial non-target species. There are also data available for aquatic systems, indicating that this is both a problem in freshwater and marine aquatic systems. Many studies have been performed on aquatic crustaceans, in particular in fresh water, which show that imidacloprid induces toxic effects at minimal levels in several aquatic organisms indicating a high sensitivity. This further supports the need for solutions where discharge of this type of unspecific drugs is minimized or excluded as much as possible10 20 Wo 2022/0 PCT/NO2021/050189 Objects of the present invention There is a need to provide improved methods for therapeutic treatments with neonicotinoids, avermectins and CSIs used in treatment of ectoparasitic copepod infection of fish, especially of salmon lice infection of salmon. The present invention aims at providing an environmentally friendly method for salmon lice treatment by oral administration of neonicotinoids, especially imidacloprid, avermectins and/or CSls whereby discharges of active ingredients to the environment are minimised or substantially excluded. The term "treatment" used herein comprises either of prevention or of treating the infection. In particular, the present invention aims at a better control and reduction of feed loss and discharge of drug to the environment when orally administering named drugs to fish. There is a need to improve the control of feed intake when orally administered medicated feed is fed to fish to obtain the desired therapeutic effect. There is also a need to provide improved methods for oral administration of feed in order to reduce the risk for development of resistances and reduced sensitivity. Summary of the invention The present invention solves one or more of the above mention problems. The scope of the present invention is defined by the independent claims. Preferred embodiments are defined in the dependent claims, Thus, the present invention relates in a first aspect to a medicated fish feed for use in preventing and/or treating of ectoparasite infections in fish. The medicated fish feed comprises as active agent a drug selected from chitin synthesis inhibitors, avermectins and neonicotinoids. The fish feed is a floating pellet having at least a neutral buoyancy and more preferably having a positive buoyancy. Preferably, the active agent is selected from imidacloprid, emamectin benzoate, diflubenzuron, and teflubenzuron The floating feed pellet is preferably coated with a composition comprising said active agent and is additionally coated with a fish oil as a second coating. The fish oil serves as an attractant and serves as a barrier for leakage of the active ingredient. Preferably, the medicated feed is used for prevention and treatment of lice infections of salmonids, more preferred for Atlantic salmon Salmo salar.10 20 WO 2022/055360 PCT/NO2021/050189 Preferably, the ectoparasite is a species selected from Lepeophtherius spp. and Caligus spp., (e.g. one of the species Caligus elongatus and Caligus rogercresseyi), more preferably Lepeophtheirus salmonis. A preferred daily dose administered to the fish is 0.5-4 mg per kg fish per day for 7-21 days for imidacloprid; 25-400 ug per kg fish per day for 7-21 days for emamectin benzoate; 1-100 mg per kg fish pr. day for 7-21 days for teflubenzuron; or 1-100 mg per kg fish per day for 7-21 days for diflubenzuron, The medicated feed is particularly useful in the treatment of fish kept in an open net pen. Ina further aspect, the present invention relates to a medicated fish feed comprising as active agent a drug selected from Chitin synthesis inhibitors, avermectins and neonicotinoids, characterized in that the fish feed is a floating pellet with at least a neutral buoyancy and more preferably with positive buoyancy, Preferably, the active agent is selected from imidacloprid, emamectin benzoate, diflubenzuron, and teflubenzuron, most preferably imidacloprid In yet a further aspect, the invention provides a method for producing a medicated floating fish feed, comprising as active agent a drug selected from Chitin synthesis inhibitors, avermectins and neonicotinoids. Using a floating pellet has the great advantage that the pellet is not sinking down when not readily consumed by the fish. Feed pellets commonly used in salmon production, as well as the pellets disclosed in WO2015/198247, have the disadvantage of a negative buoyancy. Uneaten pellets may sink to the bottom or they may be taken by the water current and become unavailable for the fish in the pens. In the best case, unconsumed pellets are stable and will not disintegrate and can eventually be collected together with other waste in the bottom area of the cage. However, on their way down they may also be transported by the water current out of the net cage and may be spread into the water surrounding the cage and environment. If the pellets are not stable, they may also disintegrate into smaller particles and/or the drug may leak out and is thereby spread into the environment, This can have a negative environmental impact on the wildlife where the drug is10 20 Wo 2022/0 PCT/NO2021/050189 spread to. As these drugs are unspecific in their mode of action, they may affect other crustaceans and planktonic species which are likewise sensitive to the drugs. Floating pellets have the advantages that they allow observation of the feeding behavior of the fish, improving the control of feed intake and that correct overall dosage is achieved. The use of floating pellets thus provides an improved control of administered amount of therapeutic agent. This is not only crucial to achieve an effective treatment, but it contributes to avoid reduced sensitivity and resistance over time for a defined drug Unconsumed feed floating at the surface can be easily collected and can be quantified or optionally even refed. Moreover, since medical feed is expensive considerable costs can be saved having a better control and reducing losses. This is thus a new and more effective way and method to administer these unspecific orally administrated drugs to fish in intensive production and solves the above-mentioned problems occurring particularly in open net pens used today. Due to the large volumes and high number of fishes in the cages in today’s intensive production, discharges from medical treatment represent a threat to the ecosystem and environment, Description of the diagrams Embodiments of the present invention will be described below. The following diagrams are referred to in the detailed description, wherein: Figure 1 shows time series of the maximum concentration of imidacloprid per kilogram sediments over time when including feed waste of 2 % (sinking feed pellets); Figure 2 shows time series of the maximum concentration of imidacloprid per kilogram sediments over time when excluding feed waste (floating feed pellets). Detailed description of the invention and of preferred embodiments The present invention will be further explained using the ectoparasitic copepod salmon lice Lepeophtheirus salmonis and the Atlantic salmon as an example. However, the skilled person will understand that the described medical feed, use and methods of treatment can also be used for other salmonids or fish10 20 Wo 2022/0 PCT/NO2021/050189 species infected with these ectoparasites or other parasites which can be treated by medicated feed according to the invention. The invention will, however, be limited to the use in species which ingest feed at the water surface/upper water layers since feed particles according to the invention are floating. Preferred ectoparasite infections to be treated by the medicated feed according to the present invention are ectoparasites belonging to Lepeophtheirus spp. and Caligus spp Preferred drugs comprised in the medicated feed are all belonging to the group of chitin synthesis inhibitors (CSIs), avermectins or neonicotinoids (i.e imidacloprid). Preferred active ingredients for treating infections according to the present invention are imidacloprid, emamectin benzoate and di- and teflubenzurone Emamectin benzoate is an avermectin, while di- and teflubenzurone are belonging to the group of benzoylurea. Particularly preferred is imidacloprid as active agent in the disclosed medicated feed according to the invention Features and production of the medicated feed There are different methods for production of a medicated feed that meet the properties of the feed according to the present invention. In common for all methods and an essential feature of the present invention, is that the feed pellet comprising the drug for treating ectoparasite infections is a floating pellet, i.e. a feed pellet or particle which has at least a neutral buoyancy and more preferably a positive buoyancy, Production of floating feed and medicated feed Use of floating feed for feeding fish or aquatic organisms is known, particularly for allowing observation of the feeding behavior of the fish, improving the control of feed intake. Different methods to produce feed particles with a neutral or positive buoyancy exist. Floating characteristics of feed pellets may be adjusted by regulation of the expansion rate of the pellet during its production. The expansion rate of the pellet both depends on ingredients used in the feed formulation, in particular the amount and source of starch, as well as the temperature or the steam pressure in front of the extruder die, Bulk density is a crucial parameter for controlling the10 20 WO 2022/055360 PCT/NO2021/050189 buoyancy, i.e. defining whether the pellets will be floating or sinking. High temperature in the process result in more expansion, a low bulk weight and a floating pellet. Typical standard methods used to produce floating fish feed pellets are described in teaching books in the field and are thus well known to the skilled person in the field. Generally, a feed mash formulation of dry (without or with liquid ingredients) can be extruded into a highly expanded floating aquatic feed (320-500 gil), in accordance with the feed of the invention, or a sinking pellet (550-700 g/l), by adjusting temperature and steam pressure during the extrusion process. One description of the procedure for production of highly expanded floating aquatic feeds describe that it requires 200-250 square millimeters of die open area per metric ton of throughput. Expansion of the extrudate after it passes through the die results in a product with a bulk density of 320-400 grams per liter and 21-24 % moisture (Feed manufacturing Technology IV, 4" edition, American Feed Industry Association 1994 manufacturing of floating pellets disclosed on p. 514). An example, how a suitable floating feed can be produced is also disclosed in a recent publication by Welker et al. 2018 (Welker, T.L., Overturf, K., Snyder, S., Liu, K., Abemathy, J., Frost, J., Barrows, F.T., 2018. Effects of feed processing methods (extrusion and expansion- compression pelleting) on water quality and growth of rainbow trout in a commercial setting. Journal of Applied Aquaculture), The described method for the production of floating and sinking feed uses an extruder with five-barrel sections. The floating feed was produced at 119°C with the last section heated up to 131°C. The retention time was 18 s and screw speed was 544 rpm. The same receipt was produced into a sinking feed by processing the mash at 116°C in the barrel but cooling the last section to 17°C. The retention time in the extruder was 18 s and the screw speed was reduced to 509 rpm More particularly, as stated above, bulk density is crucial for controlling the buoyancy of the pellet. Fish feed pellets typically have a bulk density ranging between 200 — 700 gil. A lower bulk density indicates a highly expanded porous product that floats, while high bulk density indicates compact feed pellets with less pores. A bulk density lower than about 450 g/l will have positive buoyancy. In one10 Wo 2022/0 PCT/NO2021/050189 10 embodiment, the feed pellet of the invention has a bulk density of 200-500 gil, such as 320-500 gil, and more preferably of 320-450 g/l Bulk density is controlled by expansion rate of the pellet, i.e. the diameter of the pellet to diameter of the extruder die opening. The expansion rate of high energy feed which is coated with oil post extrusion in a vacuum coater, is normally ranging between 15- 25%. The more expanded pellet is more likely to have a positive buoyancy. The expansion rate is controlled by several parameters in the feed production, including the following a) Amount of starch in the formul n, i.e. in the basal feed material, herein called carrier material, A formulation with a starch inclusion > 12% of the dry matter basis is more likely to have positive buoyancy. Formulations with the purpose of producing floating pellets often contain about 20% starch depending on the species. Carnivore fish like Atlantic salmon and rainbow trout should however have lower levels of starch in the formulation. In one embodiment, the formulation of the feed pellet of the invention comprises 12-20% starch, such as 12 - 16 % starch 'b) Starch source — starch is added to feed for carnivore fish as a functional ingredient to ensure proper binding and expansion. The combination of heat and water in the extruder system will result in swelling of starch granules, leaking of amylose and amylopectin until the granular structure is lost. Amylose and amylopectin are important for the viscosity of the dough in the extruder. Starch sources with large starch granule create higher viscosity and have potential to support greater expansion. Starch of different origin is used in different fish feed products. The starch and protein matrix of the formulation is important for the expansion rate and final structure of the feed. In one embodiment, the feed pellet of the invention comprises starch selected from amylose and amylopectin. — c) Expansion is also controlled by energy generated in the extruder. Energy originates from two sources in expanded fish feed - thermal energy and mechanical energy. Thermal energy can be injected in the pre-conditioner as steam or injected directly into the extruder barrel or via heating of the jackets enclosing the screw(s). Mechanical energy is generated by the rotating screws. The specific mechanical energy generated in the extruder is determined by the configuration of the extruder (e.g. screw configuration), the formulation, and adjustable parameters in10 20 WO 2022/055360 PCT/NO2021/050189 1 the extrusion process for example screw rotation speed, moisture and temperature. The floating fish feed of the invention may typically be produced with temperature in front of the extruder die > 120°C, and a pressure in front of the die > 20 bar. The fish feed has otherwise standard compositions known in the art and will be adapted to be suitable for feeding of salmon or any other fish species to be treated. The same applies to the chosen size of the pellet which will be adjusted to. be suitable for oral administration to fish There are different methods for how the medication (API/drug) according to the present invention can be added to the floating feed pellet (also called carrier). In a preferred method the drug can be added to a feed carrier which is a defined fish feed used to produce the medicated feed pellet. The pellet will thereafter be coated by a fish oil. Typical steps are: The feed pelleticarrier is controlled, weighed, and added to a mixer. The active pharmaceutical ingredient (APUdrug) is controlled, weighed, added and mixed together with the medicated feed carrier in the mixer. Final step is the spraying (coating) of fish oil through nozzles in the mixer. Suitable mixing times are defined for each step. Before the batch leaves the mixer, samples for analyzes are taken out by a separate sample collector. Each batch has to be analyzed for its API content confirming that the medicated feed is within the specifications. The invention hence provides a method for producing a fish feed pellet, comprising as active agent a drug selected from chitin synthesis inhibitors, avermectins, and neonicotinoids, and having at least neutral buoyancy and more preferably a positive buoyancy, comprising the steps of; a) extruding a dry feed mash formulation into highly expanded pellets; b) adding the drug selected from chitin synthesis inhibitors, avermectins, or neonicotinoids to the pellets of step a); c) optionally coating the pellets of step b) with an oil, preferably a fish oil Vacuum coating of feed pellets Vacuum coating is a standard process in the production of high energy aquatic feeds and is also used in the production of medicated feeds. This is a10 20 Wo 2022/0 PCT/NO2021/050189 12 method which is well known to the skilled person. Heat sensitive nutrients can also be applied in post extrusion coating operations. Extruders are typically used to produce the pellets (carrier) in order to get the right physical properties of an expanded basal feed pellet. The expanded pellets contain pores which can be filled with oil during a vacuum coating process. The dried pellet, containing about 8-10% moisture is transferred to the vacuum coater. The vacuum coater is an air-tight mixer e.g. ribbon or paddle type mixer. Rotating drums or devices inside the drum, will make sure that all pellet surfaces are exposed to a mist of oil sprayed into the drum through nozzles. A negative air pressure is generated inside the drum using a vacuum pump removing the air. When vacuum is gradually applied to the airtight closed drum, the pressure drops and the air bubbles are leaving the pellets, ascending through the layer of oil to the top. The constant mixing of pellets distributes the oil on all pellet surfaces. By slowly opening the air inlet the negative pressure will gradually disappear until the pressure in the drum equals to the surrounding air pressure. Oil is gradually sucked into the pellet as vacuum is released The vacuum coating principle can also be used for application of additives or to produce medicated feed, Different components can be applied in layers to the pellet by use of double coating, The final product, i.e. the feed pellet comprising both as active agent a drug selected from chitin synthesis inhibitors, avermectins and neonicotinoids, and any coating of fish oil, has a neutral or positive buoyancy. Hence, appropriate measurements are to be taken during the process of applying the active agent and any fish oil, ensuring that the bulk density of the final feed pellet is such that the pellet will float, i.e. that the bulk density of the feed pellet is in the range of 200-500 gil In one embodiment, the invention hence comprises a method for producing a fish feed pellet, in accordance with the description above, comprising as active agent a drug selected from chitin synthesis inhibitors, avermectins, and neonicotinoids, and having a at least neutral or positive buoyancy, comprising the steps of; a) extruding a dry feed mash formulation into highly expanded pellets;1s 20 25 30 35 WO 2022/055360 PCT/NO2021/050189 13 b) vacuum coating the pellets of step a) with a drug selected from chitin synthesis inhibitors, avermectins, and neonicotinoids; c) optionally vacuum coating the pellets of step b) with an oil, preferably a fish oil, Preferably, the produced feed pellets have a positive buoyancy. Preferred dosage regimes (daily dose) comprised in the medical feed according to the invention imidacloprid Dosage of 0.5-4 mg per kg fish per day for 7-21 days. Emamectin benzoate: Dosage 25-400 ug per kg fish per day for 7-21 days. Teflubenzuron: Dosage 1-100 mg per kg fish pr. day for 7-21 days Diflubenzuron’ Dosage 1-100 mg per kg fish per day for 7-21 days. The above given dosages are given per kg body wet weight of fish Feeding to the fish in a fish cage The floating pellets can be distributed to the fish by suitable known feeding distributors either at the water surface or under the water surface. Unconsumed pellets will float up. If not consumed over longer periods, pellets can be collected and quantified. The medicated feed can be fed out continuously or batchwise. Preferably the feed is supplied continuously to the fish during the whole day Experimental section In an Environmental Risk Assessment (ERA) for imidacloprid, it was simulated a scenario where 2% of the medicated non-floating feed with imidacloprid as active agent sank down to the sediment as feed loss and a scenario where medicated floating feed pellets according to the present invention with imidacloprid as active agent were used without any feed losses. The feed was administered for 7 days10 WO 2022/055360 PCT/NO2021/050189 14 (March 2 to March 9) and the concentrations in the sediments under the net cage was assessed until April 20. Even with this low amount of feed loss of only 2 %, differences in the accumulation rate of imidacloprid in the sediments were found over time for the different feed types (see figure 1 for scenario with 2 % feed loss and figure 2 without feed loss). Since feed losses in open-net aquaculture are typically about 10-15 % for sinking feed pellets, the accumulation of the agent in sediments will in reality be much higher as well as the potential negative impact on the ecosystem10 20 WO 2022/055360 PCT/NO2021/050189 15 Claims 1. A medicated fish feed for use in preventing or treating of ectoparasite infections in fish, where the medicated fish feed comprises as active agent a drug selected from chitin synthesis inhibitors, avermectins, and neonicotinoids, wherein the fish feed is a pellet with a at least neutral buoyancy and more preferably a positive buoyancy. 2. A medicated fish feed according to claim 1, for use according to claim 1, wherein the active agent is selected from imidacloprid, emamectin benzoate, diflubenzuron, and teflubenzuron. 3. Amedicated fish feed according to claim 1 or 2, for use according to claim 1, wherein the feed pellet is coated with a composition comprising said active agent and is additionally coated with a fish oil as a second coating 4. Amedicated fish feed according to any of the preceding claims for use in prevention and treatment of lice infections of salmonids. 5. Amedicated fish feed according to any of the preceding claims, for use according to claim 1, where the fish is an Atlantic salmon Salmo salar. 6. Amedicated fish feed according to any of the preceding claims, for use according to claim 1, where the ectoparasite is a species selected from Lepeophtheirus spp. and Caligus spp., preferably Lepeophtheirus salmonis. 7. Amedicated fish feed according to claim 2 wherein the daily dose administered to the fish is 0.5-4 mg per kg fish per day for 7-21 days for imidacloprid; 25-400 ug per kg fish per day for 7-21 days for emamectin benzoate; 1-100 mg per kg fish pr. day for 7-21 days for teflubenzuron; or 1-100 mg per kg fish per day for 7-21 days for diflubenzuron,10 20 WO 2022/055360 PCT/NO2021/050189 16 8. Amedicated fish feed according to any of the preceding claims for use in the treatment of fish in a net pen. 9. Amedicated fish feed according to any of the preceding claims, for use according to claim 1, wherein the feed pellet has a bulk density of 320-500 gil 10. A medicated fish feed comprising as active agent a drug selected from Chitin synthesis inhibitors, avermectins and neonicotinoids, characterized in that the fish feed is a floating pellet with a at least neutral buoyancy and more preferably a positive buoyancy. 11. A medicated fish feed according to claim 10, wherein the active agent is selected from imidacloprid, emamectin benzoate, diflubenzuron and teflubenzuron, most preferably imidacloprid 12. Amedicated fish feed according to claim 10 or 11, wherein the feed pellet has a bulk density of 320-500 g/l 13. A method for producing fish feed pellets, the pellets comprising as active agent a drug selected from chitin synthesis inhibitors, avermectins, and neonicotinoids, and having a at least neutral buoyancy and more preferably a positive buoyancy, the method comprising the steps of; a) extruding a dry feed mash formulation into highly expanded pellets; b) adding the drug to the pellets of step a); c) optionally coating the pellets of step b) with an oil, preferably a fish oil 14. A method for producing fish feed pellets according to claim 13, wherein step b) comprises vacuum coating the pellets of step a) with the drug; and the optional step c) comprises vacuum coating the pellets of step b) with an oil, preferably a fish oilWO 2022/055360 PCT/NO2021/050189 "7 AMENDED CLAIMS received by the International Bureau on 14 Jan 2022 (14.012022) 1. Amedicated fish feed for use in preventing or treating of ectoparasite infections in fish, where the medicated fish feed comprises as active agent a drug selected from chitin synthesis inhibitors, avermectins, and neonicotinoids, wherein the fish feed is a pellet with a at least neutral buoyancy and more preferably a positive buoyancy, wherein the feed pellet has a bulk density of 320-500 g/l 2 A medicated fish feed according to claim 1, for use according to claim 1, wherein the active agent is selected from imidacloprid, emamectin benzoate, diflubenzuron, and teflubenzuron. 3. Amedicated fish feed according to claim 1 or 2, for use according to claim 1, wherein the feed pellet is coated with a composition comprising said active agent and is additionally coated with a fish oll as a second coating 4. Amedicated fish feed according to any of the preceding claims for use in prevention and treatment of lice infections of salmonids. 5. Amedicated fish feed according to any of the preceding claims, for use according to claim 1, where the fish is an Atlantic salmon Salmo salar. 6. Amedicated fish feed according to any of the preceding claims, for use according to claim 1, where the ectoparasite is a species selected from Lepeophtheirus spp. and Caligus spp., preferably Lepeophtheirus salmonis. 7. Amedicated fish feed according to claim 2 wherein the daily dose administered to the fish is either of 0.5-4 mg per kg fish per day for 7-21 days for imidacloprid; 25-400 tig per kg fish per day for 7-21 days for emamectin benzoate; 1-100 mg per kg fish pr. day for 7-21 days for teflubenzuron; or 4-100 mg per kg fish per day for 7-21 days for diflubenzuron, AMENDED SHEET (ARTICLE 19)WO 2022/055360 PCT/NO2021/050189 18 8. Amedicated fish feed according to any of the preceding claims for use in the treatment of fish in a net pen. 9. — Amedicated fish feed comprising as active agent a drug selected from Chitin synthesis inhibitors, avermectins and neonicotinoids, characterized in that the fish feed is a floating pellet with a at least neutral buoyancy and more preferably a positive buoyancy, wherein the feed pellet has a bulk density of 320-500 g/l 10. Amedicated fish feed according to claim 9, wherein the active agent is selected from imidacloprid, emamectin benzoate, diflubenzuron and teflubenzuron, most preferably imidacloprid. 11 Amethod for producing fish feed pellets, the pellets comprising as active agent a drug selected from chitin synthesis inhi rs, avermectins, and neonicotinoids, and having a at least neutral buoyancy and more preferably a positive buoyancy, the method comprising the steps of; a) extruding a dry feed mash formulation into highly expanded pellets; b) adding the drug to the pellets of step a); c) optionally coating the pellets of step b) with an oil, preferably a fish oil 12. Amethod for producing fish feed pellets according to claim 11, wherein step b) comprises vacuum coating the pellets of step a) with the drug; and the optional step c) comprises vacuum coating the pellets of step b) with an oil, preferably a fish oil. AMENDED SHEET (ARTICLE 19)WO 2022/055360 PCT/NO2021/050189 WW 8 é 2 S 8 ‘Concentration (gig! 00 imidacloprid per kg sediment OM0F OBE (0829 GAD O4NE Oda O420 Date Fig.1 0903 991G 022 GABD GEOG AND 4zd Daw Fig. 2CORRECTED VERSION NATIONAL SEARCH REPORT Intemational application No, PCT/NO2021/050189 CLASSIFICATION OF SUBJECT MATTER A23K 20/10 (2006.01); A23K 50/80 (2016.01) According to Intemational Patent Classification (IPC) orto both national classification and [PC B. FIELDS SEARCHED “Minimum documentation searched (Classification system Tollowed by classification symbols) A23K 20/10 A2BK S080 ‘Documentation searched other than minimum documentation fo the extent that sch documents are included in the fekis searched DK.NO, SE, Fl Electronic dala base coosulied ring the international search (oame of data base and, where practicable, search terms used) EPODOC, WPI, FULL TEXT, CAPLUS, POSCITECH: ENGLISH, DOCUMENTS CONSIDERED TO BE RELEVANT. Category® Citation of document, with indication, where appropriate ofthe relevant passages Relevant to claim No, WO 2015198247 Al (OALLORAN JOHN [CA]; DROST JOHN TERENCE [CA]) 30 December 2015 (2015-12-30) y ‘Table 1: paragraph [0011-00015], [0064-0067], [0069-0073]. (0028), [0053], (0006- 9.12 (0007: example 1; claim 9, x 1-8, 10-11, 1314 ‘WO 2014064184 Al (NOVARTIS AG [CHI; BOUVIER JACQUES [CH]; NANCHEN STEVE [CH]: PERRET JEAN-LUC [CH] 01 May 2014 (2014-05-01) x Example I: table I; and page 5 second paragraph, 1-8, 10-11, 1314 Y 9.12 WO 2008039172 Al (SZOKE ANNAMARIA [HU}) 13 May 2004 (2008-05-13) x Example 1 and 2: paragraph [00042-0042]: claim 3-7 and 13 1-8, 10-11, 13-14 Y 92 EP 3132684 Al (NOVARTIS TIERGESUNDHEIT AG [CI ‘TIERGESUNDHEIT AG (CH) 22 February 2017 (2017-0 Y Example 1 and : paragraph [00042-0042]; claim 3-7 and 13, 9.12 x 1-8, 10-11, 13-14 7] Further documents ae listed in the continuation of Box C. [J See patent family annex. 7 Special categories of eked dooumes “Tle document published after the intrtional line date or pony de an notin confit wih te pplication bated to undead the +A” document defining tho general sate ofthe at which snot considered Pancple or hey undoing the fveation tobe of particular relevance > dame sty eosin einen apton 3° Ua el lr: he dame ein cao "att plc rt bt puts on or aftr 1 Seatac cake on document of particular relevance; the claimod invention cannot he fensidered te involve a iveative step whea the docuneat is Sombined wth one ce more oer such documents, sich combaation feng covious os person skied inte re document member of the same pata amily 1L° document which may tzow doubts on priority limi) or whichis “Y" Sed To etalinh te publication date of anther tation or ots Special reason as spectiod “0 document refering to a9 oral dielosre, use, exibition or oer mew © 1p" document publsie prior tothe international ing date but ater thn the pron dat lated Date ofthe actual completion of the international search 01 December 2021 Date of mailing of the international search report 01 December 2021 ‘Name and mailing address ofthe ISAIKNN Nordic Patent Institute Helgeshoj AIlé 81, 2630 Taastrup Denmark Telephone No, +45 43 50 85 00 Facsimile No, +4543508008, Fon PCT/ISA/210 (second sheet) uly 2019) “Authorized officer Shire Elmi Telephone No. +45 43 50 85 00INTERNATIONAL SEARCH REPORT Tnierational application No. PCT/NO2021/0S0189 DOCUMENTS CONSIDEI Category® Citation of document, with indication, where appropriate, ofthe relevant passages Relevant to elim No, THE USE OF LUFENURON TO TREAT FISH LICE (ARGULUS SP) IN KOI (CYPRINUS |CARPIO); MAYER , JORG et al. Journal of Exotic Pet Medicine 22 (2013), pp 65-68. 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AL 10 August 2005 cn 172956 A 18 Fanuary 2006 CN toners 22 October 2008, EP 3132684 Al February 2017 WO 2013167640 Al 14 November 2013 DK 201300268 A (9 November 2013 No 20130862 A 28 June 2013 AU 2013258014 AL (09 October 2014 cA 2868038 AL 14 November 2013 cn loi2s4s86 A. 14 Famuary 2015 EP 2846640 AL 18 March 2015 Us 2015125509 Al 07 May 2015, vw 41465 A 25 March 2015, AU 20132580148 B2 (06 August 2015 IP 2015525203 A (08 September 2015, RU 2014148977 27 hune 2016 EP 2846640 BL 19 October 2016 DK 790728 B (09 October 2017 cA 2868938 C 14 November 2017 CN 1845868 28 November 2017 IP 62671888 B2 24 January 2018, 2642637 C2 25 January 2018 2019380981 AL 19 December cA 3066449 AL 03 July 2021 EP 0391213. Al 1B Apel 1994) 5128153 (OT uly 1992 29114 AL 12 November 1992 cA 2072320 A 26 December 1993 DE «6LI38ITE_ 16 November 1995 cA 2072320 17 September 1996 cn 106578708 A 26 Apeil2017 NONE Form PCTASA/210 (patent Family annex) Gly 2019)