Note : Les descriptions sont présentées dans la langue officielle dans laquelle elles ont été soumises.
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USE OF ISOXAZOLINE FOR PROTECTION AGAINST PARASITIC PESTS IN
FISH
RELATED APPLICATION
[0001] This disclosure claims benefit of and priority to
United States
Provisional Patent Application serial no. 63/182,576 filed April 30, 2021,
incorporated herein by reference in its entirety.
FIELD
[0002] The present disclosure relates to methods of use of
compounds for
treating or preventing parasite infection, such as sea lice infection, in
fish, such as
salmon.
BACKGROUND
[0003] Parasites such as sea lice are ectoparasites that
latch onto the skin
of fish during free-swimming larval stages, feeding on fish's blood, mucus,
and
skin, larger numbers of sea lice result in loss of growth performance,
decrease
animal welfare and can cause fish mortality. Ectoparasite infestation in
aquaculture is a significant commercial concern, and an infestation in, for
example,
farmed salmon can affect wild salmon stocks. Economically important species of
sea lice include Lepeophtheirus salmonis and various Caligus species including
Caligus clemensi, Caligus elongatus and Caligus rogercresseyi, can cause
deadly
infestations of wild fish as well as farm-grown fish. Affected fish include
salmonid
such as Atlantic salmon (Salmo salar), Chinook salmon (Oncorhynchus
tshawytscha), coho salmon (Oncorhynchus kisutch), chum salmon
(Oncorhynchus keta), pink salmon (Oncorhynchus gorbuscha), Sockeye salmon
(Oncorhynchus nerka), and rainbow trout (Oncorhynchus mykiss; the anadromous
form of which is known as steelhead trout).
[0004] The control of sea lice in commercial salmon farming
can include
chemical treatments that are applied in a bath by temporarily placing a tarp
around
a cage containing the fish and water or pumping fish into a well boat,
however,
such treatments are restrained by concerns about even mixing of the chemical,
releasing the chemical into the environment and the sea lice developing
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resistance. In particular, where high doses of chemical are employed,
resistance
development could accelerate.
[0005] Fluralaner is an isoxazoline that is generally known
for treating fleas
and ticks in cats and dogs, and for treating mites in poultry, sold under the
tradenames BravectoTM and ExzOltTM. In dogs, medication is given once at 25 to
50 mg/kg of dog weight and it provides protection against flea and tick for 12
weeks. In poultry, the drug is provided in drinking water for one day at 0.5
mg/kg
of chicken weight and the treatment is repeated 7 days later. Current uses of
fluralaner are believed to be limited to domesticated or industrial animals
and
birds, and on crops to treat insect pests.
SUMMARY OF THE DISCLOSURE
[0006] The present inventor has discovered that isoxazoline
such as fluralaner
is an effective treatment for controlling sea lice infestation in fish such as
salmon,
showing high sea lice removal and low wastage of feed.
[0007] Accordingly, the present disclosure includes a use of
an isoxazoline
such as fluralaner, a composition comprising an isoxazoline such as fluralaner
and
at least one excipient, or a medicated fish feed comprising fish feed granules
or
pellets coated or mixed with a composition comprising an isoxazoline and at
least
one excipient, for preventing or treating parasite infection in a plurality of
fish,
wherein the isoxazoline, composition or medicated fish feed is for use at a
daily
dose comprising at least about 0.005 mg isoxazoline (e.g. fluralaner)/kg of
fish, up
to about 5 mg isoxazoline (e.g. fluralaner)/kg of fish, for 5 to 15 days.
[0008] In an embodiment, the isoxazoline comprises
fluralaner, afoxolaner,
lotilaner, or a veterinary acceptable salt of any of the foregoing_ In an
embodiment,
the isoxazoline is fluralaner.
[0009] In an embodiment, the at least one excipient
comprises carrier, pork
liver flavour, sucrose, maize starch, sodium lauryl sulfate, disodium embonate
monohydrate, magnesium stearate, aspartame, glycerol, soya-bean oil,
polyethylene glycol such as Macrogol 3350, vegetable oil, or an approved
pharmacopeia excipient. In an embodiment, the excipient is an approved
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pharmacopeia excipient. In an embodiment, the carrier comprises fish oil, fish
meal, shrimp meal, krill meal, krill oil, poultry meal, protein concentrate,
select
amino acids, water or combinations thereof. In an embodiment, the carrier
comprises a fish oil, optionally further comprises a krill meal.
[0010] In an embodiment, the isoxazoline (e.g. fluralaner),
composition, or
medicated fish feed is for oral use by feeding a divided dose in a plurality
of feeds 1,
2, 3, 4, or 5 times daily, for 5 to 15 days. In an embodiment, the daily dose
comprises
0.01 mg isoxazoline (e.g. fluralaner)/kg of fish, up to about 1 mg isoxazoline
(e.g.
fluralaner)/kg of fish. In an embodiment, the daily dose comprises 0.07 mg
isoxazoline
(e.g. fluralaner)/kg of fish, up to about 0.7 mg isoxazoline (e.g.
fluralaner)/kg of fish.
In an embodiment, the daily dose comprises 0.01, 0.1, 0.5, 0.75, or 1 mg
isoxazoline
(e.g. fluralaner)/kg of fish. In an embodiment, the daily dose comprises 0.1
mg
isoxazoline (e_g_ fluralaner)/kg of fish. In an embodiment, the isoxazoline
(e.g.
fluralaner), composition, or medicated fish feed is for use at a total dose of
about
0.025 mg isoxazoline (e.g. fluralaner)/kg of fish, up to about 75 mg
isoxazoline (e.g.
fluralaner)/kg of fish.
[0011] In an embodiment, the fish is salmonid. In an
embodiment, the
salmonid is a salmon or trout. In an embodiment, the salmonid is Salmo salar,
Oncorhynchus tshawytscha, Oncorhynchus kisutch, Oncorhynchus keta,
Oncorhynchus gorbuscha, Oncorhynchus nerka, or Oncorhynchus mykiss. In an
embodiment, each salmonid has an average mass of less than or equal to 5 kg,
optionally an average mass of from 25 g to 5 kg.
[0012] In an embodiment, the parasite is a sea louse. In an
embodiment,
the sea louse is at any stage of life-cycle. In an embodiment, the sea louse
is pre-
adult sea lice, copepodites, chalimus, adult sea lice, or gravid female lice.
In an
embodiment, the sea louse is Lepeophtheirus or Caligus, optionally the
Lepeophtheirus is Lepeophtheirus salmonis, optionally the Caligus is Caligus
clemensi, Caligus elongatus, or Caligus rogercresseyi.
[0013] Other features and advantages of the present
disclosure will
become apparent from the following detailed description. It should be
understood,
however, that the detailed description and the specific Examples while
indicating
preferred embodiments of the disclosure are given by way of illustration only,
since
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various changes and modifications within the spirit and scope of the
disclosure will
become apparent to those skilled in the art from this detailed description.
[0014]
The present disclosure also includes a fish feed composition (i.e. a
fish feed or a fish feed additive) comprising fluralaner in a dosage described
in this
disclosure. The present disclosure also includes a use of fluralaner for
preventing
or treating sea lice in fish, wherein the fluralaner is administered in a
composition
described in this disclosure. The present disclosure also includes a kit for
preparing a medicated fish feed for reducing or treating parasites in a fish
population, comprising a supply of fluralaner or a salt thereof in a
container, and
printed instructions for feeding the fluralaner at a daily dose per kg of fish
biomass
per day for a total number of days of dosage period as described herein. The
treated fish is optionally a salmon, such as an Atlantic salmon or Pacific
salmon,
or rainbow trout (Oncorhynchus mykiss). It will be appreciated by a person
skilled
in the art that any fish in need of prevention or treatment of infection could
receive
the compositions described herein, including the list of fish in US 5,504,081
columns 5 and 6, and for prevention or treatment of a wide variety of
parasites
including the genera of parasites listed in column 5 of US 5,504,081, and the
foregoing is incorporated herein by reference.
DETAILED DESCRIPTION
[0015]
The disclosure relates to compositions and methods of using the
isoxazoline class of insecticides in fish feed to control parasites, typically
saltwater
crustacean parasites. Optionally, certain freshwater parasites may also be
treated.
This class of products includes, but is not limited to, the compounds
fluralaner,
afoxolaner, and lotilaner. These insecticides control the pest, such as sea
lice, by
impairing the nervous system of insects and crustaceans.
I. Definitions
[0016]
Unless otherwise indicated, the definitions and embodiments
described in this and other sections are intended to be applicable to all
embodiments and aspects of the present disclosure herein described for which
they are suitable as would be understood by a person skilled in the art.
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[0017] Terms of degree such as "substantially", "about" and
"approximately"
as used herein mean a reasonable amount of deviation of the modified term such
that the end result is not significantly changed. These terms of degree should
be
construed as including a deviation of at least 5% of the modified term if
this
deviation would not negate the meaning of the word it modifies.
[0018] As used in this disclosure, the singular forms "a",
"an" and "the"
include plural references unless the content clearly dictates otherwise.
[0019] The recitation of numerical ranges by endpoints
herein includes all
numbers and fractions subsumed within that range (e.g. 1 to 5 includes for
example 1, 1.5, 2, 2.75, 3, 3.90, 4, and 5). It is also to be understood that
all
numbers and fractions thereof are presumed to be modified by the term "about".
[0020] The term "coat" or a derivative thereof as used
herein as related to
fish feed granules or pellets refers the granules or pellets being covered or
having
spread over their outer surface, for example, an active ingredient or
composition
of the present disclosure. Such covering or spreading includes spraying or
dusting the granules or pellets with the active ingredient or composition (or
rolling
the granules or pellets in the active ingredient or composition), or for
sealing the
active ingredient or composition onto the feed with applications of edible
fats. An
alternative to coating is to mix the active ingredient or composition with
feed
ingredients that are then extruded into feed granules or pellets.
[0021] The term "excipient" or a derivative thereof as used
herein refers to
a substance formulated with the active ingredient of the present disclosure.
An
excipient can be, but is not limited to, anti-adherents, coatings, colours,
disintegrants, flavors, glidants, lubricants, preservatives, sorbents,
sweeteners,
attractant, or carrier. For example, an excipient can be pork liver flavour,
sucrose,
maize starch, sodium lauryl sulfate, disodium embonate monohydrate,
magnesium stearate, aspartame, glycerol, soya-bean oil, polyethylene glycol
such as Macrogol 3350, vegetable oil, or an approved pharmacopeia excipient.
Pharmacopeia excipient promotes stability and uniform mixing with the active
ingredient_ An approved pharmacopeia excipient can be an approved excipient
from US Pharmacopoeia (USP). For example, an approved pharmacopeia
excipient of the present disclosure includes a non-GMO dextrose such as
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Emdex . Excipients such as carriers in a composition or medicated fish feed
can
help consumption of the feed in fish. Such carriers include fish oil, fish
meal,
shrimp meal, krill meal, krill oil, poultry meal, protein concentrate, select
amino
acids, and/or water. The protein concentrate can be processed protein
concentrate such as processed corn protein concentrate or processed soybean
protein concentrate, for example, at 80%, 85%, 90%, 95% or 99% protein, on a
dry weight basis, and/or with less than 1% starch on a dry weight basis. For
example, the carrier can be processed corn protein concentrate having at least
75.0% crude protein, at least 2.0% fat, at most 2.5% fiber and at most 1.0%
starch, on a dry weight basis. In an embodiment, the carrier comprises fish
oil,
fish meal, shrimp meal, krill meal, krill oil, poultry meal, protein
concentrate, select
amino acids, water or combinations thereof. In an embodiment, the carrier
comprises a fish oil, optionally further comprises a krill meal.
[0022] References to a kg of fish should be understood as
references to a
kg of fish biomass. References to "fish" should be understood as references to
a
population of fish, except where the context indicates that a single fish is
intended.
II. Methods and Uses
[0023] The present inventors disclose that using or
administering
isoxazoline such as fluralaner is effective in preventing or treating parasite
infestations (e.g. sea lice infections) in fish. The isoxazoline can be fed to
the fish
in pure form, in a composition comprising at least one excipient, or in a
medicated
fish feed that has fish feed coated or mixed with the isoxazoline-containing
composition. For example, feeding isoxazoline such as fluralaner to fish in
pure
form or in a form of a suitable medicated fish feed decreases parasite
infestations
and maintains good food uptake. The term "pure" does not require absolute
purity, but rather it is intended as a relative definition, and can include
isoxazoline
that is highly purified or isoxazoline that are only partially purified, as
those terms
are understood by those of skill in the relevant art and within acceptable
level of
tolerance, for example, when it contains no other chemicals that might
interfere
with its action. For example, a claimed isoxazoline which has a purity of
100%,
or at least 99.999%, 99.99%, 99.9%, 99%, 95%, or 90% by weight or greater is
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contemplated. Accordingly, the present disclosure includes a method of
preventing or treating parasite infection in a plurality of fish, comprising
administering to the fish an effective amount of an isoxazoline such as
fluralaner,
a composition comprising an isoxazoline such as fluralaner and at least one
excipient, or a medicated fish feed comprising fish feed granules or pellets
coated
or mixed with a composition comprising an isoxazoline such as fluralaner and
at
least one excipient, wherein the isoxazoline, composition, or medicated fish
feed
is administered at a daily dose comprising at least about 0.005 mg isoxazoline
(e.g. fluralaner)/kg of fish, up to about 5 mg isoxazoline (e.g.
fluralaner)/kg of fish,
for 5 to 15 days. The present disclosure also includes a use of an
isoxazoline,
composition, or medicated fish feed described herein for preventing or
treating
parasite infection in a plurality of fish, wherein the isoxazoline,
composition, or
medicated fish feed is for use at a daily dose comprising at least about 0.005
mg
isoxazoline (e.g. fluralaner)/kg of fish, up to about 5 mg isoxazoline (e.g.
fluralaner)/kg of fish, for 5 to 15 days. The present disclosure further
includes a
use of an isoxazoline, composition, or medicated fish feed described herein
for the
preparation of a medicament for preventing or treating parasite infection in a
plurality of fishõ wherein the medicament is for use at a daily dose
comprising at
least about 0.005 mg isoxazoline (e.g. fluralaner)/kg of fish, up to about 5
mg
isoxazoline (e.g. fluralaner)/kg of fish, for 5 to 15 days. The present
disclosure yet
further includes an isoxazoline, composition, or medicated fish feed described
herein for use for preventing or treating parasite infection in a plurality of
fish,
wherein the isoxazoline, composition, or medicated fish feed is for use at a
daily
dose comprising at least about 0.005 mg isoxazoline (e.g. fluralaner)/kg of
fish, up
to about 5 mg isoxazoline (e.g. fluralaner)/kg of fish, for 5 to 15 days. The
total
dose over a 10-day total treatment period would typically be 0.1 to 10 mg of
isoxazoline such as fluralaner per kg of fish. These same total dose amounts
may
be administered over a shorter dosage period such as 5, 6, 7, 8, or 9 days, or
a
longer dosage period, such as 11, 12, 13, 14, or 15 days. One of skill in the
art
would appreciate that a veterinary acceptable isoxazoline salt could
alternatively
be administered. The total isoxazoline such as fluralaner daily dose is
optionally
fed to the fish once a day. The total isoxazoline such as fluralaner daily
dose is
optionally fed to the fish as a series of divided daily doses optionally
comprising at
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least 2, 3, 4 or 5 feedings per day. After the total treatment period is
complete such
that the oral administration is complete for the desired number of days, the
fish are
typically held in the pen until isoxazoline residue decreases to a level that
the fish
are suitable for human consumption, e.g. less than 0.65ppm in the fish. The
method or use disclosed herein is effective to prevent or treat the parasite
(e.g. sea
lice) infection. Treatment typically achieves at least 90% removal for
significant
control of parasite (e.g. sea lice), however, optionally 100% removal of the
parasite
(e.g. sea lice) is achieved and it represents complete control of the
parasite.
[0024]
Isoxazolines are a class of inhibitors of gamma-aminobutyric acid
(GABA)- and glutamate-gated chloride channels in the nervous system of
invertebrates. Fluralaner is an isoxazoline approved by the U.S. Food and Drug
Administration (FDA) for treatment of flea and tick infestations in
terrestrial
animals. Fluralaner is sold under the tradename BravectoTM for the treatment
and
prevention of flea infestations, and the treatment and control of tick
infestations,
in cats and dogs. Fluralaner is also referred to as 4-[(5RS)-5-(3,5-
Di ch lorophenyI)-4, 5-d ihydro-5-(trifluoromethyl)-1 ,2-oxazol-3-y1]-N42-oxo-
2-
(2,2,2-trifluoroethylamino)ethyq-o-toluamide and Chemical Abstracts Service
(CAS) Registry Number 5864731-61-3.
[0025]
Isoxazoline afoxolaner is the active ingredient of the veterinary
medicinal products NexGardTM which is indicated for the treatment and
prevention of flea infestations, and the treatment and control of tick
infestations
in dogs and puppies. Afoxolaner is also referred to as 4-[(5RS)-5-(5-Chloro-
a,a,a-
trifluoro-m-tolyI)-4 , 5-d ihydro-5-(trifluoromethyl)-1 ,2-oxazol-3-y1]-N42-
oxo-2-
(2,2,2-trifluoroethylamino)ethyl]naphthalene-1-carboxamide and CAS Registry
Number 1093861-60-9.
[0026]
Isoxazoline lotilaner (tradename CredelioTM) is a veterinary drug
used to control fleas and ticks in dogs. Lotilaner is also referred to as 5-
[(55)-4,5-
Di hydro-5-(3,4, 5-trichl oropheny1)-5-(trifluoromethyl)-3-i soxazolyI]-3-
methyl-N 42-
oxo-2-[(2,2,2-trifl uoroethyl)am ino]ethy1]-2-thiophenecarboxamide and
CAS
Registry Number 1369852-71-0.
[0027]
In an embodiment, the isoxazoline comprises, consists essentially
of, or consists of fluralaner, afoxolaner, lotilaner, or a veterinary
acceptable salt
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of any of the foregoing. In another embodiment, the isoxazoline is fluralaner
or a
veterinary acceptable salt. In another embodiment, the isoxazoline is
afoxolaner
or a veterinary acceptable salt. In another embodiment, the isoxazoline is
lotilaner
or a veterinary acceptable salt.
[0028] The excipient can, for example, promote stability and
uniform mixing
with the active ingredient, or improve flavor and palatability and thus
increase
consumption of a medicated feed. Accordingly, in some embodiments, the
excipient comprises pork liver flavour, sucrose, maize starch, sodium lauryl
sulfate, disodium embonate monohydrate, magnesium stearate, aspartame,
glycerol, soya-bean oil, polyethylene glycol such as Macrogol 3350, vegetable
oil, or an approved pharmacopeia excipient. In an embodiment, the excipient
comprises one, two, three, or four approved pharmacopeia excipients. In an
embodiment, the approved pharmacopeia excipient comprises dextrose. In an
embodiment, the dextrose is a non-GMO dextrose. In an embodiment, the non-
GMO dextrose is Emdexe. Excipients such as carriers in composition or
medicated fish feed can help consumption of the composition or feed in fish.
In
an embodiment, the excipient comprises a carrier described herein.
[0029] A person skilled in the art can readily determine how
much
isoxazoline to mix an excipient such as a carrier to provide a pre-mix
composition
having a particular percentage of active isoxazoline. For example, in an
embodiment, the activity level of isoxazoline is first determined then the
isoxazoline is mixed with an excipient described herein, to give a pre-mix
comprising 40 wt% active isoxazoline which is 400 grams of active isoxazoline
per kilogram of pre-mix composition. In an embodiment, the concentration of
active isoxazoline in the composition is 10-90 wt%, 75-90 wt%, or 30-50 wt%,
optionally 35-45 wt%, optionally at least 10 wt%, based on the total weight of
the
composition. In an embodiment, the concentration of active isoxazoline in the
composition is 40 wt%. Medicated feed for fish can be prepared based on the
amount of feed that the fish are consuming each day or over the course of
treatment. For example, 10 mg of 40 wt% isoxazoline (e.g. fluralaner) per kg
of
fish dosage divided over 10 days of feeding, if used in a sea cage site, would
use
1 mg of active isoxazoline (e.g. fluralaner) per day for 10 days for 1 kg of
fish. A
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total biomass of 100,000 kg multiplied by 10 mg is 1,000,000 mg of active
isoxazoline (e.g. fluralaner) required over 10 days. This equals 1 kg of
active
isoxazoline (e.g. fluralaner) used. Since one kg of pre-mix composition may
provide 400 grams of isoxazoline (e.g. fluralaner), 2.5 kg of pre-mix is added
to
the fish feed.
[0030] The present disclosure provides feeding regimes that
result in
decreases sea lice infection and maintain good food uptake. In the methods and
uses of the present disclosure, an effective amount of the isoxazoline,
composition, or medicated fish feed is administered to the fish, optionally in
a
single administration, or preferably comprising a series of administrations.
For
example, the isoxazoline, composition, or medicated fish feed is administered
to
the fish over a dosage period of 5 to 15 days. Multiple feedings in fish (i.e.
a
divided daily dose) may be expected to maintain levels in the fish better than
a
single daily dose. Multiple dosages per day over a number of days can be used
to keep isoxazoline (e.g. fluralaner) levels high in the fish to kill the
parasite (e.g.
sea lice). Alternatively, for example, in farms having a colder water
temperature,
the dose is administered once per day. It will be appreciated by a person
skilled
in the art that a higher spike of medication can be obtained from such an
administration. In an embodiment, the isoxazoline, composition, or medicated
fish
feed is for oral use by feeding a divided dose in a plurality of feeds 1, 2,
3, 4, or
times daily, for 5 to 15 days. In an embodiment, the isoxazoline, composition,
or medicated fish feed is for oral use by feeding a divided dose in a
plurality of
feeds 1 time daily, for 5 to 15 days. In an embodiment, the isoxazoline,
composition, or medicated fish feed is for oral use by feeding a divided dose
in a
plurality of feeds 1 time daily, for 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15
days. In
an embodiment, the isoxazoline, composition, or medicated fish feed is for
oral
use by feeding a divided dose in a plurality of feeds 2 times daily, for 5 to
15 days.
In an embodiment, the isoxazoline, composition, or medicated fish feed is for
oral
use by feeding a divided dose in a plurality of feeds dose 2 times daily, for
5, 6,
7, 8, 9, 10, 11, 12, 13, 14, or 15 days. In an embodiment, the isoxazoline,
composition, or medicated fish feed is for oral use by feeding a divided dose
in a
plurality of feeds dose twice daily, for 10 days. In an embodiment, the
isoxazoline,
composition, or medicated fish feed is for oral use by feeding a divided dose
in a
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plurality of feeds 3 times daily, for 5 to 15 days. In an embodiment, the
isoxazoline,
composition, or medicated fish feed is for oral use by feeding a divided dose
in a
plurality of feeds 3 times daily, for 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15
days. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for oral
use by
feeding a divided dose in a plurality of feeds 4 times daily, for 5 to 15
days. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for oral
use by
feeding a divided dose in a plurality of feeds 4 times daily, for 5, 6, 7, 8,
9, 10, 11,
12, 13, 14, or 15 days. In an embodiment, the isoxazoline, composition, or
medicated
fish feed is for oral use by feeding a divided dose in a plurality of feeds 5
times daily,
for 5 to 15 days. In an embodiment, the isoxazoline, composition, or medicated
fish
feed is for oral use by feeding a divided dose in a plurality of feeds 5 times
daily, for
5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 days. In an embodiment, the daily
dose
comprises 0.01 mg isoxazoline (e.g. fluralaner)/kg of fish, up to about 2.5 mg
isoxazoline (e.g. fluralaner)/kg of fish. In an embodiment, the daily dose
comprises
about 0.07 mg isoxazoline (e.g. fluralaner)/kg of fish, up to about 0.7 mg
isoxazoline
(e.g. fluralaner)/kg of fish. In an embodiment, the daily dose comprises about
0.025
mg isoxazoline (e.g. fluralaner)/kg of fish, up to about 1 mg isoxazoline
(e.g.
fluralaner)/kg of fish. In an embodiment, the daily dose comprises about 0.025
mg
isoxazoline (e.g. fluralaner)/kg of fish, up to about 0.75 mg isoxazoline
(e.g.
fluralaner)/kg of fish. In an embodiment, the daily dose comprises about 0.05
mg
isoxazoline (e.g. fluralaner)/kg of fish, up to about 0.5 mg isoxazoline (e.g.
fluralaner)/kg of fish. In an embodiment, the daily dose comprises about 0.1
mg
isoxazoline (e.g. fluralaner)/kg of fish, up to about 0.25 mg isoxazoline
(e.g.
fluralaner)/kg of fish.
[0031] The skilled person can adjust dosage as necessary, for
example
adjusting on a per kilogram basis so the desired dose is measured and
administered on an amount per kilogram of fish. Optionally, 0.1 to 1 mg
isoxazoline
(e.g. fluralaner)/kg fish, such as 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.75,
0.8, 0.9, oil
mg isoxazoline (e.g. fluralaner)/kg of fish, is administered per day. In an
embodiment, the daily dose comprises about 0.005, 0.006, 0.007, 0.008, 0.009,
0.01, 0.02, 0.025, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.11, 0.12,
0.13,
0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.2, 0.21, 0.22, 0.23, 0.24, 0.25, 0.26,
0.27,
0.28, 0.29, 0.3, 0.31, 0.32, 0.33, 0.34, 0.35, 0.36, 0.37, 0.38, 0.39, 0.4,
0.41, 0.42,
11
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0.43, 0.44, 0.45, 0.46, 0.47, 0.48, 0.49, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75,
0.8, 0.85,
0.9, 0.95, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3,
2.4, 2.5, 2.6,
2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1,
4.2, 4.3, 4.4,
4.5, 4.6, 4.7, 4.8, 4.9, or 5.0 mg isoxazoline (e.g. fluralaner)/kg of fish.
In an
embodiment, the daily dose comprises about 0.01, 0.07, 0.1, 0.5, 0.7, 0.75, or
1
mg isoxazoline (e.g. fluralaner)/kg of fish. In an embodiment, the daily dose
comprises about 0.01 mg isoxazoline (e.g. fluralaner)/kg of fish. In an
embodiment, the daily dose comprises about 0.07 mg isoxazoline (e.g.
fluralaner)/kg of fish. In an embodiment, the daily dose comprises about 0.1
mg
isoxazoline (e.g. fluralaner)/kg of fish. In an embodiment, the daily dose
comprises
about 0.5 mg isoxazoline (e.g. fluralaner)/kg of fish. In an embodiment, the
daily
dose comprises about 0.7 mg isoxazoline (e.g. fluralaner)/kg of fish. In an
embodiment, the daily dose comprises about 0.75 mg isoxazoline (e.g.
fluralaner)/kg of fish. In an embodiment, the daily dose comprises about 1 mg
isoxazoline (e.g. fluralaner)/kg of fish. In an embodiment, the daily dose
comprises
about 0.01 mg isoxazoline (e.g. fluralaner)/kg of fish, for up to 10 days,
optionally
for 10 days. In an embodiment, the daily dose comprises about 0.07 mg
isoxazoline (e.g. fluralaner)/kg of fish, for up to 10 days, optionally for 10
days. In
an embodiment, the daily dose comprises about 0.1 mg isoxazoline (e.g.
fluralaner)/kg of fish, for up to 10 days, optionally for 10 days. In an
embodiment,
the daily dose comprises about 0.11 mg isoxazoline (e.g. fluralaner)/kg of
fish, for
up to 10 days, optionally for 10 days. In an embodiment, the daily dose
comprises
about 0.12 mg isoxazoline (e.g. fluralaner)/kg of fish, for up to 10 days,
optionally
for 10 days. In an embodiment, the daily dose comprises about 0.13 mg
isoxazoline (e.g. fluralaner)/kg of fish, for up to 10 days, optionally for 10
days. In
an embodiment, the daily dose comprises about 0.14 mg isoxazoline (e.g.
fluralaner)/kg of fish, for up to 10 days, optionally for 10 days. In an
embodiment,
the daily dose comprises about 0.15 mg isoxazoline (e.g. fluralaner)/kg of
fish, for
up to 10 days, optionally for 10 days. In an embodiment, the daily dose
comprises
about 0.16 mg isoxazoline (e.g. fluralaner)/kg of fish, for up to 10 days,
optionally
for 10 days. In an embodiment, the daily dose comprises about 0.17 mg
isoxazoline (e.g. fluralaner)/kg of fish, for up to 10 days, optionally for 10
days. In
an embodiment, the daily dose comprises about 0.17 mg isoxazoline (e.g.
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fluralaner)/kg of fish, for up to 10 days, optionally for 10 days. In an
embodiment,
the daily dose comprises about 0.18 mg isoxazoline (e.g. fluralaner)/kg of
fish, for
up to 10 days, optionally for 10 days. In an embodiment, the daily dose
comprises
about 0.19 mg isoxazoline (e.g. fluralaner)/kg of fish, for up to 10 days,
optionally
for 10 days. In an embodiment, the daily dose comprises about 0.2 mg
isoxazoline
(e.g. fluralaner)/kg of fish, for up to 10 days, optionally for 10 days. In an
embodiment, the daily dose comprises about 0.21 mg isoxazoline (e.g.
fluralaner)/kg of fish, for up to 10 days, optionally for 10 days. In an
embodiment,
the daily dose comprises about 0.22 mg isoxazoline (e.g. fluralaner)/kg of
fish, for
up to 10 days, optionally for 10 days. In an embodiment, the daily dose
comprises
about 0.23 mg isoxazoline (e.g. fluralaner)/kg of fish, for up to 10 days,
optionally
for 10 days. In an embodiment, the daily dose comprises about 0.24 mg
isoxazoline (e.g. fluralaner)/kg of fish, for up to 10 days, optionally for 10
days. In
an embodiment, the daily dose comprises about 0.25 mg isoxazoline (e.g.
fluralaner)/kg of fish, for up to 10 days, optionally for 10 days. In an
embodiment,
the daily dose comprises about 0.26 mg isoxazoline (e.g. fluralaner)/kg of
fish, for
up to 10 days, optionally for 10 days. In an embodiment, the daily dose
comprises
about 0.27 mg isoxazoline (e.g. fluralaner)/kg of fish, for up to 10 days,
optionally
for 10 days. In an embodiment, the daily dose comprises about 0.28 mg
isoxazoline (e.g. fluralaner)/kg of fish, for up to 10 days, optionally for 10
days. In
an embodiment, the daily dose comprises about 0.29 mg isoxazoline (e.g.
fluralaner)/kg of fish, for up to 10 days, optionally for 10 days. In an
embodiment,
the daily dose comprises about 0.3 mg isoxazoline (e.g. fluralaner)/kg of
fish, for
up to 10 days, optionally for 10 days. In an embodiment, the daily dose
comprises
about 0.31 mg isoxazoline (e.g. fluralaner)/kg of fish, for up to 10 days,
optionally
for 10 days. In an embodiment, the daily dose comprises about 0.32 mg
isoxazoline (e.g. fluralaner)/kg of fish, for up to 10 days, optionally for 10
days. In
an embodiment, the daily dose comprises about 0.33 mg isoxazoline (e.g.
fluralaner)/kg of fish, for up to 10 days, optionally for 10 days. In an
embodiment,
the daily dose comprises about 0.34 mg isoxazoline (e.g. fluralaner)/kg of
fish, for
up to 10 days, optionally for 10 days. In an embodiment, the daily dose
comprises
about 0.35 mg isoxazoline (e.g. fluralaner)/kg of fish, for up to 10 days,
optionally
for 10 days. In an embodiment, the daily dose comprises about 0.36 mg
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isoxazoline (e.g. fluralaner)/kg of fish, for up to 10 days, optionally for 10
days. In
an embodiment, the daily dose comprises about 0.37 mg isoxazoline (e.g.
fluralaner)/kg of fish, for up to 10 days, optionally for 10 days. In an
embodiment,
the daily dose comprises about 0.38 mg isoxazoline (e.g. fluralaner)/kg of
fish, for
up to 10 days, optionally for 10 days. In an embodiment, the daily dose
comprises
about 0.39 mg isoxazoline (e.g. fluralaner)/kg of fish, for up to 10 days,
optionally
for 10 days. In an embodiment, the daily dose comprises about 0.4 mg
isoxazoline
(e.g. fluralaner)/kg of fish, for up to 10 days, optionally for 10 days. In an
embodiment, the daily dose comprises about 0.41 mg isoxazoline (e.g.
fluralaner)/kg of fish, for up to 10 days, optionally for 10 days. In an
embodiment,
the daily dose comprises about 0.42 mg isoxazoline (e.g. fluralaner)/kg of
fish, for
up to 10 days, optionally for 10 days. In an embodiment, the daily dose
comprises
about 0.43 mg isoxazoline (e.g. fluralaner)/kg of fish, for up to 10 days,
optionally
for 10 days. In an embodiment, the daily dose comprises about 0.44 mg
isoxazoline (e.g. fluralaner)/kg of fish, for up to 10 days, optionally for 10
days. In
an embodiment, the daily dose comprises about 0.45 mg isoxazoline (e.g.
fluralaner)/kg of fish, for up to 10 days, optionally for 10 days. In an
embodiment,
the daily dose comprises about 0.46 mg isoxazoline (e.g. fluralaner)/kg of
fish, for
up to 10 days, optionally for 10 days. In an embodiment, the daily dose
comprises
about 0.47 mg isoxazoline (e.g. fluralaner)/kg of fish, for up to 10 days,
optionally
for 10 days. In an embodiment, the daily dose comprises about 0.48 mg
isoxazoline (e.g. fluralaner)/kg of fish, for up to 10 days, optionally for 10
days. In
an embodiment, the daily dose comprises about 0.49 mg isoxazoline (e.g.
fluralaner)/kg of fish, for up to 10 days, optionally for 10 days. In an
embodiment,
the daily dose comprises about 0.5 mg isoxazoline (e.g. fluralaner)/kg of
fish, for
up to 10 days, optionally for 10 days. In an embodiment, the daily dose
comprises
about 0.4 mg isoxazoline (e.g. fluralaner)/kg of fish, for up to 10 days,
optionally
for 10 days. In an embodiment, the daily dose comprises about 0.5 mg
isoxazoline
(e.g. fluralaner)/kg of fish, for up to 10 days, optionally for 10 days. In an
embodiment, the daily dose comprises about 0.7 mg isoxazoline (e.g.
fluralaner)/kg of fish, for up to 10 days, optionally for 10 days. In an
embodiment,
the daily dose comprises about 0.75 mg isoxazoline (e.g. fluralaner)/kg of
fish, for
up to 10 days, optionally for 10 days. In an embodiment, the daily dose
comprises
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about 1 mg isoxazoline (e.g. fluralaner)/kg of fish, for up to 10 days,
optionally for
days. In an embodiment, the isoxazoline, composition, or medicated fish feed
is for use at a total dose of about 0.025 mg isoxazoline (e.g. fluralaner)/kg
of fish,
up to about 75 mg isoxazoline (e.g. fluralaner)/kg of fish. In an embodiment,
the
isoxazoline, composition, or medicated fish feed is for use at a total dose of
about
0.05 mg isoxazoline (e.g. fluralaner)/kg of fish, up to about 25 mg
isoxazoline (e.g.
fluralaner)/kg of fish. In an embodiment, the isoxazoline, composition, or
medicated fish feed is for use at a total dose of about 0.1 mg isoxazoline
(e.g.
fluralaner)/kg of fish, up to about 10 mg isoxazoline (e.g. fluralaner)/kg of
fish. In
an embodiment, the isoxazoline, composition, or medicated fish feed is for use
at
a total dose of about 0.025, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1,
0.15, 0.2,
0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9,
0.95, 1,
1.05 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5,
2.6, 2.7, 2.8,
2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4,
4.5, 4.6, 4.7,
4.8, 4.9, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 11, 12, 13, 14, 15, 16,
17, 18, 19,
20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38,
39, 40,
41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 mg isoxazoline (e.g. fluralaner)/kg
of fish.
In an embodiment, the isoxazoline, composition, or medicated fish feed is for
use
for 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 days, for a total dose of about
0.5 mg
isoxazoline (e.g. fluralaner)/kg of fish. In an embodiment, the isoxazoline,
composition, or medicated fish feed is for use for 5, 6, 7, 8, 9, 10, 11, 12,
13,14, or
days, for a total dose of about 0.6 mg isoxazoline (e.g. fluralaner)/kg of
fish. In
an embodiment, the isoxazoline, composition, or medicated fish feed is for use
for
5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 days, for a total dose of about 0.7
mg
isoxazoline (e.g. fluralaner)/kg of fish. In an embodiment, the isoxazoline,
composition, or medicated fish feed is for use for 5, 6, 7, 8, 9, 10, 11, 12,
13,14, or
15 days, for a total dose of about 0.8 mg isoxazoline (e.g. fluralaner)/kg of
fish. In
an embodiment, the isoxazoline, composition, or medicated fish feed is for use
for
5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 days, for a total dose of about 0.9
mg
isoxazoline (e.g. fluralaner)/kg of fish. In an embodiment, the isoxazoline,
composition, or medicated fish feed is for use for 5, 6, 7, 8, 9, 10, 11, 12,
13,14, or
15 days, fora total dose of about 1 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use
for 5,
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6,7, 8,9, 10, 11, 12, 13, 14, or 15 days, for a total dose of about 1.1 mg
isoxazoline
(e.g. fluralaner)/kg of fish. In an embodiment, the isoxazoline, composition,
or
medicated fish feed is for use for 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15
days, fora
total dose of about 1.2 mg isoxazoline (e.g. fluralaner)/kg of fish. In an
embodiment,
the isoxazoline, composition, or medicated fish feed is for use for 5, 6, 7,
8, 9, 10,
11, 12, 13, 14, or 15 days, for a total dose of about 1.3 mg isoxazoline (e.g.
fluralaner)/kg of fish. In an embodiment, the isoxazoline, composition, or
medicated
fish feed is for use for 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 days, fora
total dose of
about 1.4 mg isoxazoline (e.g. fluralaner)/kg of fish. In an embodiment, the
isoxazoline, composition, or medicated fish feed is for use for 5, 6, 7, 8, 9,
10, 11,
12, 13, 14, or 15 days, for a total dose of about 1.5 mg isoxazoline (e.g.
fluralaner)/kg of fish. In an embodiment, the isoxazoline, composition, or
medicated
fish feed is for use for 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 days, fora
total dose of
about 1.6 mg isoxazoline (e.g. fluralaner)/kg of fish. In an embodiment, the
isoxazoline, composition, or medicated fish feed is for use for 5, 6, 7, 8, 9,
10, 11,
12, 13, 14, or 15 days, for a total dose of about 1.7 mg isoxazoline (e.g.
fluralaner)/kg of fish. In an embodiment, the isoxazoline, composition, or
medicated
fish feed is for use for 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 days, fora
total dose of
about 1.8 mg isoxazoline (e.g. fluralaner)/kg of fish. In an embodiment, the
isoxazoline, composition, or medicated fish feed is for use for 5, 6, 7, 8, 9,
10, 11,
12, 13, 14, or 15 days, for a total dose of about 1.9 mg isoxazoline (e.g.
fluralaner)/kg of fish. In an embodiment, the isoxazoline, composition, or
medicated
fish feed is for use for 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 days, fora
total dose of
about 2 mg isoxazoline (e.g. fluralaner)/kg of fish. In an embodiment, the
isoxazoline, composition, or medicated fish feed is for use for 5, 6, 7, 8, 9,
10, 11,
12, 13, 14, or 15 days, for a total dose of about 2.1 mg isoxazoline (e.g.
fluralaner)/kg of fish. In an embodiment, the isoxazoline, composition, or
medicated
fish feed is for use for 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 days, fora
total dose of
about 2.2 mg isoxazoline (e.g. fluralaner)/kg of fish. In an embodiment, the
isoxazoline, composition, or medicated fish feed is for use for 5, 6, 7, 8, 9,
10, 11,
12, 13, 14, or 15 days, for a total dose of about 2.3 mg isoxazoline (e.g.
fluralaner)/kg of fish. In an embodiment, the isoxazoline, composition, or
medicated
fish feed is for use for 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 days, for a
total dose of
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about 2.4 mg isoxazoline (e.g. fluralaner)/kg of fish. In an embodiment, the
isoxazoline, composition, or medicated fish feed is for use for 5, 6, 7, 8, 9,
10, 11,
12, 13, 14, or 15 days, for a total dose of about 2.5 mg isoxazoline (e.g.
fluralaner)/kg of fish. In an embodiment, the isoxazoline, composition, or
medicated
fish feed is for use for 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 days, fora
total dose of
about 2.6 mg isoxazoline (e.g. fluralaner)/kg of fish. In an embodiment, the
isoxazoline, composition, or medicated fish feed is for use for 5, 6, 7, 8, 9,
10, 11,
12, 13, 14, or 15 days, for a total dose of about 2.7 mg isoxazoline (e.g.
fluralaner)/kg of fish. In an embodiment, the isoxazoline, composition, or
medicated
fish feed is for use for 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 days, for a
total dose of
about 2.8 mg isoxazoline (e.g. fluralaner)/kg of fish. In an embodiment, the
isoxazoline, composition, or medicated fish feed is for use for 5, 6, 7, 8, 9,
10, 11,
12, 13, 14, or 15 days, for a total dose of about 2.9 mg isoxazoline (e.g.
fluralaner)/kg of fish. In an embodiment, the isoxazoline, composition, or
medicated
fish feed is for use for 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 days, fora
total dose of
about 3 mg isoxazoline (e.g. fluralaner)/kg of fish. In an embodiment, the
isoxazoline, composition, or medicated fish feed is for use for 5, 6, 7, 8, 9,
10, 11,
12, 13, 14, or 15 days, for a total dose of about 3.1 mg isoxazoline (e.g.
fluralaner)/kg of fish. In an embodiment, the isoxazoline, composition, or
medicated
fish feed is for use for 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 days, for a
total dose of
about 3.2 mg isoxazoline (e.g. fluralaner)/kg of fish. In an embodiment, the
isoxazoline, composition, or medicated fish feed is for use for 5, 6, 7, 8, 9,
10, 11,
12, 13, 14, or 15 days, for a total dose of about 3.3 mg isoxazoline (e.g.
fluralaner)/kg of fish. In an embodiment, the isoxazoline, composition, or
medicated
fish feed is for use for 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 days, fora
total dose of
about 3.4 mg isoxazoline (e.g. fluralaner)/kg of fish. In an embodiment, the
isoxazoline, composition, or medicated fish feed is for use for 5, 6, 7, 8, 9,
10, 11,
12, 13, 14, or 15 days, for a total dose of about 3.5 mg isoxazoline (e.g.
fluralaner)/kg of fish. In an embodiment, the isoxazoline, composition, or
medicated
fish feed is for use for 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 days, fora
total dose of
about 3.6 mg isoxazoline (e.g. fluralaner)/kg of fish. In an embodiment, the
isoxazoline, composition, or medicated fish feed is for use for 5, 6, 7, 8, 9,
10, 11,
12, 13, 14, or 15 days, for a total dose of about 3.7 mg isoxazoline (e.g.
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fluralaner)/kg of fish. In an embodiment, the isoxazoline, composition, or
medicated
fish feed is for use for 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 days, fora
total dose of
about 3.8 mg isoxazoline (e.g. fluralaner)/kg of fish. In an embodiment, the
isoxazoline, composition, or medicated fish feed is for use for 5, 6, 7, 8, 9,
10, 11,
12, 13, 14, or 15 days, for a total dose of about 3.9 mg isoxazoline (e.g.
fluralaner)/kg of fish. In an embodiment, the isoxazoline, composition, or
medicated
fish feed is for use for 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 days, fora
total dose of
about 4 mg isoxazoline (e.g. fluralaner)/kg of fish. In an embodiment, the
isoxazoline, composition, or medicated fish feed is for use for 5, 6, 7, 8, 9,
10, 11,
12, 13, 14, or 15 days, for a total dose of about 4.1 mg isoxazoline (e.g.
fluralaner)/kg of fish. In an embodiment, the isoxazoline, composition, or
medicated
fish feed is for use for 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 days, fora
total dose of
about 4.2 mg isoxazoline (e.g. fluralaner)/kg of fish. In an embodiment, the
isoxazoline, composition, or medicated fish feed is for use for 5, 6, 7, 8, 9,
10, 11,
12, 13, 14, or 15 days, for a total dose of about 4.3 mg isoxazoline (e.g.
fluralaner)/kg of fish. In an embodiment, the isoxazoline, composition, or
medicated
fish feed is for use for 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 days, fora
total dose of
about 4.4 mg isoxazoline (e.g. fluralaner)/kg of fish. In an embodiment, the
isoxazoline, composition, or medicated fish feed is for use for 5, 6, 7, 8, 9,
10, 11,
12, 13, 14, or 15 days, for a total dose of about 4.5 mg isoxazoline (e.g.
fluralaner)/kg of fish. In an embodiment, the isoxazoline, composition, or
medicated
fish feed is for use for 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 days, for a
total dose of
about 4.6 mg isoxazoline (e.g. fluralaner)/kg of fish. In an embodiment, the
isoxazoline, composition, or medicated fish feed is for use for 5, 6, 7, 8, 9,
10, 11,
12, 13, 14, or 15 days, for a total dose of about 4.7 mg isoxazoline (e.g.
fluralaner)/kg of fish. In an embodiment, the isoxazoline, composition, or
medicated
fish feed is for use for 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 days, fora
total dose of
about 4.8 mg isoxazoline (e.g. fluralaner)/kg of fish. In an embodiment, the
isoxazoline, composition, or medicated fish feed is for use for 5, 6, 7, 8, 9,
10, 11,
12, 13, 14, or 15 days, for a total dose of about 4.9 mg isoxazoline (e.g.
fluralaner)/kg of fish. In an embodiment, the isoxazoline, composition, or
medicated
fish feed is for use for 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 days, fora
total dose of
about 5 mg isoxazoline (e.g. fluralaner)/kg of fish. In an embodiment, the
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isoxazoline, composition, or medicated fish feed is for use at a daily dose
comprising at least about 0.005 mg isoxazoline (e.g. fluralaner)/kg of fish,
up to
about 5 mg isoxazoline (e.g. fluralaner)/kg of fish, for 5 to 15 days, for a
total dose
of about 0.025 mg isoxazoline (e.g. fluralaner)/kg of fish, up to about 75 mg
isoxazoline (e.g. fluralaner)/kg of fish. In an embodiment, the isoxazoline,
composition, or medicated fish feed is for use at a daily dose comprising at
least
about 0.01 mg isoxazoline (e.g. fluralaner)/kg of fish, up to about 5 mg
isoxazoline
(e.g. fluralaner)/kg of fish, for 10 days, for a total dose of about 0.1 mg to
about 50
mg isoxazoline (e.g. fluralaner)/kg of fish. In an embodiment, the
isoxazoline,
composition, or medicated fish feed is for use at a daily dose comprising
about
0.005 mg isoxazoline (e.g. fluralaner)/kg of fish, for 10 days, for a total
dose of
about 0.05 mg isoxazoline (e.g. fluralaner)/kg of fish. In an embodiment, the
isoxazoline, composition, or medicated fish feed is for use at a daily dose
comprising about 0.01 mg isoxazoline (e.g. fluralaner)/kg of fish, for 10
days, for
a total dose of about 0.1 mg isoxazoline (e.g. fluralaner)/kg of fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.02 mg isoxazoline (e.g. fluralaner)Ikg of fish,
for 10
days, for a total dose of about 0.2 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.03 mg isoxazoline (e.g. fluralaner)Ikg of fish,
for 10
days, for a total dose of about 0.3 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.04 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 0.4 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.05 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 0.5 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.06 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 0.6 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.07 mg isoxazoline (e.g. fluralaner)Ikg of fish,
for 10
days, for a total dose of about 0.7 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
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embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.08 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 0.8 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.09 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 0.9 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.1 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 1 mg isoxazoline (e.g. fluralaner)fkg of fish.
In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.11 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 1.1 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.12 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 1.2 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.13 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 1.3 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.14 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 1.4 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.15 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 1.5 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.16 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 1.6 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.17 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 1.7 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.18 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 1.8 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
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embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.19 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 1.9 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.2 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 2 mg isoxazoline (e.g. fluralaner)/kg of fish.
In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.21 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 2.1 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.22 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 2.2 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.23 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 2.3 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.24 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 2.4 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.25 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 2.5 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.26 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 2.6 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.27 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 2.7 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.28 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 2.8 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.29 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 2.9 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
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embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.3 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 3 mg isoxazoline (e.g. fluralaner)/kg of fish.
In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.31 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 3.1 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.32 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 3.2 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.33 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 3.3 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.34 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 34 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.35 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 3.5 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.36 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 3.6 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.37 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 3.7 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.38 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 3.8 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.39 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 3.9 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.4 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 4 mg isoxazoline (e.g. fluralaner)/kg of fish.
In an
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embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.41 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 4.1 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.42 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 4.2 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.43 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 4.3 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.44 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 4.4 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.45 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 4.5 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.46 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 4.6 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.47 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 4.7 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.48 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 4.8 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.49 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 4.9 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.5 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 5 mg isoxazoline (e.g. fluralaner)/kg of fish.
In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.2 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 5
days, for a total dose of about 1 mg isoxazoline (e.g. fluralaner)/kg of fish.
In an
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embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.167 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 6
days, for a total dose of about 1 mg isoxazoline (e.g. fluralaner)/kg of fish.
In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.143 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 7
days, for a total dose of about 1 mg isoxazoline (e.g. fluralaner)/kg of fish.
In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.125 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 8
days, for a total dose of about 1 mg isoxazoline (e.g. fluralaner)fkg of fish.
In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.111 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 9
days, for a total dose of about 1 mg isoxazoline (e.g. fluralaner)/kg of fish.
In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.1 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 1 mg isoxazoline (e.g. fluralaner)/kg of fish.
In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.091 mg isoxazoline (e.g. fluralaner)/kg of fish,
for
11 days, for a total dose of about 1 mg isoxazoline (e.g. fluralaner)/kg of
fish. In
an embodiment, the isoxazoline, composition, or medicated fish feed is for use
at
a daily dose comprising about 0.083 mg isoxazoline (e.g. fluralaner)/kg of
fish, for
12 days, for a total dose of about 1 mg isoxazoline (e.g. fluralaner)/kg of
fish. In
an embodiment, the isoxazoline, composition, or medicated fish feed is for use
at
a daily dose comprising about 0.077 mg isoxazoline (e.g. fluralaner)/kg of
fish, for
13 days, for a total dose of about 1 mg isoxazoline (e.g. fluralaner)/kg of
fish. In
an embodiment, the isoxazoline, composition, or medicated fish feed is for use
at
a daily dose comprising about 0.071 mg isoxazoline (e.g. fluralaner)/kg of
fish, for
14 days, for a total dose of about 1 mg isoxazoline (e.g. fluralaner)/kg of
fish. In
an embodiment, the isoxazoline, composition, or medicated fish feed is for use
at
a daily dose comprising about 0.067 mg isoxazoline (e.g. fluralaner)/kg of
fish, for
15 days, for a total dose of about 1 mg isoxazoline (e.g. fluralaner)/kg of
fish. In
an embodiment, the isoxazoline, composition, or medicated fish feed is for use
at
a daily dose comprising about 0.11 mg isoxazoline (e.g. fluralaner)/kg of
fish, for
days, for a total dose of about 1.1 mg isoxazoline (e.g. fluralaner)/kg of
fish. In
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an embodiment, the isoxazoline, composition, or medicated fish feed is for use
at
a daily dose comprising about 0.12 mg isoxazoline (e.g. fluralaner)/kg of
fish, for
days, for a total dose of about 1.2 mg isoxazoline (e.g. fluralaner)/kg of
fish. In
an embodiment, the isoxazoline, composition, or medicated fish feed is for use
at
a daily dose comprising about 0.13 mg isoxazoline (e.g. fluralaner)/kg of
fish, for
10 days, for a total dose of about 1.3 mg isoxazoline (e.g. fluralaner)/kg of
fish. In
an embodiment, the isoxazoline, composition, or medicated fish feed is for use
at
a daily dose comprising about 0.14 mg isoxazoline (e.g. fluralaner)/kg of
fish, for
10 days, for a total dose of about 1.4 mg isoxazoline (e.g. fluralaner)/kg of
fish. In
an embodiment, the isoxazoline, composition, or medicated fish feed is for use
at a
daily dose comprising about 0.15 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 1.5 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.2 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 2 mg isoxazoline (e.g. fluralaner)/kg of fish.
In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.25 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 2.5 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.3 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 3 mg isoxazoline (e.g. fluralaner)/kg of fish.
In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.35 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 3.5 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.4 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 4 mg isoxazoline (e.g. fluralaner)/kg of fish.
In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.45 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 4.5 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.5 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 5 mg isoxazoline (e.g. fluralaner)/kg of fish.
In an
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embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.55 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 5.5 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.6 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 6 mg isoxazoline (e.g. fluralaner)/kg of fish.
In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.65 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 6.5 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.7 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 7 mg isoxazoline (e.g. fluralaner)/kg of fish.
In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 0.75 mg isoxazoline (e.g. fluralaner)/kg of fish,
for 10
days, for a total dose of about 7.5 mg isoxazoline (e.g. fluralaner)/kg of
fish. In an
embodiment, the isoxazoline, composition, or medicated fish feed is for use at
a
daily dose comprising about 1 mg isoxazoline (e.g. fluralaner)/kg of fish, for
10
days, fora total dose of about 10 mg isoxazoline (e.g. fluralaner)/kg of fish.
[0032] Optionally, the isoxazoline, composition, or
medicated fish feed is for
oral use by feeding a divided dose in a plurality of feeds once every 1 to 4
days for
up to 15 days.
[0033] The isoxazoline, composition, or medicated fish feed
disclosed
herein can be used to treat or prevent parasite, e.g. sea lice infection in a
fish,
including any economically important fish such as salmonid. In an embodiment,
the fish is carp, eel, trout, whitefish, salmon, bream, roach, rudd, chub,
sole, plaice,
halibut, Japanese yellowtail (Seriola quinqueradiata), Japanese eel (Anguilla
japonica), red sea bream (Pa gurus major), sea bass (Dicen trarchus labrax),
grey
mullet (Mug//us cephalus), pompano, giltbread sea bream (Sparus auratus),
Tilapia species, East Atlantic peacock wrasse (Symphodus tinca), cichlid
species,
such as, for example, plagioscion, channel catfish. In an embodiment, the fish
is
salmonid. In an embodiment, the salmonid is a salmon or trout. In an
embodiment,
the salmonid is Salmo salar, Oncorhynchus tshawytscha, Oncorhynchus kisutch,
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Oncorhynchus keta, Oncorhynchus gorbuscha, Oncorhynchus nerka, or
Oncorhynchus mykiss. In an embodiment, each salmonid has an average mass
of less than or equal to 5 kg, optionally an average mass of from 25 g to 5
kg. The
isoxazoline, isoxazoline-containing composition, or isoxazoline-containing
medicated fish feed can treat parasites, e.g. sea lice infections. In an
embodiment,
the parasite is Copepodae (cyclops; fish-lice) with the genera Ergasilus,
Bomolochus, Chondracanthus, Caligus (e.g. Caligus curtus), Lepeophtheirus
(e.g.
L. salmonis), Elythrophora, Dichelesthium, Lamproglena, Hatschekia,
Leposphilus, Pseudocycnus, Lemaea, Lemaeocera, PenneIla, Achtheres,
Basanistes, Salmincola, Brachiella, Epibrachiella, Pseudotracheliastes,
Argulus,
and Ceratothoa (e.g. Ceratothoa gaudichaudii). In an embodiment, the parasite
is
a sea louse.
[0034] The life cycle of sea lice begins at the nauplius
stage when these
planktonic nauplii cannot swim directionally against the water current but
drift
passively. The free swimming nauplii then moult into the infective copepodid
stage. At this stage, the copepodid has the ability to clasp onto the host
tissue,
and then it undergoes a moult to the immobile chalimus stages in the life
cycle. At
the chalimus stages, the sea lice are attached to the fish before becoming pre-
adults or mobile stage. At the pre-adult stage, the sea lice are able to move
around
on the surface of the fish and can also swim in the water. Two pre-adult
stages
are followed by the fully mature adult phase, at which point the sea lice have
reached sexual maturity and the gravid females produce egg-strings from which
the planktonic nauplii hatch, repeating the life cycle. In an embodiment, the
sea
louse is at any stage of life-cycle. In an embodiment, the sea louse is pre-
adult
sea lice, copepodites, chalimus, adult sea lice, or gravid female lice. In an
embodiment, the sea louse is pre-adult sea louse. In an embodiment, the sea
louse is copepodites. In an embodiment, the sea louse is chalimus. In an
embodiment, the sea louse is adult sea lice. In an embodiment, the sea louse
is
gravid female lice. In an embodiment, the sea louse is Lepeophtheirus or
Caligus,
optionally the Lepeophtheirus is Lepeophtheirus salmonis, optionally the
Caligus
is Caligus clemensi, Caligus elongatus, or Caligus rogercresseyi.
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[0035] In an embodiment, after use, the fish are held in a
pen prior to
harvesting for the minimum time necessary for the isoxazoline residue
(optionally
fluralaner residue) to be reduced to a level that the fish is suitable for
human
consumption, optionally until residue testing on fish determines that the
isoxazoline residue in the fish is below 0.65 parts per million.
[0036] In an embodiment, the fish are harvested, optionally
wherein the fish
are harvested 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 21, 22, 23, 24, 0r25
days
after the use is completed, optionally when isoxazoline residue (optionally
fluralaner residue) in the fish is below 0.65 parts per million.
[0037] The present disclosure also includes a medicated fish
feed
comprising fish feed granules or pellets coated or mixed with a composition
comprising an isoxazoline and at least one excipient.
[0038] The present disclosure also includes a pre-mix
composition for
medicated fish feed comprising an isoxazoline and at least one excipient.
[0039] The selection of a suitable means to mix the
isoxazoline and the
excipient can be made by a person skilled in the art. Then, for example, to
formulate and administer the composition, a suitable amount (e.g. 5 kilograms)
of
the isoxazoline (e.g. fluralaner) containing pre-mix composition is mixed with
a
suitable amount (e.g. 5,000 kilogram) of fish feed and fed to the fish as the
sole
diet for, for example, 10 days. Optionally, a dry pre-mix composition is mixed
in
fish oil then applied to the fish feed, optionally in a vacuumed fat
applicator. A
vacuumed fat applicator is a commonly used apparatus in large fish feed making
plants.
[0040] The fish feed granules or pellets can be any suitable
fish feed granules
or pellets, the selection of which can be made by a person skilled in the art.
[0041] In an embodiment, the method or use further
comprises, after
administration or use, holding the fish in a pen prior to harvesting for the
minimum
time necessary for the isoxazoline residue (optionally fluralaner residue) to
be
reduced to a level that the fish is suitable for human consumption, optionally
until
residue testing on fish determines that the isoxazoline residue in the fish is
below
0.65 parts per million. In an embodiment, the method or use further comprises
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harvesting the fish, optionally 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 21,
22, 23, 24,
or 25 days after the administration or use is completed, optionally when
isoxazoline residue (optionally fluralaner residue) in the fish is below 0.65
parts
per million. It will be appreciated by a person skilled in the art that
residue levels
acceptable for harvest and therefore holding times may vary between different
countries. It will also be appreciated that lower water temperatures typically
increase excretion times of medications in fish (i.e. at lower water
temperatures
fish typically excrete lower amounts of medication per day) so longer holding
periods would be expected at lower temperatures. The selection of suitable
holding times and residue levels can be made by a skilled person.
III. Medicated Fish Feed
[0042] The present inventor discloses that feeding
isoxazoline (e.g.
fluralaner) to fish in the form of a suitable medicated fish feed decreases
parasite
infestations (e.g. sea lice infections) and maintains good food uptake at
doses of
0.01 to 0.1 mg isoxazoline (e.g. fluralaner)/kg fish per day. The medicated
fish
feed used in the studies of the present disclosure was prepared by mixing feed
pellet with powder formulation of fluralaner, and sealed with fish oil to help
coat it
onto the pellets of the salmon feed.
[0043] Accordingly, the present disclosure also includes
medicated fish
feed comprising fish feed granules or pellets coated or mixed with a
composition
comprising an isoxazoline and a carrier. It will be appreciated by a person
skilled
in the art that embodiments relating, for example, to the isoxazoline and the
carrier
in the medicated fish feeds of the present disclosure can be varied as
detailed
herein for the methods and uses of the present disclosure. In an embodiment,
the
medicated fish feed further comprises krill meal.
[0044] The following non-limiting Examples are illustrative
of the present
disclosure:
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EXAMPLES
Example 1: Feed Trial
I. Method
In-feed treatment of sea lice infested Atlantic salmon (Salmo salar) with an
isoxazoline class of compound
[0045] Atlantic Salmon (Salmo salar) with an average weight
of 116 grams
were allocated to 80-liter aquariums, ten fish per aquarium. Fish were
infected
with pre-adult sea lice (Lepeophtheirus salmonis) for five days, at which time
treatment with fluralaner began. The isoxazoline class of compounds include
fluralaner, afoxolaner and lotilaner. The test compound in this example was
fluralaner, which was tested at 3 doses. Experimental diets were prepared by
top
dressing 3 mm floating feed pellets with the powder formulation of fluralaner
and
sealed with fish oil, this medicated feed was fed to the fish twice daily for
ten days.
Uneaten medicated feed pellets were removed and counted each day to estimate
average dosage received by the fish. Fish were examined for sea lice 5 days
following the end of the treatment period.
II. Results
Intake results of the feed
Table 1. Dose achieved in treatment groups based on uneaten feed
Compound % of target dose achieved
Nominal dose mg
fluralaner/kg of fish/per 1.0 0.1
0.01
day (for over 10 days)
Control 100 100 100
Fluralaner 22 100 98
[0046] As shown in Table 1, the higher dosage had more
reduction in feed
intake. Optionally, one could potentially add feed attractants such as krill
meal to
the diet to minimize reduction in feed intake.
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Efficacy results on the sea lice
Table 2. Reduction of Sea Lice after Fluralaner Treatment
Summary of Reduction of total sea lice after
Fluralaner Treatment
Nominal dose
mg fluralaner/kg
of fish/per day 1.0 0.1 0.01
(for over 10
days)
Total dose mg
fluralaner / kg of
1 0.1
fish (over 10
days)
Reduction of
total sea lice (/0) 0 0 0
Control
Reduction of
total sea lice (%) 100 100 79
Fl uralaner
[0047] Five days after the 10-day treatment period the
average sea lice
count on the control group was 5.6 pre-adult lice. The overall reduction of
sea lice
with the different dosages is shown in Table 2. Inventor showed that two of
the
dosages, at 1.0 mg and 0.1 mg fluralaner/kg fish/per day for over 10 days,
100%
of sea lice were removed from the fish. These were surprisingly good results.
[0048] One of skill in the art will obtain similar results
to those obtained for
fluralaner with use of afoxolaner and lotilaner, based on the disclosure in
this
application. Accordingly, the medicated feed of the present disclosure is
useful to
deliver other isoxazolines for example, for sea louse control in salmonid and
other
fish. Other types of isoxazolines are useful in the formulations and doses
described herein to prevent and treat parasite infection in fish.
Example 2: Dosages Regime Trials
[0049] Experiments using large salmon cages up to 100 meters
in
circumference containing close to 5 million liters of water that is constantly
being
replaced by ocean tidal currents do present field situation where sea lice
will need
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to be treated. To get close to this field situation, eight large tanks each
containing
1,200 liters of water with a flow through water supply were used at a research
facility for dosage regime trials.
Preparation and Calculation of the concentrations of test material
[0050] Medicated fish feed containing fluralaner was
prepared for dose
rates of 0.2, 0.1, and 0.05 mg fluralaner / kg of fish biomass per day with a
5%
overage to account for loss. Materials were prepared to be sufficient to feed
70
fish per group, with the fish having an average base weight of 433.7 g, thus
the
biomass of each group was 30.359 kg. Therefore, the expected daily 0.6% food
consumption was estimated as 0.182 kg/day. For example, the target
concentration of the active ingredient in the medicated feed pellets for the
dosage
to give 0.1 mg of fluralaner, per kilogram of fish per day, plus a 5% overage
for
anticipated loss in mixing, was 17.5 mg of active in 1,000 g of medicated feed
pellets. Fish oil was included at 0.75% or 7.5 g oil / kg pellets.
Table 3. Preparation of Medicated Fish Feed
Preparation of Mediated Fish Feed
Dose 0
06 0.2 0.1 0.
(mg/kg/day)
(Control)
Ave fish wgt (g) 433.7 433.7 433.7
433.7
Number 70 70 70
70
Biomass (kg) 30.359 30.359 30.359
30.359
Dose
0.2 0.1 0.05 0
(mg/kg/day)
Overage rate 0.05 0.05 0.05
0.05
Feed rate (0.6%) 0.006 0.006 0.006
0.006
Daily feed (kg) 0.182 0.182 0.182
0.182
Target
Concentration 35 17.5 8.75 0
(mg/kg feed)
Premix
Concentration 134.68 134.68 134.68
134.68
(mg/g)
Total Premix
0.260 0.130 0.065 0
(g/kg feed)
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Days of Feed
(available for 55 55 55 55
feeding)
Batch Feed (kg) 10.018 10.018 10.018
10.018
Batch premix (g) 2.604 1.302 0.651
0.000
Fish oil inclusion
7.5 7.5 7.5 7.5
rate g/kg Feed
Batch oil g 75.14 75.14 75.14
75.14
Results
[0051] Atlantic Salmon (Salmo salar) with an average weight
of 434 grams
were allocated to large salmon tanks (1,200-liter tanks), at thirty-five fish
per tank
(two tanks for each dosage and control). Fish were infected with sea lice
(Lepeophtheirus salmonis) at the copepodite stage five days before treatment
with
fluralaner began. Fluralaner was tested at 3 doses (0.2, 0.1, 0.05 mg
fluralaner /
kg fish / per day and control with no fluralaner). Medicated fish feed
prepared
above was fed to the fish twice daily for ten days. At 11 and 20 days
following the
end of the treatment period, fish were examined for sea lice counts. At 11
days
after the end of the 10-day treatment period, the average mobile sea lice
(i.e. pre-
adults) count on the control group was 41 mobile preadult lice. At 20 days
after
treatment ended, the control group averaged 32.9 mobile preadult lice. As
shown
in Table 4 and Table 5, daily dose of at least 0.1 fluralaner / kg fish for
over 10
days (or at least 1 mg fluralaner / kg fish per entire treatment) was
effective in
reducing sea lice infection on salmon.
Table 4. Summary of Sea Lice Reductions at 11 Days Post Treatment (Mobile
Preadult Stage)
Large Trial
Dose mg
0
fluralaner/ kg of 0.2 0.1 0.05
(Control)
fish/per day
Total mg
fluralaner / kg of
2 1 0.5 0
fish over 10
days
Reduction of
95% 89% 49% 0%
sea lice (%) at
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day 11 post
treatment
Table 5. Summary of Sea Lice Reductions at 20 Days Post Treatment (Mobile
Preadult Stage)
Large Trial
Dose mg
0
fluralaner / kg of 0.2 0.1 0.05
(Control)
fish/per day
Total mg
fluralaner / kg of
2 1 0.5 0
fish over 10
days
Reduction of
mobile sea lice
95% 86% 48% 0%
(%) at day 20
post treatment
[0052] These results, and the results from Example 1, show
that fluralaner
treatment is effective in reducing sea lice infection on salmon, including sea
lice at
different life-cycle stages.
[0053] While the present disclosure has been described with
reference to what
are presently considered to be the preferred examples, it is to be understood
that the
present disclosure is not limited to the disclosed examples To the contrary,
the
present disclosure is intended to cover various modifications and equivalent
arrangements included within the spirit and scope of the appended claims.
[0054] All publications, patents and patent applications are
herein
incorporated by reference in their entirety to the same extent as if each
individual
publication, patent or patent application was specifically and individually
indicated
to be incorporated by reference in its entirety. Where a term in the present
disclosure is found to be defined differently in a document incorporated
herein by
reference, the definition provided herein is to serve as the definition for
the term.
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