A SUPPLEMENT

FIELD

The present invention relates to a dietary supplement suitable for aquatic animals and methods associated therewith.

BACKGROUND

Aquatic animals, particularly those farmed for meat production, are prone to stress related conditions, often a result of confinement, the intensity of the population, and/or deficiencies in diet. These conditions can result in increased disease mortality, loss of or reduced disease resistance, darkening of the skin, reduced meat quality including preferential accumulation of fat on the gut rather than in the muscle, and reduced fresh shelf life, for example.

Products including pharmaceuticals, foods, and dietary supplements have been suggested to be of benefit in supporting health and dampening some of the negative effects associated with stress and farming of land animals. These include natural feed supplements comprising extracts of berries including blackcurrants. However, such products are not formulated for use in an aquatic environment. If used in an aquatic environment, a number of problems may arise which may affect the effective delivery of the product to the aquatic animals. For example, the product may be unstable in water or the solubility of a product in water may be undesirable; the product may be too soluble or not soluble enough making it unavailable to the aquatic animals or giving rise to difficulties in ensuring enough product or a desired dosage of a product is supplied. Typically aquaculture feed products must be insoluble. This can be problematic for many natural products, for example, as they are soluble in water. In addition, many products may be unpalatable to the aquatic animals.

Feed supplements have been used in aquaculture. These include supplements comprising astaxanthin or beta-glucan. Such products are said to promote weight gain, and in the case of astaxanthin, impart desirable colour to fish flesh, but they are not known to provide any other benefits, and the fish may still suffer from one or more of the problems referred to above. In addition, the cost of including such supplements in a farming setting is typically very expensive. Antibiotics are also often used in aquaculture. As noted above, aquatic animals are prone to disease, particularly in intensive farming environments. This means that many farms are reliant on the use of antibiotics to reduce disease resistance. However, there is consumer concern that the use of antibiotics in animals intended for human consumption poses health risks for humans. Accordingly, there is continued pressure to reduce the reliance on antibiotics and offer alternative natural solutions. Reliance on antibiotics can also increase the cost of farming fish.

OBJECT

It is an object of the present invention to provide a dietary supplement and associated methods which overcome or ameliorate at least one of the disadvantages of the prior art, or to at least provide the public with a useful alternative.

STATEMENT OF INVENTION

In one broad aspect, the present invention provides a composition comprising one or more anthocyanins, processed whole blackcurrants and/or a blackcurrant extract, wherein the composition is suitable for use in an aquatic environment.

The composition may be referred to as a supplement herein.

In one embodiment, the supplement comprises processed whole blackcurrants and/or a blackcurrant extract.

In one embodiment, the supplement comprises one or more of whole blackcurrants, a blackcurrant puree, a blackcurrant pomace, blackcurrant juice or juice concentrate, and a blackcurrant extract.

In one embodiment, a supplement of the invention comprises from approximately 0.05% to approximately 3.0% anthocyanins (w/w). In one embodiment, a supplement of the invention comprises from approximately 0.05% to approximately 0.4% anthocyanins

(w/w). In one embodiment, a supplement of the invention comprises from approximately 0.08% to approximately 3.0% anthocyanins (w/w). In one embodiment, the supplement comprises a blackcurrant concentrate comprising from approximately 800mg to approximately 3000mg anthocyanin/lOOg concentrate. In one particular embodiment, the supplement comprises a blackcurrant concentrate comprising 1220mg anthocyanin/lOOg of concentrate.

In another particular embodiment, the supplement comprises a blackcurrant pomace comprising from approximately 50mg to approximately 400mg anthyocyanin/lOOg of pomace. In one embodiment the supplement comprises a blackcurrant pomace comprising approximately 50mg to approximately 300mg anthocyanins/lOOg of pomace. In one particular embodiment, the supplement comprises a blackcurrant pomace comprising approximately 80mg anthocyanins/lOOg of pomace.

In another particular embodiment, the supplement comprises a dried blackcurrant pomace comprising from approximately lOOmg to approximately 2700mg anthyocyanin/lOOg of dried pomace.

In one embodiment, a single dose of the supplement comprises from approximately 0.5mg to approximately lOmg anthocyanin(s) per kg of body weight. In one particular embodiment, a single dose of the supplement comprises from approximately 0.5 to approximately 3.5mg per kg of body weight. In one preferred embodiment a single dose of the supplement comprises approximately 2.4mg per kg of body weight. In another preferred embodiment a single dose of the supplement comprises approximately 1.2mg per kg of body weight. In another preferred embodiment a single dose of the supplement comprises approximately 0.8mg per kg of body weight.

In another broad aspect, the invention provides a feed for an aquatic animal comprising processed whole blackcurrants, an extract of blackcurrants and/or a supplement as herein before described. In one embodiment, the feed is in pellet form. In one embodiment, the pellet is a glazed pellet. In another embodiment, the feed is in an encapsulated form. In another

embodiment, the feed is in gel form. In another embodiment, the feed is in flake form. In one embodiment, the feed comprises from approximately 0.1 % to approximately 30% blackcurrants/extract/supplement by weight of total feed. In one embodiment, the feed comprises from approximately 0.5% to approximately 30%

blackcurrants/extract/supplement by weight of total feed. In a preferred embodiment, the feed comprises from approximately 0.1 % to approximately 5.0%

blackcurrants/extract/supplement by weight of total feed. In one particular embodiment, the feed comprises from approximately 2.0% to approximately 20%

blackcurrants/extract/supplement. In one embodiment, the feed is prepared to comprise an anthocyanin concentration of from approximately 0.01% to approximately 0.25% by weight of the total feed. In one embodiment, the feed is prepared to comprise an anthocyanin concentration of from approximately 0.01% to approximately 0.1% by weight of the total feed. In one particular embodiment, the feed is prepared to comprise an anthocyanin concentration of approximately 0.015%.

In another broad aspect, the invention provides the use of blackcurrants and/or an extract of blackcurrants in the manufacture of a medicament, dietary supplement or feed for delivery to an aquatic animal.

In another broad aspect, the invention provides the use of blackcurrants and/or an extract of blackcurrants as a fish feed ingredient.

In another broad aspect, the invention provides the use of blackcurrants and/or an extract of blackcurrants in the manufacture of a medicament, dietary supplement or feed for: substantially maintaining or improving the health of an aquatic animal; substantially maintaining or improving skin colour of an aquatic animal; substantially maintaining or improving flesh colour of an aquatic animal; substantially maintaining or improving meat taste and texture of an aquatic animal; substantially maintaining or improving the smell of an aquatic animal; substantially maintaining or extending fresh shelf life of a harvested aquatic animal; substantially maintaining or extending cooked shelf life of a harvested aquatic animal; substantially maintaining or improving fat content in the flesh of an aquatic animal; substantially maintaining or improving oil quality of an aquatic animal; substantially maintaining or improving the firmness of the flesh of an aquatic animal; substantially reducing gaping in fillets of an aquatic animal; substantially improving weight of useable flesh in an aquatic animal; substantially reducing the reliance of an aquatic animal on antibiotics; substantially reducing the reliance of an aquatic animal on astaxanthin supplementation; and/or, substantially preventing or reducing accumulation of fat around the gut of an aquatic animal.

In another broad aspect, the invention provides the use of one or more anthocyanins in the manufacture of a medicament, dietary supplement or feed for delivery to an aquatic animal. Preferably, the one or more anthocyanins are from blackcurrant. Preferably, the anthocyanins are a combination of anthocyanins from blackcurrant.

Use of one or more anthocyanins as a fish feed ingredient. Preferably, the one or more anthocyanins are from blackcurrant. Preferably, the anthocyanins are a combination of anthocyanins from blackcurrant.

In another broad aspect, the invention provides the use of one or more anthocyanins in the manufacture of a medicament or dietary supplement for: substantially maintaining or improving the health of an aquatic animal; substantially maintaining or improving skin colour of an aquatic animal; substantially maintaining or improving flesh colour of an aquatic animal; substantially maintaining or improving meat taste and texture of an aquatic animal; substantially maintaining or improving the smell of an aquatic animal;

substantially maintaining or extending fresh shelf life of a harvested aquatic animal;

substantially maintaining or extending cooked shelf life of a harvested aquatic animal; substantially maintaining or improving fat content in the flesh of an aquatic animal;

substantially maintaining or improving oil quality of an aquatic animal; substantially maintaining or improving the firmness of the flesh of an aquatic animal; reducing gaping in fillets of an aquatic animal; substantially improving weight of useable flesh in an aquatic animal; substantially reducing the reliance of an aquatic animal on antibiotics; substantially reducing the reliance of an aquatic animal on astaxanthin supplementation; and/or, substantially preventing or reducing accumulation of fat around the gut of an aquatic animal. Preferably, the one or more anthocyanins are from blackcurrant. Preferably, the anthocyanins are a combination of anthocyanins from blackcurrant. In another broad aspect, the invention provides a method for: substantially maintaining or improving the health of an aquatic animal; substantially maintaining or improving skin colour of an aquatic animal; substantially maintaining or improving flesh colour of an aquatic animal; substantially maintaining or improving meat taste and texture of an aquatic animal; substantially maintaining or improving the smell of an aquatic animal;

substantially maintaining or extending fresh shelf life of a harvested aquatic animal;

substantially maintaining or extending cooked shelf life of a harvested aquatic animal; substantially maintaining or improving fat content in the flesh of an aquatic animal;

substantially maintaining or improving oil quality of an aquatic animal; substantially maintaining or improving the firmness of the flesh of an aquatic animal; substantially reducing gaping in fillets of an aquatic animal; substantially improving weight of useable flesh in an aquatic animal; substantially reducing the reliance of an aquatic animal on antibiotics; substantially reducing the reliance of an aquatic animal on astaxanthin supplementation; and/or, substantially preventing or reducing accumulation of fat around the gut of an aquatic animal; wherein the method comprises delivering a supplement and/or feed of one of the previous aspects of the invention to an aquatic animal.

In another broad aspect, the invention provides a method for: substantially maintaining or improving the health of an aquatic animal; substantially maintaining or improving skin colour of an aquatic animal; substantially maintaining or improving flesh colour of an aquatic animal; substantially maintaining or improving meat taste and texture of an aquatic animal; substantially maintaining or improving the smell of an aquatic animal;

substantially maintaining or extending fresh shelf life of a harvested aquatic animal;

substantially maintaining or extending cooked shelf life of a harvested aquatic animal; substantially maintaining or improving fat content in the flesh of an aquatic animal;

substantially maintaining or improving oil quality of an aquatic animal; substantially maintaining or improving the firmness of the flesh of an aquatic animal; substantially reducing gaping in fillets of an aquatic animal; substantially improving weight of useable flesh in an aquatic animal; substantially reducing the reliance of an aquatic animal on antibiotics; substantially reducing the reliance of an aquatic animal on astaxanthin supplementation; and/or, substantially preventing or reducing accumulation of fat around the gut of an aquatic animal; wherein the method comprises delivering one or more anthocyanins to an aquatic animal. Preferably, the one or more anthocyanins are from blackcurrant. Preferably, the anthocyanins are a combination of anthocyanins from blackcurrant. In a particular embodiment, supplement and/or feed is delivered to the animal in sufficient quantity to provide a dose of approximately 0.5 to approximately lOmg anthocyanin per kg of body weight. In one particular embodiment, supplement and/or feed is delivered to the animal in sufficient quantity to provide a dose of approximately 0.5 to approximately 3.5mg per kg of body weight. In one preferred embodiment, the supplement and/or feed is delivered to the animal in sufficient quantity to provide a dose of approximately 2.4mg per kg of body weight. In another preferred embodiment, the supplement and/or feed is delivered to the animal in sufficient quantity to provide approximately 1.2mg per kg of body weight. In another preferred embodiment, the supplement and/or feed is delivered to the animal in sufficient quantity to provide a dose of approximately 0.8mg per kg of body weight.

In one embodiment, the blackcurrants, blackcurrant extract, supplement, feed and/or one or more anthocyanins are delivered to the animal daily. In another embodiment, they are delivered to the animal every second day. In another embodiment, they are delivered twice a week.

In one embodiment the aquatic animal is a fish, mollusc, crustacean, or an echinoderm. In one embodiment, the aquatic animals are farm raised. The invention may also be said broadly to consist in the parts, elements and features referred to or indicated in the specification of the application, individually or collectively, in any or all combinations of two or more of said parts, elements or features, and where specific integers are mentioned herein which have known equivalents in the art to which the invention relates, such known equivalents are deemed to be incorporated herein as if individually set forth. FIGURES

These and other aspects of the present invention, which should be considered in all its novel aspects, will become apparent from the following description, which is given by way of example only, with reference to the accompanying figures, in which:

Figure 1: Shows skin darkening and discolouration and slender body shape of control as compared to supplemented fish.

Figure 2: Shows heavy fat accumulation on the gut of the control fish compared to very little in supplemented fish.

Figure 3: Shows heavy fat accumulation on the gut of the control fish compared to very little in supplemented fish (closer view). Figure 4: Shows skin darkening and discolouration and slender body shape of control as compared to supplemented fish (closer view).

PREFERRED EMBODIMENT(S)

The following is a description of the present invention, including preferred embodiments thereof, given in general terms. The invention is further elucidated from the disclosure given under the heading "Examples" herein below, which provides experimental data supporting the invention.

The inventors have conducted trials and surprisingly identified that a fish supplemented with a blackcurrant composition has a number of health and production benefits in farmed fish including improved colour, improved meat taste and texture, extended fresh shelf life of harvested fish and less accumulation of fat around the gut, for example. They have also surprisingly observed that supplementation reduces the need to administer antibiotics and/or astaxanthin to fish. They have also observed a number of other unexpected benefits in supplementing fish feed in accordance with the invention, as is described elsewhere herein. Accordingly, they believe there is value in use of blackcurrant products as dietary supplements to improve the health and production quality of fish and other aquatic animals, particularly those farmed in an intensive environment.

The trials were conducted over a number of months. Some preferred compositions of the invention were developed as a result of these trials. In formulating the compositions the inventors had to consider a number of significant problems and often competing requirements including, for example, one or more of formulation for effective delivery in an aquatic environment, dose size, delivery of the desired amount of active ingredients, palatability, shelf life, bioavailability and side-effects. The high solubility of blackcurrant juice and desirable components in blackcurrants was of particular concern, as there was no guarantee that the inventors would be able to achieve a stable product and effective delivery of an appropriate dose of the product to the fish.

While the inventors contemplate the use of processed whole blackcurrants or an extract of whole blackcurrants could be used as a supplement on its own, they have found that there is significant benefit in formulating the blackcurrant component, for example with other ingredients, to overcome the above issues. Use of unformulated blackcurrent component may lead to cloudy water, poor availability of the blackcurrant including anthocyanins to the fish, poor delivery of the desired amount of the blackcurrant including anthocyanins.

While the trials conducted by the inventors focussed on specific species of fish and crustaceans, the inventors believe the invention is applicable to any number of aquatic animals. As used herein, the term "aquatic animal(s)" should be taken to include any animal living in an aquatic environment including but not limited to fish, molluscs, crustaceans, echinoderms. In one embodiment, the aquatic animals are farm raised.

By way of example only, the invention may find particular application in farming of snapper, salmon, carp, tilapia, European seabass, catfish, cod, cobia, trout, barramundi, sturgeon, tuna, shrimp, prawns, clams, oysters, mussels, scallops and Ayu.

The invention provides a dietary supplement for an aquatic animal. In one embodiment, the supplement comprises processed whole blackcurrants. In one embodiment, the supplement comprises a blackcurrant extract. It should be appreciated that processed whole blackcurrants and blackcurrant extract(s) may be used in combination.

"Processed whole blackcurrants" are blackcurrants which have been treated in some way with minimal and preferably no removal or loss of any part of the fruit. The blackcurrants are preferably processed in a way that retains a substantial proportion of the activity or benefits of the anthocyanin(s) present therein. Whole blackcurrants may be dried and/or pureed, for example. A "blackcurrant extract" is a product derived from blackcurrants which has been separated from one or more of the natural components of the whole fruit; for example, one or more individual compounds, more complex mixtures of compounds, a particular tissue, seeds, skin and/or fluid of the fruit. The extract may comprise one or more compounds or ingredients and preferably includes one or more anthocyanins. The extract may also optionally contain one or more polyphenols and/or vitamin C. In one embodiment, the extract is obtained from the skin, pulp and/or seeds of the fruit; for example, it is a pomace. In another embodiment, the extract comprises the juice of the fruit.

Unless the context requires otherwise, a "blackcurrant component" or "blackcurrant ingredient" as may be referred to herein is intended to cover processed whole

blackcurrants, a blackcurrant extract and/or mixtures thereof. It may also include a supplement of the invention.

In particular embodiments, the supplement comprises one or more of whole blackcurrants (for example, fresh or dried), a blackcurrant puree (for example, as a liquid or dried), a blackcurrant pomace (for example, with or without seeds in any form, which may also be dried), blackcurrant juice or juice concentrate (for example, as a liquid or dried), a blackcurrant extract (for example, as a liquid or dried). It will be appreciated that "dried" ingredients may be dried using any appropriate means, including, for example, air drying, vacuum drying and freeze drying. In particular embodiments, vacuum drying or freeze drying is used. Where the blackcurrant ingredient is dried it may also be processed into a powder, as is the case in some preferred embodiments of the invention. In a preferred embodiment, the blackcurrants are chosen from the Ben Ard and Ben Rua varieties, preferably grown in New Zealand. However, other varieties could be used.

The blackcurrants of use in the invention preferably have an anthocyanin concentration of from approximately 200mg/100g of fruit to approximately 400-600mg/100g.

The size of a single dose of a supplement of the invention will vary depending on factors such as the aquatic animal to which it is being supplied, the growth rate of the aquatic animal and the environment in which it is contained. However, generally the supplement will be supplied to deliver approximately 0.5 to approximately lOmg anthocyanin per kg of body weight with each dose or feeding. In one particular embodiment, a single dose of the supplement comprises from approximately 0.5 to approximately 3.5mg per kg of body weight. In other embodiments, a single dose of the supplement comprises from approximately lmg to approximately 9mg anthocyanin(s) per kg of body weight, from approximately 2mg to approximately 8mg anthocyanin(s) per kg of body weight, or from approximately 3mg to approximately 7mg anthocyanin(s) per kg of body weight. Further, by way of example only, in the case of a fish of approximately lOg in size (a small fish) the composition may be supplied to deliver approximately 0.05 mg anthocyanin per dose, and in the case of a fish of approximately 4kg (a large fish) the composition may be supplied to deliver approximately lOmg anthocyanin per dose. In one embodiment, the supplement is designed to provide a dose of approximately 1 to 1.5mg, and preferably approximately 1.2mg, anthocyanin per kg body weight per feeding. This is of particular use with farmed snapper. In another embodiment, the supplement is designed to provide a dose of approximately 2 to 3mg anthocyanins per kg of body weight and in a particular embodiment, approximately 2.4mg per kg of body weight. In another embodiment, the supplement is designed to provide a dose of approximately 0.05 mg to 1.2mg anthocyanins per kg of body weight and in a particular embodiment approximately 0.8mg/kg of body weight. These embodiments are of particular use with farmed prawns. In a preferred embodiment, a supplement of the invention comprises from approximately 0.05% to approximately 3.0% anthocyanins (w/w). In one particular embodiment, a supplement of the invention comprises a pomace comprising from approximately 0.05% to approximately 0.4% anthocyanins (w/w). In another particular embodiment, a supplement of the invention comprises a pomace comprising from approximately 0.08% to

approximately 3.0% anthocyanins (w/w). In other embodiments, a supplement of the invention comprises from approximately 0.1% to approximately 2.5%, from approximately 0.2% to approximately 2.0%, from approximately 0.3% to approximately 1.5%, or from approximately 0.4% to approximately 1.0%.

In one particular embodiment, the supplement comprises a blackcurrant concentrate comprising from approximately 800mg to approximately 3000mg anthocyanin/lOOg of concentrate. In another particular embodiment, the supplement comprises a blackcurrant pomace comprising from approximately 50mg to approximately 400mg anthyocyanin/lOOg of pomace. In one embodiment, the supplement comprises a blackcurrant pomace comprising from approximately 50mg to approximately 300mg anthyocyanin/lOOg of pomace. In one particular embodiment, the supplement comprises a blackcurrant pomace comprising approximately 80mg anthocyanins/ lOOg of pomace.

In another particular embodiment, the supplement comprises a dried blackcurrant pomace comprising from approximately lOOmg to approximately 2700mg anthyocyanin/lOOg of dried pomace.

It should be appreciated that a dietary supplement of the invention may include ingredients in addition to those mentioned herein before. For example, they may include fillers, carriers, binding agents, extenders, flavour enhancers, colourants, compounds which increase shelf life, gelling agents, encapsulating agents, other fruits including fruit extracts, and/or compounds which may provide additional health benefits, among other ingredients.

Processed whole black currants and blackcurrant extracts may be made using any number of standard techniques known in the art, including methods for pureeing, forming a pomace, extraction of one or more compounds, drying (including, for example, air drying, vacuum drying, freeze drying) and grinding. However, by way of example only, a blackcurrant concentrate suitable for use in the invention may be prepared from whole blackcurrants, fresh or frozen, using the following methodology: 1) tipping blackcurrants; 2) milling; 3) mash heating; 4) pressing/filtration to separate skins/seeds from juice; 5) separation centrifugation of juice; 6) pastuerisation; 7) optionally, storing; 8) filtering (MF 1.0 micron); 9) evaporation to form a concentrate.

By way of further example, a blackcurrant pomace of use in the invention may be prepared using the following methodology: 1) tipping blackcurrants; 2) milling; 3) mash heating; 4) pressing/filtration to separate juice from skins/seeds (pomace).

In a preferred embodiment of the invention, the whole blackcurrants and blackcurrant extract are used in a dried form; for example, a blackcurrant concentrate, a blackcurrant pomace, blackcurrant puree, whole blackcurrants or the like are dried. In a particularly preferred embodiment, vacuum drying is used. The inventors have surprisingly found that by using vacuum drying they can reduce the amount of the blackcurrant ingredient in the supplement of the invention while still achieving the same positive effects. For example, in one embodiment, the inventors have been able to reduce the amount of blackcurrant ingredient from 0.6% to 0.5% of the supplement. Any appropriate vacuum drying procedure may be used. However, by way of example, a proprietary process of the YahiroYahiro company may be used. Typically, the drying time for whole blackcurrants, blackcurrant concentrate, blackcurrant extract is approximately 18 to 36 hours and for a blackcurrant pomace approximately 12-24 hours, for one drying load of approximately 650kg of inwards (starting) weight.

In another embodiment, the dried blackcurrant ingredients of the supplement are ground. In a preferred embodiment, the dried ingredients are ground to between approximately 100 and approximately 450 micron. In preferred embodiments, the ingredients are ground to between approximately 100 to 425 microns. In one particular embodiment, the ingredients are ground to approximately 250 microns. When the dried blackcurrant ingredients are milled to within these ranges the inventors observe benefits in terms of improved dispersion throughout a supplement or feed formulation of the invention. Where a supplement of the invention contains more than one ingredient, the various ingredients may be combined or mixed in the desired ratio, using any known methodology. Alternatively, by way of further example, the blackcurrant ingredient could be added to a pre-made fish supplement using techniques such as soaking and glazing as described later herein.

The supplements of the invention may be in any appropriate form for use in an aquatic environment. Based on the information provided herein, skilled persons would readily appreciate appropriate supplement forms. However, by way of example, the supplements of the invention may be processed so they are in flake form, pellet form, insoluble particulate form, gel form or an encapsulated form (including soft gels). In one

embodiment, where the supplements are provided in the form of pellets, they are glazed. The inventors contemplate these forms are particularly suited to delivery of the supplement to an aquatic environment, in a manner which is palatable and stable for a sufficient period of time to allow an aquatic animal to ingest at least a sufficient amount of the desired dose, preferably substantially the whole dose, in its normal feeding period.

The supplement of the invention is preferably packed into containers which provide a gas barrier. In a preferred embodiment the container is a gas barrier foil pouch. The supplement may be packed into the container with or without inert gas flushing to prevent oxidative or photo degradation of the anthocyanin(s). Such packing may help ensure an extended shelf life of the supplement. In one embodiment, the supplement is formulated for delivery to an aquatic animal in combination with feed for the aquatic animal(s). Accordingly, the invention also provides feed for an aquatic animal, the feed comprising a dietary supplement as herein described.

A variety of feeds and feed ingredients are suitable for aquatic animals, as will be appreciated by persons of ordinary skill in the art. However, by way of example only, the feed may comprise one or more of Wheat Germ Meal, Wheat Red Dog, Wheat Bran, Wheat Middlings, Wheat Mill Run, Corn Gluten Feed, Corn Gluten Meal, Rice Groats, Rice Bran with Germ, Rice Polishings, Oat Groats, Soybean Meals, Cottonsead Meals, Peanut Meal, Hydrolyzed Poultry Feathers, Meat and Bone Meal, Fish Meals, vegetable protein, vitamins, minerals and Blood Meal.

In certain embodiments, ready-made feeds may be used and combined with a supplement of the invention. Such feeds include for example, those manufactured and supplied by Biomar (Denmark), EWOS AS (Norway) and Skretting (Norway).

A feed of the invention may also comprise any number of additional ingredients including but not limited to carriers, fillers, diluents, extenders, flavour enhancers, colourants, compounds which increase shelf life, encapsulation agents, gelling agents, other fruits including fruit extracts, and/or compounds which may provide additional health benefits. The feed may also comprise standard additives, such as oils, which allow the feed to be adapted to sink or float, depending on the farming or other environment it is intended for. In one embodiment, the feed ingredients and processed whole blackcurrants, a blackcurrant extract, and/or a supplement of the invention are combined in a ratio of from

approximately 0.1% to approximately 30% blackcurrants/extract/supplement to feed ingredients. In another embodiment, the feed ingredients and processed whole

blackcurrants, a blackcurrant extract, and/or a supplement of the invention are combined in a ratio of from approximately 0.5% to approximately 30%

blackcurrants/extract/supplement to feed ingredients. In one particular embodiment, the feed ingredients and processed whole blackcurrants, a blackcurrant extract, and/or a supplement of the invention are combined in a ratio of from approximately 2% to approximately 20% blackcurrants/extract/supplement to feed ingredients. In another particular embodiment, the feed ingredients and processed whole blackcurrants, a blackcurrant extract, and/or a supplement of the invention are combined in a ratio of from approximately 0.1% to approximately 5% blackcurrants/extract/supplement to feed ingredients. In other embodiments, the feed ingredients and processed whole blackcurrants, a blackcurrant extract, and/or a supplement of the invention are combined in a ratio of from approximately 0.1 % to approximately 20%, from approximately 0.5% to

approximately 20%, from approximately 1.0% to approximately 10%, from approximately 5.0% to approximately 20%, or from approximately 5.0% to 10%

blackcurrants/extract/supplement to feed ingredients. In one particular embodiment, the feed is prepared to comprise an anthocyanin

concentration of from approximately 0.01% to approximately 0.25% by weight of the total feed. In one embodiment, the feed is prepared to comprise an anthocyanin concentration of from approximately 0.01% to approximately 0.1% by weight of the total feed. In one particular embodiment, the feed is prepared to comprise an anthocyanin concentration of approximately 0.015%. In other embodiments, the feed is prepared to comprise an anthocyanin concentration of from approximately 0.01% to approximately 0.2%, from approximately 0.01% to approximately 0.1%, from approximately 0.01% to approximately 0.05%, or from approximately 0.01 % to approximately 0.02%.

The feeds of the invention may be in any appropriate form for use in an aquatic

environment. In certain embodiments, the feed of the invention may be processed so it is in flake form, pellet form, insoluble particulate form, gel form or an encapsulated form (including soft gels). In one embodiment, where the feed is provided in the form of pellets, they are glazed. These forms are particularly suited to delivery of the feed to an aquatic environment. The inventors have found that supplements in pellet form, flake form, glazed pellet form are particularly suited to delivery of the supplement to an aquatic environment, in a manner which is palatable and stable for a sufficient period of time to allow an aquatic animal to ingest at least a sufficient amount of the desired dose, preferably substantially the whole dose, in its normal feeding period.

The feed may be prepared using a number of different techniques allowing for the combination of two or more ingredients. According to well known principles, the feed may also be manufactured in a number of different sizes, adapted to the size of the fish and/or farming or other environment in which it is to be used.

Persons of ordinary skill in the art will readily appreciate methodology for preparing supplements and feeds of the invention, for example: pellets, flakes, gels, capsules and insoluble particulate feed. However, by way of general example, pellets can be made using the equipment and methodology of Andritz Inc. By way of further example, encapsulated products can be made using a rotary die encapsulation process. This encapsulation process has been described as a form/fill/seal process. Briefly, two flat ribbons of shell material are manufactured and brought together on a twin set of rotating dies. The dies contain recesses in the desired size and shape, which cut out the ribbons into a two dimensional shape, and form a seal around the outside. At the same time a pump delivers a precise dose of fill material through a nozzle incorporated into a filling wedge whose tip sits between the two ribbons in between two die pockets at the point of cut out. The wedge is heated to facilitate the sealing process. The wedge injection causes the two flat ribbons to expand into the die pockets, giving rise to the three dimensional finished product. After encapsulation, the product may be dried for two days to two weeks depending on the product. In the case of a softgel, for example, the shell may comprise gelatin (or one or more other polymers based on, for example, starch and carrageenan), water, opacifier and a plasticiser such as glycerin and/or sorbitol(s).

In a further example, micro-encapsulation techniques may be employed to prepare supplements and feeds of the invention including both physical, physico-chemical and chemical techniques. Such techniques include, for example, pan coating, centrifugal extrusion, vibrational nozzle, spray drying, interfacial polycondensation, interfacial cross- linking, in-situ polymerization, matrix polymerization, ionotropic gelation and

coacervation. In one embodiment, a feed is made or otherwise supplied and a blackcurrant extract in liquid form is allowed to absorb into the feed. In one embodiment, the liquid extract is a juice concentrate and it is diluted approximately 1: 1 with water and then allowed to absorb into the pre-made feed. In other embodiments, each of the individual feed ingredients and the supplement of the invention are mixed and processed together to form the feed.

In another embodiment, the feed is in pellet form and is produced by grinding and mixing standard fish food ingredients together with a blackcurrant ingredient in accordance with the invention. In one preferred embodiment, the blackcurrant ingredient is a dried blackcurrant pomace, preferably made by freeze or vacuum drying. Water is added and then the paste is extruded through holes in a metal plate and pellets are cut to the desired size. Pellets are then dried and oils added where desired. In a further embodiment, the feed is a glazed pellet. The feed is produced by mixing standard fish feed ingredients together and forming the pellets as described above. The pellets are then coated or glazed with a blackcurrant compound according to standard procedures. The blackcurrant compound used for the glaze may comprise any of the blackcurrant ingredients or supplements described herein before as being of use in the invention. However, in preferred embodiments of the invention, the glaze comprises one or more of: a dried blackcurrant pomace powder, a dried blackcurrant puree powder, a dried whole blackcurrant powder, a dried blackcurrant concentrate powder, or a blackcurrant concentrate; alone or in combination with water, solutions or wetting agents.

In another embodiment, the feed is in the form of dried flakes. The flakes may be purchased or produced from standard ingredients using known methodology. The flakes can then been soaked in a blackcurrant solution. The soaked flakes are then dried. The flakes may be processed to be of any desirable size. The blackcurrant solution used may comprise any of the blackcurrant ingredients or supplements described herein before as being of use in the invention. However, in preferred embodiments of the invention, the solution comprises one or more of: a dried blackcurrant pomace powder, a dried blackcurrant puree powder, a dried whole blackcurrant powder, a dried blackcurrant concentrate powder or a blackcurrant concentrate. The solution may comprise water, or other appropriate solutions or wetting agents.

Alternatively, the flakes may be manufactured by mixing desired ingredients (such as standard fish feed ingredients) and a blackcurrant ingredient in accordance with the invention and then processed according to standard techniques.

The feed of the invention is preferably packed into containers which provide a gas barrier. In a preferred embodiment the container is a gas barrier foil pouch. The feed may be packed into the container with or without inert gas flushing to prevent oxidative or photo degradation of the anthocyanin(s). Preferably, the supplements and feeds of the invention will be packaged in bulk moisture and light barrier protective packaging. This may help ensure an extended shelf life. In use, the supplements and feed may be administered to the aquatic animal in any appropriate way. In one preferred embodiment, the supplement is provided in the form of a feed. The supplements and feeds of the invention may be used as alternatives to existing feeds for aquatic animals, for the purposes of substantially maintaining or improving the health of an aquatic animal; substantially maintaining or improving skin colour of an aquatic animal; substantially maintaining or improving flesh colour of an aquatic animal;

substantially maintaining or improving meat taste and texture of an aquatic animal;

substantially maintaining or improving the smell of an aquatic animal; substantially maintaining or extending fresh shelf life of a harvested aquatic animal; substantially maintaining or extending cooked shelf life of a harvested aquatic animal; substantially maintaining or improving fat content in the flesh of an aquatic animal; substantially maintaining or improving oil quality of an aquatic animal; substantially maintaining or improving the firmness of the flesh of an aquatic animal; substantially reducing gaping in fillets of an aquatic animal; substantially improving weight of useable flesh in an aquatic animal; substantially reducing the reliance of an aquatic animal on antibiotics; substantially reducing the reliance of an aquatic animal on astaxanthin supplementation; and/or, substantially preventing or reducing accumulation of fat around the gut of an aquatic animal.

The term "maintaining" refers to substantially preserving, or substantially preventing deterioration of, a particular condition, state or quality existing in an animal or population of animals.

The "health" of an aquatic animal encompasses reduced disease mortality, increased disease resistance and/or increased immunity to one or more pathogens, infections or diseases. Skin colour and flesh colour can vary in an aquatic animal for a number of reasons, including general health and environmental conditions. Darkening skin colour may be undesirable. Reference to "improving skin colour" and "improving flesh colour" and like terms are intended to include brightening or lightening of the colour. Meat texture can vary in aquatic animals for a number of reasons. The market may demand fish having substantially consistent texture, substantially no gaping of flesh, and substantially firmer flesh, or otherwise fish meat which more closely resembles the texture of fresh aquatic animals harvested from natural environments. Reference to "improving meat quality and/or texture" is intended to include improving one or more of these features which can be assessed visually.

The taste of meat is an important aspect in the market for aquatic animals. Taste of the meat can be strongly influenced by the environmental conditions in which the animal is grown, among other things. "Improving meat taste" is therefore intended to include improving the taste according to market needs and/or to more closely resemble the taste of aquatic animals farmed in clean harvesting environments or natural environments. Farmed aquatic animals often have a strong and unappealing smell. Accordingly,

"improving the smell" of an aquatic animal is intended to mean that this characteristic farm fish odour is reduced, made less noticable, substantially removed, or the smell of the aquatic animals more closely resembles the smell of aquatic animals farmed in clean harvesting environments or natural environments.

"Improving fat content in the flesh of an aquatic animal" is intended to mean that the quality of the fat content in the flesh of the aquatic animal is improved relative to the standard quality of the fat content in farmed aquatic animals. For example, the fat flavour is noticeably improved when the flesh is eaten and the appearance of the fat in the animal is noticeably improved.

"Extending shelf life" should be construed broadly to include any lengthening of the time from harvest until the animal is no longer suitable for consumption or commercial sale. In one embodiment, the invention extends the fresh shelf life of a harvested aquatic animal by up to 48 hours; for example, at least 5 hours, 10 hours, 20 hours, 30 hours, 40 hours or 48 hours. In another embodient, the invention extends the cooked shelf life of a harvested aquatic animal by up to 72 hours; for example, at least 10 hours, 20 hours, 30 hours, 40 hours, 50 hours, 60 hours, or 72 hours. "Improving oil quality of an aquatic animal" is intended to mean that the natural oils of the flesh of the aquatic animal are of a higher quality or higher nutritional level. "Improving weight of useable flesh" is intended to mean that the ratio of meat that is suitable for consumer or other use compared to total body weight has increased, at least to some degree. In one embodiment, the weight of the usuable flesh is increased by between approximately 10% to approximately 40%, for example approximately 30%. "Reducing the reliance on antibiotics" should be taken broadly to include any reduction in the level of antibiotics used compared to when the supplement or feed of the invention is not used. In certain embodiments, the reduction in antibiotic use is at least approximately 5%, approximately 10%, approximately 20%, approximately 30%, approximately 40%, approximately 50%. In some embodiments, the use of antibiotics is substantially eliminated.

"Reducing the reliance on astaxanthin" should be taken broadly to include any reduction in the level of astaxanthin used compared to when the supplement or feed of the invention is not used. In certain embodiments, the reduction in astaxanthin use is at least

approximately 5%, approximately 10%, approximately 20%, approximately 30%, approximately 40%, approximately 50%. In some embodiments, the use of astaxanthin is substantially eliminated.

As mentioned previously herein, fat can accumulate around the gut of an aquatic animal. This may be undesirable. "Preventing or reducing accumulation of fat around the gut" of an aquatic animal should be taken to mean that the supplement or feed of the invention will substantially prevent additional fat accumulating around the gut of the animal upon supplementation or that the fat around the gut will be at least reduced to some level compared to an animal not fed the supplement or feed of the invention. In some embodiments, the fat around the gut is substantially eliminated. In another embodiment, the supplements and feeds of the invention may be used in the manufacture of medicaments, compositions and/or feeds for a use as herein before described. The supplements and/or feeds of the invention may be provided to the aquatic animal at any appropriate time and frequency. Feeding rates may vary depending on factors such as the growth rate of the aquatic animal, the type of aquatic animal, the environmental conditions (such as season and temperature), and the stage of development of the animal. Skilled persons will be readily able to adjust feeding rates to different conditions, without undue experimentation. Typically the supplement/feed of the invention will be provided at the regular feeding time for the animal. The supplement may be provided in a single dose, or a number of divided doses, to achieve a desired daily dose. In certain embodiments, the supplements/feeds are provided to the animals daily, every second day, or twice a week. It may be desirable to provide the supplements/feeds to the animal more frequently during the active growth period (for example when water temperatures are warmer) and less frequently at other times. By way of example, in winter the animal may be fed twice a week and as the weather warms the feeding may be increased from 3 to 4 times per week to once per day in the peak of summer. It is preferable that the animal is on a regular maintenance programme of the supplements/feeds.

The dosage provided to individual animals may vary having regard to the species, size of the animal, feeding programme, intensity of farming and the like. However, by way of example, the animal is fed the supplement/feed to achieve a dose per feeding of anthocyanin(s) from approximately 0.5mg kg of body weight to approximately lO.Omg/kg of body weight. In one particular embodiment, the animal is fed the supplement/feed to achieve a dose per feeding of approximately 0.5 to approximately 3.5mg anthocyanin per kg of body weight. In another embodiment, the animal is fed the supplement/feed to achieve a dose of from approximately l.Omg/kg of body weight to approximately

5.0mg/kg of body weight.

In other embodiments, a dose comprises from approximately lmg to approximately 9mg anthocyanin(s) per kg of body weight, from approximately 2mg to approximately 8mg anthocyanin(s) per kg of body weight, or from approximately 3mg to approximately 7mg anthocyanin(s) per kg of body weight.

In one embodiment, a dose comprises approximately 1 to 1.5mg, and preferably approximately 1.2mg, anthocyanin per kg body weight per feeding. This is of particular use with farmed snapper. In another embodiment, a dose comprises approximately 2 to 3mg anthocyanins per kg of body weight and in a particular embodiment, approximately 2.4mg per kg of body weight. In another embodiment, a dose comprises approximately 0.05 mg to 1.2mg anthocyanins per kg of body weight and in a particular embodiment approximately 0.8mg/kg of body weight. These embodiments are of particular use with farmed prawns.

EXAMPLES Example 1:

A study was completed to test the efficacy of the supplements to support and improve the health of fish being farmed by aquaculture.

Trial: The initial trial was designed for pen raised red snapper with a calculated dose requirement of 0.6mg anthocyanin per feeding.

Blackcurrant Concentrate 1220mg anthocyanin /100g concentrate

Feed weight per fish per feeding 4.17g The blackcurrant concentrate was prepared from Ben Ard blackcurrants using the methodology outlined hereinbefore.

To incorporate the required dose into pre-made fish pellets, 1.5kg of concentrate was diluted 1 :1 with water and allowed to absorb into 125kg of the feed pellets, enough sufficient for one feeding of the trial pen of 30,000 fish.

One pen of 30,000 fish received the supplemented feed. A second pen of 30,000 identical fish received no blackcurrant supplementation. The fish were fed twice per week at the start of the trial through the winter while the water temperature remained <13°C. Digestion requires 3 - 3.5 days in such cold conditions. When water temperatures increased above 13°C the fish were fed daily. This is the active growth period. The trial was run for approximately 12 months.

Sample fish were taken from the control pen and the trial pen, killed, bled and placed on ice. They were then shipped to a chef for analysis. Shipping and transport to the chef required 22hrs before the fish were presented.

Results: Surprisingly, fish fed the of the invention showed: improvement in the colour of the skin (a common defect in pen raised fish is darkening of the skin), retention of fat content in the flesh, extended fresh shelf life of harvested fish, improvement in the colour, texture and taste of the flesh, increased the yield of usable flesh from the fish and reductions in gut weight. This is evidenced by the Figures and the comments from a top sushi chef in Japan.

Sushi Chef Comments:

Control Fish (no blackcurrants):

- Normally I do not use such small snapper (700g) for my food preparations because the flesh is watery and not firm.

- I can tell immediately on slicing into the flesh to take the fillet that this fish is not fresh - there is no resistance to the knife.

The flesh does not have the desired consistency or appearance and is not suitable for sashimi. The skin is dark and not desirable.

I would not eat this fish.

I can tell by the appearance that this fish is very fresh. The skin colour is light and looks good.

The resistance of the knife on slicing in to take the fillet tells me this fish is very fresh, only a few hours old. The flesh has the right texture and translucence for snapper and is suitable for sashimi.

I will eat this fish. Further examination revealed that the weight of usable flesh from the trial fish was 30% greater than from the control fish after filleting.

Examination of the entrails revealed the trial fish had virtually no accumulation of fat around the gut whereas the control fish gut was coated with a fat layer. When the fish is not active, fat accumulates around the gut therefore the comparison indicates the trial fish had been more active in the pen through its life.

Exemplary formulation: Based on the above results, the inventors devised the following exemplary formulation.

A blackcurrant pomace is incorporated into pelleted, encapsulated or gelled fish feed. A blackcurrant pomace typically contains 50 - 300mg anthocyanin /100g pomace. The feed comprises:

Blackcurrant Pomace 80mg anthocyanin /100g pomace

Feed weight per fish per feeding 4.17g

Pomace incorporated into feed 180g per kg feed pellets produced

22.5kg into 125kg feed mix for pellets

This formulation provides a feed comprising 144mg anthocyanin /kg Feed.

The pomace and feed ingredients are mixed together and processed into the desired form such as, pellets, a gel, or capsules. It should be appreciated that the blackcurrant pomace may be substituted with other blackcurrant extracts or processed whole blackcurrants as desired to achieve a similar result. The exemplary formulation is particularly suited to snapper, or, for example, any other fish of a similar size and/or having a similar rate of growth. The amount of blackcurrant extract or processed whole blackcurrants combined in the feed may be varied to achieve the most cost effective outcome for a particular species. Example 2:

Further studies were completed to further analyse fish feeds of the invention.

The trials were conducted using fish feeds supplemented with blackcurrant in accordance with the invention. The feeds were in pellet, glazed pellet or flake form.

The pellet form comprised a dried blackcurrant pomace and standard fish feed ingredients including fishmeal, vegetable proteins, and binding agents such as wheat. The pellets were manufactured according to standard techniques. The dried blackcurrant pomace contained approximately 0.05 to 3.0% anthocyanin(s) and was present in the fish feed between approximately 0.1% to 5.0% of total fish feed. Pellet size varied depending on the size of the fish to be fed, according to well known principles.

The glazed pellet form comprised a food pellet comprising standard fish feed ingredients including vegetable protein, cereal grains, vitamins and minerals along with wheat to help bind the ingredients together. The pellets were manufactured according to standard techniques. The pellets were then glazed with a blackcurrant glaze (comprising a dried blackcurrant pomace powder, a dried blackcurrant puree powder, dried whole blackcurrant powder, dried blackcurrant concentrate powder, or blackcurrant concentrate) in accordance with standard glazing techniques. The glaze comprised approximately 0.05 to 3.0% anthocyanin(s) and comprised approximately 0.1% to 5.0% of the weight of total fish feed. Pellet size varied depending on the size of the fish to be fed, according to well known principles. Feed flakes comprised a blackcurrant component and standard fish feed ingredients including vegetable protein, cereal grains, vitamins, and minerals which were mixed together, then dried and made into small flakes according to standard techniques. The blackcurrant component contained approximately 0.05 to 3.0% anthocyanin(s) and was present in the fish feed between approximately 0.1% to 5.0% of total fish feed. Flake size varied depending on the size of the fish to be fed, according to well known principles.

Fish were feed the feeds to achieve a dose of between approximately 0.5mg to

approximately lOmg of anythocyanin/kg of body weight per dose.

The trials were conducted using a variety of different farmed fish species including:

prawns, salmon, shrimp and snapper.

After a period of feeding, fish were analysed against control fish (not feed the feeds of the invention) for a number of parameters. Results are provided below.

Brighter coloured muscles and flesh: Fish were cut open and it was observed that the muscle band is a lot brighter red in the fish species that were fed the feed of the invention compared to those which were not feed the feed of the invention. The flesh was brighter than the non blackcurrant fed fish.

Reduction in smell& improved taste: Fish were filleted. Fish feed with the feed of the invention had no farm odour compared to the fish which were given fish feed without blackcurrant inclusion. The taste of the fish fed the feed of the invention improved and was similar to that of fish being farmed in cleaner harvesting environments.

Oil Quality & fat content: There was an improvement in oil quality and a higher quality fat content in the fish which were fed the feed of the invention compared to the fish which were fed just the fish feed formulation without the blackcurrant inclusion.

Firmer Flesh: The flesh of farmed fish given feed according to the invention compared to those that weren't had firmer flesh with reduced gaping in the fillets. Weight of the flesh: The total weight of the usable flesh was up to 30% higher in the fish fed the feed of the invention than the fish that were not fed blackcurrant fish feed.

Shelf life extended: The shelf life of farmed fish fed the feed of the invention was extended for fresh fish markets by up to 48 hours longer than the non-blackcurrant fed fish and up to 72 hours longer for cooked fish markets compared to the non-blackcurrant fed fish.

Reliance of antibiotics: Most of the diseases fish get are through the gills. Typically, fish are farmed using antibiotics to control disease. Prawn, Shrimp, blue mackerel, paua, Ayu and Garden carp farms, in particular, use very high levels of antibiotics. These species are very vulnerable particularly because some are farmed at a young age (the death rate is high up to 1 year old) and others are mostly farmed commercially in fresh water. The inventors observed that regular feeding of the feed of the invention to baby prawns and Ayu resulted in a reduction of antibiotic use in commercial farms compared to those prawns and Ayu not fed the feed of the invention. Overall, the results show that death rate is reduced and the health of the aquatic animal is improved.

Reliance on astaxanthin supplementation: Astaxanthin is used as a feed supplement in many commercial fish farming environments. Its primary use is to provide a desired colour to the fish flesh. The level of astaxanthin used can vary from one geographical location to another and from one fish farm to another. In the location where the trials took place, the typical amount of astaxanthin is 30% to total feed weight. The inventors were able to reduce the astaxanthin levels by 50% through feeding the fish the feed of the invention. In addition, fish fed the feed of the invention were brighter than fish fed with standard fish feed and the colour of the flesh lasted longer in fresh and frozen fillets compared to fish fed feed without the blackcurrant component of the invention. Reducing the reliance on this compound has a number of benefits, including reduction in the cost of farming fish and reducing the reliance on synthetic compounds. Today, almost all commercial astaxanthin for aquaculture is produced synthetically from petrochemical sources. In addition, while it may constitute a small portion of fish feed, it typically makes up a major share of the cost, for example up to 20%. Often negative side effects present when aquatic animals are fed dietary supplements. In the trials conducted by the inventors there were surprisingly no negative side effects observed. The trials also indicated that the supplements and feeds of the present invention are palatable to various aquatic animals, as they readily consumed them. This was not expected, as aquatic animals find many compounds and composition unpalatable.

The results achieved indicate that the active ingredients in the supplements and feeds of the invention are readily bioavailable to the animals. There was no guarantee that this would be the case.

While blackcurrant compositions have been feed to land animals, a number of factors combined contributed considerable uncertainty as to whether such supplements could be delivered to, let alone effective in, aquatic animals. These include the facts that aquatic animals are very different in physiology to land animals, they have a different palate, they live in an aquatic environment and are often intensively farmed. The current invention now surprisingly shows that this can be achieved. The invention has been described herein, with reference to certain preferred embodiments, in order to enable the reader to practice the invention without undue experimentation. However, a person having ordinary skill in the art will readily recognise that many of the components and parameters may be varied or modified to a certain extent or substituted for known equivalents without departing from the scope of the invention. It should be appreciated that such modifications and equivalents are herein incorporated as if individually set forth. The invention also includes all of the steps, features, compositions and compounds referred to or indicated in this specification, individually or collectively, and any and all combinations of any two or more of said steps or features. Titles, headings, or the like are provided to enhance the reader's comprehension of this document, and should not be read as limiting the scope of the present invention. The entire disclosures of all applications, patents and publications, cited above and below, if any, are hereby incorporated by reference. However, the reference to any applications, patents and publications in this specification is not, and should not be taken as, an acknowledgment or any form of suggestion that they constitute valid prior art or form part of the common general knowledge in any country in the world.

Throughout this specification, unless the context requires otherwise, the words "comprise", "comprising" and the like, are to be construed in an inclusive sense as opposed to an exclusive sense, that is to say, in the sense of "including, but not limited to".