EP2890248A1 - Alleviation of anaemic growth suppression in fish - Google Patents
Alleviation of anaemic growth suppression in fishInfo
- Publication number
- EP2890248A1 EP2890248A1 EP13736528.4A EP13736528A EP2890248A1 EP 2890248 A1 EP2890248 A1 EP 2890248A1 EP 13736528 A EP13736528 A EP 13736528A EP 2890248 A1 EP2890248 A1 EP 2890248A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- fish
- heme
- vitamin
- source
- worm
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/20—Animal feeding-stuffs from material of animal origin
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/20—Animal feeding-stuffs from material of animal origin
- A23K10/22—Animal feeding-stuffs from material of animal origin from fish
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/20—Animal feeding-stuffs from material of animal origin
- A23K10/24—Animal feeding-stuffs from material of animal origin from blood
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/116—Heterocyclic compounds
- A23K20/132—Heterocyclic compounds containing only one nitrogen as hetero atom
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/174—Vitamins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/20—Inorganic substances, e.g. oligoelements
- A23K20/30—Oligoelements
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
- Y02A40/818—Alternative feeds for fish, e.g. in aquacultures
Definitions
- the present invention relates to the field of fisheries and aquaculture of all kinds, including the farming of fish, including saltwater or freshwater fish.
- the invention also relates to the treatment of fish to relieve conditions of anaemia.
- the invention concerns composition for fish food for alleviating anaemia in fish and increasing their growth rate Diminishing ocean fish stocks is causing a shift in fishing patterns and pressure in order to realise a goal of sustainable fisheries.
- Diminishing ocean fish stocks is causing a shift in fishing patterns and pressure in order to realise a goal of sustainable fisheries.
- that process of shift to sustainability is not guaranteed, nor can it guarantee to supply increasing human demand for fish.
- Fish farming and aquaculture provides an alternative to sustainable ocean fishing and the number of species of fish, crustacean and shellfish which are being farmed around the world is on the increase.
- Solea solea (the Dover sole) is a sandy and muddy bottom dwelling sea fish living down to about 300 metres that is exploited commercially by trawling.
- the diet of S. solea has been studied and found to include polychaete worms, small soft-shelled bivalves, small fishes and crustaceans.
- FEO UN Food and Agriculture Organization
- Dover sole has been found to grow well in aquaculture when it is fed on a diet of fresh Nereis virens (ragworm) and mussels, but this is not at present an economically viable way of producing the fish. Ragworm is expensive to produce. Conventional fish foods therefore continue to be used. As a result, the productivity of Dover sole in aquaculture systems continues to remain below expectations of what the fish is capable of in terms of growth. Much research therefore has been carried out to try and formulate an optimal diet for Dover sole in aquaculture, but without success. Various aspects have been investigated, including attractivity, nutritional composition, probiotic effects and processing effects. So far though, no one has succeeded in formulating an improved or optimal fish food for Dover sole. No currently available fish food composition or diet is able to realise the expected growth potential of Dover sole. The 25 - 65% faster growth rate of sole fed on ragworm compared to commercially available fish food shows that there is considerable growth potential to be achieved in sole and a fish food composition has yet to be formulated which would work to achieve such a faster growth rate.
- WO201 1/027279 A1 (UNIV DO ALGARVE et al) describes feed additives for aquaculture comprising 1 -methyl-L-tryptophan and/or its isomers.
- the compounds are disclosed as being olfactory attractants and inducers of digestion in fish and are for use at concentrations between 0.001 and 10 mg per kg of dry fish feed.
- the compounds were identified from seawater conditioned by the presence of 100g per liter of a polychaete worm Hediste diversicolor which was known to an important element of the natural diet of the Senegalese sole, S. senegalensis.
- the conditioned seawater was lyophilized to reduce its volume and then fractionated and tested on anaesthetised Senegalese sole subjected to olfactory receptor neurone electrode measurements. In the final fractionations mass spectrometry was used to identify 1 - methyl-L-tryptophan as the olfactory compound.
- the condition of anaemia is known to occur from time to time in various species of fish, salt or freshwater due to disease. Serious conditions are often manifest by pale gills and surface swimming and gulping.
- the disease agent may be a bacteria, a virus or a parasite, such as a leech or lice. For example, there is growing concern about incidences of anaemia in farmed salmon caused by the infections salmon anaemia virus (ISAV).
- CN101491669 A discloses a fish protein iron-peptide capsule (tablet or granule) for preventing iron deficiency anaemia, which is prepared by turning low-value marine fish and leftover protein into composite polypeptide through protein enzymatic hydrolysis, modifying the composite polypeptide with ferrous iron so as to obtain fish protein iron-peptide and mixing the fish protein iron- peptide with safflower seed oil, beeswax, phospholipids, starch according to a certain weight ratio.
- the invention discloses three formulations and one optimum
- the chemical nature of the capsule is alleged to be a chelated product of peptide, amino acid and ferrous iron.
- the protein content of the capsule is more than or equal to 30 percent, and the total iron content of each capsule is 10+/-0.5 mg.
- the capsule is a novel functional food for preventing iron deficiency anaemia.
- CN1644080 A (MINGBO AQUATIC PRODUCT CO LTD) discloses a feed for the half-smooth tongue-sole that is prepared from pulverized clam worm, oyster meat, fish powder, cybiidae powder, vitamin C, vitamin E, cod-liver oil, yeast, etc. Its advantages are easy absorption and digestion, and promoting growth.
- the inventors are surprised to discover from their work that by supplementing the diet of fish with annelid worm, an extract or fraction thereof, and/or a mollusc, mollusc extract or fraction thereof, that an anaemic state that holds back growth of the fish is reduced or overcome.
- the "scope for growth" of the fish is made accessible and the inventors refer to this as the "the worm effect”.
- the inventors have also surprisingly discovered that the "worm effect” is due to heme. Also discovered is that the "worm effect” includes a combination of heme and vitamin Bi 2 . Further, the inventors have discovered that best increases in fish growth rate compared to available commercial feeds arise when there is a combination of added heme and vitamin B 2 in the diet and there is a level of taurine presence comparable to the levels generally found in fishmeal .
- the present invention provides a method of increasing the productivity of a system for culturing fish comprising feeding the fish a diet, which diet comprises a source of heme.
- a diet which diet comprises a source of heme.
- SGR specific growth rate
- %.d “1 , growth in metabolic body weight (g.kg “ 0 8 .d “1 ) and/or growth in gram per day is realised compared to non-supplemented feeds.
- Any source of heme is suitable so long as iron is bound to heme and not free.
- bovine haemoglobin, iron sulphate hydrate, iron proteinate, or iron methionate for example, bovine haemoglobin, iron sulphate hydrate, iron proteinate, or iron methionate.
- Productivity of the system of the invention may be as much as 287% of ordinary pellet fed farmed fish; optionally 243%, 197% 167% or 135%.
- the diet may further comprise a source of vitamin Bi 2 .
- Vitamin B12 or cobalamin belongs to a group of cobalt containing compounds known as corrinoids that contain a specific corrinring. Known forms are methylcobalamin, adenosylcobalamin, hydroxocobalamin, cyanocobalamin, sulfitocobalannin and possibly other unknown forms.
- source of heme is heme itself, and within the term “source of vitamin B 2 " is vitamin B 2 itself.
- a culture system for fish includes all kinds of aquaculture and fish farming, including "sea ranching" whereby fish are allowed to roam but they are preconditioned to go to a feeding area at specific times where they are rewarded with the free availability of food which may include molluscs and/or annelids.
- the diet of the invention preferably has a taurine level comparable to levels found in fish meal. Particularly advantageous increases in growth rate of fish are found where there is a combination of heme and vitamin Bi 2 and a level of taurine comparable to levels found in fishmeal or compared to commercially available fish diets based on fish meal.
- the diets of the invention contain taurine in the range 3.1 to 7.6. kg "1 .dm.
- 2 optionally vitamin B-
- the source of heme may be an annelid worm or an extract or fraction thereof;
- the annelid worm is a member of the class Polychaeta; preferably a ragworm or is a member of the family Nereidae; more preferably wherein the annelid worm is of the genus Nereis; even more preferably the species Nereis virens.
- the source of heme may be a mollusc or an extract or fraction thereof; optionally wherein the mollusc is a species of the phylum Mollusca; preferably the class Bivalvia.
- the source of heme is not an annelid worm or an extract or fraction thereof; and not any annelid worm being is a member of the class Polychaeta; not a ragworm or a member of the family Nereidae; also not an annelid worm of the genus Nereis; and not species Nereis virens.
- the source of heme is not a mollusc or an extract or fraction thereof; and not a species of the phylum Mollusca; and not a member of class Bivalvia.
- the source of vitamin Bi 2 may be fermented salmon kidneys.
- a known preparation called “Mefun” which is a natural product with the highest known Bi 2 content (about 3280 ⁇ g.kg "1 ).
- the heme and vitamin B 2 may be mixed with and thereby may form part of the food given to the fish.
- this allows easier supplementation and feeding regimes in that fish may be fed from a single bulk supply of pre-prepared food.
- 2 may be formulated to be given to the fish alongside the food as a separate supplement, but not actually mixed into the normal food to form part of it.
- the heme and vitamin B 2 may be formulated as a mixture or as separate additives.
- the fish is preferably a saltwater fish, although a freshwater fish may be the subject of the method of the invention.
- a preferred fish subjected to the method of the invention may be a member of the family selected from Solidae, Achiridae or Cynoglossidae.
- An alternatively preferred freshwater fish is selected from the family Cyprinidae; preferably Danio rerio (zebra fish).
- preferred fish are Solea solea or Solea senegalensis.
- such fish may be selected from: Seriola quinquieradiata (Japanese amberjack), Epiniphelus fusoguttatus (Tiger grouper), Onchrhynchus kisutch (Coho salmon), Sparus aurata (Giltehead sea bream) Pagrus major (Red sea bream), Gadus morhua (cod), Sander lucioperca (Pike), Oncorhynchus mykiss (Rainbow trout), Salvelinus alpines (Arctic char), Oncorhynchus tshawatcha
- Dipolodus sargus White sea bream
- Oreochromis niloticus Ne tilapia
- Pseudosciaena crocea Large yellow croaker
- Pollachius pollachius Policy Pollachius
- Dicentrarchus labrix Sea bass
- Hippoglossus hippoglossus Halibut
- Lates calcarifer Barramundi
- Morone saxatilis String bass
- Coryphaena hippurus Mahi mahi
- Pagellus bogaraveo Black spot sea bream
- Paralichtys olivaceus Bastard halibut
- Cyprinus carpio Carp
- Scopthalmus/Psetta maximus Turbot
- Salmo salmar Almar
- Epinephelus coioies Green grouper
- Anguilla anguilla European eel
- the annelid worm, extract or fraction, and/or the mollusc, extract or fraction is preferably in the form of a meal and/or is freeze dried. Where the annelid worm or mollusc is taken and used as the whole animal, then it would usually be dried and processed to form a meal, but without boiling. Alternatively, the whole animals may be freeze dried and then physically processed to form flakes, granules or powder.
- the average temperature of the water in which the fish are maintained is depending on the species and size of species cultured. For common sole (Solea solea) during grow-out this is preferably not less than about 15.5 °C; more preferably the average temperature is a temperature falling in the range 15.5 °C to 20 °C; even more preferably an average temperature of 17.5 °C. Growth of fish at such average temperature regimes using the diets of the present invention advantageously results in a reduction or elimination of nutritionally incurred anaemia and optimally increased growth rate compared to a normal, non-supplemented diet. Similar temperature ranges apply for other species.
- Fish grown in accordance with the invention may have a growth rate (in g per day) which is at least 5% or at least 10%, or at least 15%, or at least 20%, or at least 25%, or at least 30%, or at least 35%, or at least 40%, or at least 45%, or at least 50%, or at least 55%, or at least 60%, or at least 65%, or at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 90%, or at least 95%, or at least 100% greater than the equivalent growth stage of fish fed the same fish food but lacking the essential source of heme and source of vitamin B-
- a growth rate in g per day
- a culture system for fish may comprises a body of marine water and associated marine bed; optionally delineated at least in part by a physical barrier, e.g.
- the culture system may comprise a tank of seawater.
- the invention also provides heme for use as an oral medicament in fish.
- the invention further provides heme in combination with vitamin B 2 for use as an oral medicament for fish.
- the nature of the oral medicament is such that it is in any form ingestible by fish, preferably by self-feeding from the medicament being made available in the form of a food or a supplement provided with food.
- the invention includes a combination of heme and vitamin Bi 2 for use in the prevention or treatment of anaemia in fish, wherein the heme and vitamin B 2 are in the form of an oral composition.
- the invention includes a method of preventing or treating anaemia in fish comprising providing a combination of heme and vitamin B 2 to the fish.
- the invention also includes a method of increasing haematocrit in a fish comprising feeding the fish with a fish food supplemented with heme and vitamin B-
- such medicaments and compositions are preferably administered as a food supplement or included within food forming the general diet of the fish.
- the optional and preferred features of the method of the invention as hereinbefore defined apply equally to the aforementioned medical uses of the invention.
- the haematocrit of the treated fish is at least about 16%; preferably the haematocrit is in the range 16 - 30 %; more preferably 16 - 20%.
- the haematocrit of treated fish may therefore be 17% + 1 %; or 18%; + 1 %; or 19 + 1 %; or 20% + 1 %; or 21 % + 1 %; or 22%; + 1 %; or 23% + 1 %; or 24% + 1 %; or 25%; ⁇ 1 %; or 26% + 1 %; or 27% + 1 %; or 28% + 1 %; or 29%; + 1 %; or 30% + 1 %. Whilst specific haematocrit levels may differ in other fish species, values for sole may be considered broadly representative in many other species.
- the haemoglobin level of the treated fish is at least about 22 g/l; preferably the haemoglobin is in the range 22 - 47 g/l; more preferably in the range 22 - 27 g/l.
- the haemoglobin level of the treated fish may therefore be 22 + 1 g/l; or 23 + 1 g/l; 24 + 1 g/l; or 25 + 1 g/l; 26 + 1 g/l; or 27 + 1 g/l; 28 + 1 g/l; or 29 ⁇ 1 g/l; or 30 ⁇ 1 g/l; or 31 + 1 g/l; 32 + 1 g/l; or 33 ⁇ 1 g/l; 34 + 1 g/l; or 35 + 1 g/l; 36 + 1 g/l; or 37 + 1 g/l; 38 + 1 g/l; or 39 + 1 g/l; or 40 + 1 g/l; or 41 + 1 g/l; 42 + 1 g/l; or 43 + 1 g/l; 44 + 1 g/l; or 45 + 1 g/l; 46 + 1 g/l; or 47 + 1 g/l. Whilst specific haemoglobin levels may differ in
- the specific growth rate (SGR) of the treated fish is greater than about 0.75 % body weight per day (%bw.d "1 ).
- the SGR of treated fish is at least about 0.8 %bw.d “1 ; or at least 0.9 %bw.d “1 ; or at least 1 .0 %bw.d “1 ; 1 .1 %bw.d “1 ; or at least 1 .2 %bw.d “1 ; or at least 1 .25 %bw.d “1 ; or at least 1 .3 %bw.d “1 ; or at least 1 .4 %bw.d “1 ; or at least 1 .5 %bw.d "1 ; or at least 1 .6 %bw.d “1 ; or at least 1 .75 %bw.d “1 .
- the growth rate of fish may be greater than about 0.8 g per day (g.d "1 ).
- Preferred growth rates of fish may be greater than about 0.9 g.d “1 ; or greater than about 1 .0 g.d “1 ; or greater than about 1 .0 g.d “1 ; or greater than about 1 .1 g.d “1 ; or greater than about 0.9 g.d “1 ; or greater than about 1 .0 g.d “1 ; or greater than about 1 .25 g.d "1 .
- Fish which may be subject to the various aspect of the invention may be at any stage from juvenile (e.g. from as early as weaning from yolk sac to artificial feeds) to adult. Fish as small as 0.03 g may be subjected to the methods of the invention. Mature fish such as Sole, grown or treated in accordance of the invention, may have a weight in the range 50g to 350g, for example.
- the invention provides a method of making fish food composed of a proteinaceous material of animal and/or fish and/or vegetable origin, comprising adding a source of heme to the proteinaceous material.
- the method of making fish food further comprises adding a source of vitamin Bi 2 .
- the source of heme and optionally the source of vitamin B 2 may be as set forth in relation to the method of increasing productivity invention as hereinbefore defined.
- the source of heme and optionally vitamin Bi 2 may be incorporated into fish food pellets by a process of cold extrusion with the fish food composition.
- a conventional fish food may be supplemented with a source of heme and optionally a source of vitamin Bi 2 .
- 2 may be as set forth in hereinbefore defined.
- the conventional fish food may be made by grinding and mixing together ingredients such as fishmeal, fish oil, vegetable proteins and binding agents, e.g. wheat.
- the resultant past water may be added to improve consistency
- the diameter of the pellets depending on the species and size (growth stage) of species cultured.
- the time of adding oil is depends on the process used and species specific requirements (fat requirements, or e.g. the need for floating, slow sinking or sinking pellets). In the process producing pellets e.g. for Sole, (a low fat diet) the oil is mixed with the other ingredients before forming.
- the temperature at which pellets are formed and the amount of oil affects their density and therefore whether they float or sink.
- the supplemental ingredients of the invention may be added before, during or after pelleting.
- Commercial fish foods are readily obtainable from the likes of Biomar, EWOS or Skretting.
- a method of feeding fish in accordance with the invention comprises making a proteinaceous fish food available to the fish and whether prior to, separately, simultaneously or subsequent to making food available, administering a source of heme; optionally also a source of vitamin B 2 .
- kits for improving the productivity of an aquaculture system comprising at least one container which contains a source of heme; optionally wherein a source of vitamin B 2 is provided in the same or different container to the source of heme.
- the at least one container is graduated, or the at least one container is a unit dose of heme, optionally a unit dose of vitamin B 2 in addition.
- the kit may include a set of instructions for generating the correct level of supplement to the conventional fish food.
- the invention also includes a fish food comprising protein of plant and/or animal origin, gluten, a binder, salt, minerals and vitamins, and supplemented with a source of heme; optionally also a source of vitamin Bi 2 .
- a level of taurin comparable to levels found in fish meal.
- taurin levels vary between 3.1 and 7.6 g.kg "1 .dm.
- the supplement may be present in an amount in the range 5 - 85% by weight of the total food; preferably an amount in the range 10 - 75% by weight.
- Other preferred ranges include 15 - 65%, 20 - 60%, 25 - 55%, 30 - 50% or 35 - 45%.
- the supplement may be present in an amount in the range 5 - 10%; 10 - 20%; 20 - 30%, 30 - 40%, 40 - 50%, 50 - 60%, 60 - 70% or 70 - 80%.
- the supplement may be present in an amount of about 10%, or about 25%, or about 50%, or about 75% by weight of total food. .
- the plant protein may be pea protein and/or soy protein, e.g. soy protein
- the animal protein may be casein.
- Fish foods of the invention may also comprise one or more of the following
- an oil e.g. a fish oil
- sugar e.g. a fish oil
- sugar e.g. a fish oil
- wheat gluten e.g.
- salt choline chloride
- vitamin C e.g. a fish oil
- vitamin E e.g. a fish oil
- preservative e.g. BHT
- antifungal e.g. calproprionate
- the source of heme and optionally source of vitamin B-12 is preferably not heat treated in excess of about 55°C, and in particular is not subjected to boiling for any period of time.
- Fish food which is supplemented in accordance with any aspect of the invention may have an amount of one or more of the following added to it to increase concentration thereof, whether prior to, during or after pellet formation: e.g. vitamin C, folic acid, vitamin B1 , vitamin B2, vitamin B5, vitamin A, iron, zinc, selenium, and copper etc. (e.g. see premix table 7).
- Figure 1 shows levels of oxygen, pH and temperature as monitored during the trial of example 1 investigating growth of sole fed on diets of fresh worm, pellets of a commercial fish food with extract of worm and untreated pellets of a commercial fish food.
- Figure 2a shows the average feed intake in g. dm. fish "1 .d “1 as described in example 1 .
- Figure 2b shows the average growth in g.d "1 and standard deviation of fish fed ragworm, pellet treated with worm extract and the commercial pellet, as described in example 1 .
- Figure 3 shows average haematocrit values for the sample group t(0) , negative control and pellets treated with extract do not differ, as described in example 1 .
- FIG. 4 shows the relationship found in example 1 between feed intake d m and temperature °C of culture water for sole fed pellet, pellet with extract and fresh worm.
- Figure 5 shows the relationship found in example 1 between temperature °C, htc and oxygen use (in ml.min “1 .kg "1 ) calculated from the feed intake per tank (following pattern of figure 4). Real oxygen use is expected even lower as requirements for maintenance (basic metabolic oxygen demand) are not included.
- Figure 6 shows the growth in metabolic bodyweight of fish fed ragworm from example 1 ; the treated and untreated pellet in period 1 , period 2 and the average of the total trial.
- Figure 7 shows the growth in metabolic bodyweight of fish fed ragworm from example 1 ; the treated and untreated pellet in period 1 , period 2 and the average of the total trial.
- Figure 8 shows the relationship between haematocrit (%) and growth in metabolic body weight in g.kg “0 8 .d "1
- Figure 9 shows the result from example 1 of feeding boiled worm compared to fresh worm has a negative effect on growth of sole, especially in period 1 with the higher water temperatures.
- Figure 10 shows the result from example 1 that feeding boiled worm compared to fresh worm has a negative effect on haematocrit level of sole, although still higher than fish fed the commercial pellet.
- Figure 1 1 shows from example 2 the relationship between worm meal inclusion level and growth in metabolic body weight.
- the inclusion level of 100% worm meal is the fresh worm.
- Figure 12 shows from example 2 the relationship between worm meal inclusion level and haematocrit (%).
- the inclusion level of 100% worm meal is the fresh worm.
- Average haematocrit levels measured in fish fed the commercial pellet was 1 1 .2%.
- Figure 13 is from example 2 and shows the relationship between haematocrit and growth.
- Figure 14 shows the relationship between iron content and growth in metabolic body weight. Iron content of pc (worm), nc (Weanex) and worm meal were analysed. Iron content of diets G, B, C, D and E were calculated.
- Figure 15 shows the relationship between the calculated heme content, haematocrit and growth in metabolic body weight. Heme content of fresh worm analysed was comparable as values calculated using the data of Vinogradoff et al. (1991 ) "Iron and heme contents of the extracellular haemoglobins and chlorocruorins of Annelids.” Comp. Biochem. Physiol. Vol. 98B, No. 2/3, pp. 187-194.
- Figure 16 is from example 3 and shows the recovery pattern of sole fed ragworm (N. virens).
- Figure 17 shows the relationship between haematocrit and haemoglobin as measured in the trial of example 3.
- Figure 18 shows the average haematocrit level in percentage (left) and the average haemoglobin levels in g.l "1 (right) and the standard deviations of sole fed ragworm, mussel and the commercial pellet.
- Figure 18 shows the relation between bodyweight in gram and the specific growth rate (SGR) in %.d "1 of sole fed live ragworm.
- Figure 19 shows the weight gain (g, in time (days) of sole fed live ragworm.
- Fermented salmon kidneys or "Mefun” is a natural product with the highest known B 2 content (3280 ug.kg “1 ). "Mefun” can therefore be substituted for RMM (raw mussel meat) and/or ragworm, with equivalent or greater effect of alleviating Sole from anaemic growth suppression.
- sole Under commercial conditions using recirculation systems (RAS) and available commercial feeds sole will grow from 50 to 150 g in ⁇ 9 months. It will take sole fed commercial pellets ⁇ 21 months or more than twice as long compared to sole fed worm to grow to sizes of ⁇ 350 grams.
- RAS recirculation systems
- Figure 19 shows the combined growth curve using two different size classes: a group that grew from 50 to 235 grams in 150 days and a group that grew from 210 to 370 gram in 150 days. From this figure one can extrapolate that sole fed ragworm can grow from 50 to 350 grams in 275 days.
- the pellet of the invention achieved growth figures of 0.73%.d “1 , 4.96 g.kg 0 8 .d “1 and 1 .06 g.d “1 for SGR, Growth in metabolic body weight and growth in gram per day using sole of around 150 grams.
- this experiment there was no reference included in terms of a commercial pellet or worm, but when comparing this figure with the known commercial data the results are better.
- Example 1 Identification/screening of intrinsic factors of the ragworm (Nauss virens) responsible for the excellent growth of sole (Solea solea)
- the experimental set-up contained five diets: (1 ) freshly chopped ragworm, (2) boiled chopped ragworm, (3)
- frozen/thawed worm (4) a pellet flavoured with worm extract, and (5) a commercial pellet as a negative control
- Weanex, 3mnn, Biomar. Diets were tested in triplicate, except fresh worm treatment, which was tested in 4 tanks, giving 16 experimental units, 15 fish per tank, for a period of 64 - 67 days due to sampling of one block per day. Tanks within treatments were divided over 4 different blocks. Boiled worms were cooked for three minutes before being chopped. Frozen worms were stored for 24 hours at -20°C and thawed before being fed.
- Extracts for flavouring pellets were produced by homogenizing cooled ragworms (Topsy Baits B.V.) using a blender. Homogenates were centrifuged (20 min,
- Feeding Fish were fed by hand twice a day (8:30 and 16:30) and amount was recorded accordingly. Feeding level was equal for all tanks and adapted daily towards the highest feeding level possible. During acclimatization (19 days) fish were fed the commercial diet. Worms were chopped using a knife and chopping board and sieved for 1 min to drain excess fluids prior to weighing.
- Feed intake and feed conversion Feed intake in g.d "1 , as sum of feed given in the morning and afternoon, was estimated by number of pellets given minus number of pellets recovered, multiplied by the average pellet weight or weight worms given, minus the weight worms recovered. As dry matter content varied between
- Feed conversion on dry matter was calculated applying the following formula 1 :
- FCRDM (Fl * dry matter experimental diet)/growth.
- Worms were sampled daily. Each day adequate quantities of ragworm were taken and stored at -20°C for further analysis. Quantities were based on the average feed intake per treatment from the previous day. Daily samples were pooled per treatment after the end of the experiment to get the average dry matter content and proximate composition.
- Blood 10 fish at start and 5 fish of every tank at the end of the experiment were sacrificed to collect blood. Blood was obtained by caudal venous puncture using a heparinized syringe (0.6mm/30mm needle) as soon as fish was anaesthetized. After collection, blood was immediately transferred into labelled tubes and stored on ice until further processing.
- haematocrit A little amount was used for determination of haematocrit by using heparinized tubes (0 1 .5mm, L 75mm) and a haematocrit centrifuge (standard rpm during 5 minutes). The remaining part of the sample was centrifuged at (10000 x g, 4°C, 10 min) to collect plasma. Plasma was transferred to clean tubes and stored at -80°C until further analysis.
- Statistical analysis Data were analysed using ANOVA to test for block and treatment effects. The model assumes that the residual effects (e) are
- Feed intake and growth at day 31 for worm tank 1 , 2, 3 and 4 were 0.700, 0.738, 0.755 and 0.466 g dm.fish “1 .d “1 and 6.16, 6.58, 5.64 and 2.43 g.kg “0 8 .d “1 , respectively.
- the experimental units were completely independent (flow through) and both feed intake and growth are strong indicators of welfare and health of fish, it was clear that there was something wrong with the latter population and it was decided to discard this tank from the experiment. Mortality during the experimental period in the other tanks was limited to three fish and was not related to treatments.
- Average values for temperature, oxygen, pH, TAN, NO 2 , NO 3 , and flow.tank "1 during the trial were 15.7 , 8.9mg.l “1 , 8.16, 0.07 mg.l “1 , 0.05 mg.l “1 , 1 .14 mg.l “1 , 2.5 l.min “1 , respectively.
- the average temperature for period 1 and period 2 were 17 ° C and 14 ° C respectively.
- the daily temperature and oxygen levels during the experiment and divided over period 1 and 2 are shown figure 1 . Dry matter content of diets were 89.1 %, 89.6% and 18.0% for the untreated pellet, treated pellet and worm respectively.
- P values from F test, testing for block effects show no significant block effects.
- Feed intake and growth between fresh worm, pellet with extract and untreated pellet Feed intake on dry matter base does significantly differ between diets
- Table 1 below shows body weight at start (BW s t ar t), body weight at end (BW end ), growth, SGR, growth in metabolic body weight, hepatosomatic index (HSI), feed conversion on dry matter base (FCR dm ) of the different treatments.
- Haematocrit and hepatosomatic index For haematocrit, the effect of
- HSI hepatosomatic index
- HAI Hepatosomatic Index
- Htc Hepatosomatic Index
- Htc Hepatosomatic Index
- g.dm Feed intake g.dm '1 .fish '1 .cT 1
- Haematocrit, feed intake and growth in relation to temperature The current trial can be divided in two periods (see figure 1 ). Period P1 has an average temperature of 17.2 °C and P2 has an average temperature of 14.5 °C. Due to these different periods relations between haematocrit, temperature, feed intake and growth could be made which are shown and briefly discussed below.
- Feed intake in relation to temperature Use of worm extract equalizes the
- haematocrit The relationship between haematocrit and growth is shown in figure 8. What is clear is that low haematocrit levels (anaemia) in fish compromises O2 transport and metabolic performance. Therefore it is likely that the need for O 2 plays a pivotal role in setting the lower limit for haematocrit in normocythemic fish. Virtually nothing is known about the effects of physiological and environmental influences on haematocrit regulation, or the links between haematocrit, haemoglobin, nutrition, and growth (Gallaugher and Farrell (1998)).
- haematocrit anaemia
- the anaemia observed in sole fed commercial pellet is a form of nutritional anaemia.
- Temperature is critical at about 16 °C.
- Worm increases haematocrit and thereby "scope for growth" at higher temperatures.
- Boiling has a negative effect on the properties of fresh worm as shown by lower growth and lower haematocrit of fish fed with boiled worm compared to fish fed fresh worm.
- Example 2 The "worm factor”; a strong dose response relation between worm meal inclusion level, haematocrit and the growth of sole (Solea solea).
- Fresh worm has no added value.
- Experimental design, diets and preparation The experimental set-up contained seven diets; a diet with 0, 10, 15, 50 or 75% of worm meal, fresh worm as a positive control, which is referred to as the diet with 100% worm meal inclusion level, and a negative control (Weanex3mm, Biomar). Diets were tested in triplicate, giving 21 experimental units, 15 fish.tank "1 , for a period of 42 days for growth and 45, 46 and 47 days for other parameters due to the necessity of sampling of one block per day. Ragworms were provided three times a week by Topsy Baits B.V. The experimental recipes are shown in table 3. Diets were prepared by cold extrusion.
- RVET (g/kg) 134.8 134.8 134.8 134.8 134.8
- Composition diet B, C, D, E and G are equal in macronutrients, amino acids, calcium and phosphates
- Table 4 the vitamin and mineral premix of Borges et al (2009): Vitamin and mineral premix
- Vitannine A (retinyl acetate) 2.4 IU)
- Environmental parameters for photoperiod, light intensity, temperature, O 2 , pH and salinity were 12L:12D, low;1 1 -15 lux, ⁇ 18°C, 6-8mg.l "1 , 7.5- 8.0, and 25-30ppt, respectively. Temperature and O 2 were measured daily. Flow, pH, TAN, NO2 were measured weekly. Mortality, date and weight of dead animals were recorded.
- Feeding Fish were fed by hand twice a day (8:30 and 16:30). Feeding level was equal for all tanks and adapted daily towards the highest feeding level possible. After an hour uneaten feed was removed from tanks randomly after every feeding period. During acclimatization (14 days) fish were fed the commercial diet. Worms were chopped using a knife and chopping board and sieved for 1 min to drain excess fluids prior to weighing.
- Worms were sampled daily. Each day adequate quantities of ragworm were taken and stored at -20°C for further analysis. Quantities were based on the average feed intake per treatment from the previous day. Daily samples were pooled per treatment after the end of the experiment to get the average dry matter content and proximate composition.
- Blood 10 fish at start and 5 fish of every tank at the end of the experiment were sacrificed to collect blood. Blood was obtained by caudal venous puncture using a heparinized syringe (0.6mm/30mm needle) as soon as fish was anaesthetized. After collection, blood was immediately transferred into labelled tubes and stored on ice until further processing. A little amount was used for determination of haematocrit by using heparinized tubes (0 1 ,5mm, L 75mm) and a haematocrit centrifuge (standard rpm during 5 minutes). The remaining part of the sample was centrifuged at (10000 x g, 4°C, 10 min) to collect plasma. Plasma was transferred to clean tubes and stored at -80°C until further analysis.
- Example 3 Sole fed ragworm recover from their anaemia in 15 - 21 days
- the experiment consists of 13 different treatments, 10 fish.tank "1 ; of which12 with fish fed freshly chopped ragworm using a regression set up of which the sample times were spread over 26 days (day 0, 2, 4, 6, 8, 10, 13, 16, 19, 21 , 23 and 26) and one with fish fed a commercial diet (Pelleted feed, 3mm, from a regular producer) as a control group which got sampled at day 26.
- Treatments were allocated randomly over available tanks. Ragworms were provided three times a week by Topsy Baits B.V.
- Environmental parameters for photoperiod, light intensity, temperature, O 2 , pH and salinity were 12L:12D, low;1 1 -15 lux, ⁇ 18°C, 6-8mg.l "1 , 7.5 - 8.0, and 25-30ppt, respectively. Temperature and O2 were measured daily. Flow, pH, TAN, NO 2 " were measured weekly. Mortality, date and weight of dead animals were recorded.
- Feeding Fish were fed by hand twice a day (8:30 and 16:30) and uneaten feed was removed randomly after an hour. Feeding level was equal for all tanks and adapted daily towards the highest feeding level possible. During acclimatization (9 days) fish were fed the commercial diet. Worms were chopped using a knife and chopping board and sieved for 1 min to drain excess fluids prior to weighing.
- Blood 20 fish at start and 10 fish of every tank were sacrificed to collect Blood. Blood was obtained by caudal venous puncture using a heparinized syringe
- Haematocrit was determined by using capillaries and a haematocrit centrifuge (standard rpm during 5 minutes). Haemoglobin content of blood was determined using the method described by Kampen and Zijlstra (1961 )
- Example 4 The effect of mussel (Mytilus edulis) is comparable to the effect of ragworm (Nauss virens) on alleviation of anaemic growth suppression in Dover sole (Solea solea)
- Htc levels of sole fed worm or RMM are not significantly different.
- the Hb levels of sole fed pellets (18.92 g.l “1 ) is significantly lower compared to the Hb level of sole fed RMM (26.70 g.l “1 ) or worm (33.72 g.l “1 ).
- the Hb level of sole fed worm is higher than that of sole fed RMM.
- Feeding sole RMM does increase Htc and Hb to comparable levels as with ragworm.
- mussel and ragworm are considered substantially equivalent in the alleviation of anaemic growth suppression in Dover sole.
- the experimental set-up contained three diets; raw mussel meat (RMM), fresh worm and a commercial diet (extruded, 3mm, from a regular producer). The diets were tested in triplicate, giving 9 experimental units with 15 fish.tank "1 for a period of 23 days. Mussels were collected at the Oosterschelde, the Netherlands. The RMM was separated from their shell, all at the same day, and frozen at -80°C until needed for feeding. The frozen mussel meat was thawed slowly in a fridge the day before feeding. Ragworms were provided three times a week by Topsy Baits B.V, The Netherlands A 7-day acclimatization and a 23-day experimental period was used for Sole of weight ⁇ 220g.
- the fish were distributed within 9 tanks (0.4m 2 , 1301) having integrated flow through system, using sand filtered seawater with a flow of 4 l.min " 1 .tank "1 .
- fish were preventively treated with formalin (O.lg.l “1 , duration 12 hours).
- Environmental parameters for photoperiod, light intensity, temperature, O 2 , pH and salinity were 12L:12D, 1 1 -15 lux, ⁇ 18°C, 6-8mg.l "1 , 7.5-8.0, and 25-30ppt, respectively.
- Temperature and O2 were measured daily.
- Flow, pH, TAN, NO 2 " were measured weekly. Mortality, date and weight of dead animals were recorded.
- Example 1 The feeding of fish, diets, sampling, analysis, measurements, blood measurements and calculations were carried out substantially as described in Example 1 .
- the average water quality parameters for oxygen, temperature, pH, ammonia, nitrite and nitrate were 16.9, 8.3, 8.1 , 0.3, 0.0, 4.5 respectively and stayed within range.
- the hepatosomatic index (HSI), the haematocrit (Htc) and haemoglobin (Hb) values at the start of the experiment were 1 .13%, 12.48% and 19.46 g.l "1 , respectively.
- the experimental set-up contained nine diets (see table 6); 4 diets with haemoglobin as iron source, a diet with Fe(SO ) as an inorganic iron source, a diet with iron proteinate (JH Biotech, Inc), a diet with iron chelate (JH Biotech, Inc), a positive control (worm, N. Virens) and a commercial pellet. Diets enriched with haemoglobin were each different in a combination of high vs. low level of vitamin B 2 and taurine (table 4). Two levels of taurine are included in the experiment as taurine is essential for efficient nutrient digestion.
- Diets were tested in triplicate, giving 27 experimental units, 15 fish.tank "1 , for a period of 23 days. Diets were prepared using cold extrusion. Table 7 shows the content of the diets.
- Composition of diet B, C, D, E, F, G and H are equal in macronutrients, amino acids, calcium and phosphates.
- Table 7 Diet formulations, inclusion level of different test products and calculated composition.
- Binder 1 p 2.00 2.00 2.00 2.00 2.00 2.00 2.00 2.00 2.00 2.00
- Binder 2 q 2.00 2.00 2.00 2.00 2.00 2.00 2.00 2.00 2.00
- Vitamin Bi 2 2 0.190 0.190 0.028 0.190 0.028 0.190 0.190
- RVET (g.kg "1 ) 108 108 108 108 108 108 108 108 108 108 108 108 108 108 108 108 108 108 108 108 108
- RVET (g.kg “1 ) 130.0 122.0 125.0 123.0 125.0 130.0 127.0
- Vitamins (mg or IU kg-1 diet): vitamin A (retinyl acetate); 2.7 mg, 9000 IU; vitamin D3 (cholecalciferol), 0.04 mg 1700 IU; vitamin K3 ((menadione sodium bisulfite), 10 mg; vitamin B1 (thiamine), 10 mg; vitamin B2 (riboflavin), 20 mg; vitamin B6
- Htc haematocrit
- Hb haemoglobin
- HIS hepatosomatic index
- Vitamin B 2 has a positive effect on haematocrit in sole fed diets formulated with heme. There is an interaction between between B 2 and taurine whereby high levels of taurine reduce the anaemia alleviating effects. Therefore a combination of heme with high B-
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CN101491669A (en) | 2008-08-04 | 2009-07-29 | 浙江海洋学院 | Anti-anaemia fish-protein iron peptide capsule |
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Non-Patent Citations (6)
Title |
---|
CADENA ROA M ET AL: "USE OF REHYDRATABLE EXTRUDED PELLETS AND ATTRACTIVE SUBSTANCES FOR THE WEANING OF SOLE (SOLEA VULGARIS)", JOURNAL OF THE MARICULTURE SOCIETY, LOUISIANA STATE UNIVERSITY, BATON ROUGE, LA, US, vol. 13, 1 January 1982 (1982-01-01), pages 246 - 253, XP008029006, ISSN: 0735-0147 * |
JEROEN KALS ET AL: "Mussel ( Mytilus edulis L.) and ragworm ( Nereis virens , Sars) both alleviate anaemia in common sole ( Solea solea L.)", AQUACULTURE RESEARCH, 8 August 2015 (2015-08-08), GB, pages n/a - n/a, XP055278024, ISSN: 1355-557X, DOI: 10.1111/are.12871 * |
MARK J. KOURY ET AL: "NEW INSIGHTS INTO ERYTHROPOIESIS: The Roles of Folate, Vitamin B 12 , and Iron* 1", ANNUAL REVIEW OF NUTRITION., vol. 24, no. 1, 14 July 2004 (2004-07-14), US, pages 105 - 131, XP055277753, ISSN: 0199-9885, DOI: 10.1146/annurev.nutr.24.012003.132306 * |
R MÉTAILLER ET AL: "ATTRACTIVE CHEMICAL SUBSTANCES FOR THE WEANING OF DOVER SOLE (Solea vulgaris): QUALITATIVE AND QUANTITATIVE APPROACH", JOURNAL OF WORLD MARICULTURE SOCIETY, 1 March 1983 (1983-03-01), pages 679 - 684, XP055352416, Retrieved from the Internet <URL:http://onlinelibrary.wiley.com/doi/10.1111/j.1749-7345.1983.tb00121.x/abstract> [retrieved on 20170307] * |
S KROECKEL ET AL: "Comparing feed intake, utilization of protein and energy for growth and body composition in S. solea fed natural and commercial diets", 1 January 2009 (2009-01-01), pages 1 - 23, XP055082531, Retrieved from the Internet <URL:http://edepot.wur.nl/143313> [retrieved on 20131004] * |
See also references of WO2014032837A1 * |
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