CN102469808A - Use of natural substances as feed additives for aquatic animals - Google Patents
Use of natural substances as feed additives for aquatic animals Download PDFInfo
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- CN102469808A CN102469808A CN2010800323879A CN201080032387A CN102469808A CN 102469808 A CN102469808 A CN 102469808A CN 2010800323879 A CN2010800323879 A CN 2010800323879A CN 201080032387 A CN201080032387 A CN 201080032387A CN 102469808 A CN102469808 A CN 102469808A
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- eugenol
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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
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
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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
- A23K20/105—Aliphatic or alicyclic compounds
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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
- A23K20/158—Fatty acids; Fats; Products containing oils or fats
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K40/00—Shaping or working-up of animal feeding-stuffs
- A23K40/30—Shaping or working-up of animal feeding-stuffs by encapsulating; by coating
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
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Abstract
The present invention relates to the use of natural active substances selected from the group consisting of alpha-pinene, alpha-terpineol, cinnamaldehyde, dihydroeugenol, eugenol, meta-cresol and terpinolene in the manufacture of a feed composition for aquatic animals, especially for cold water fish as for example salmon, bream, bass and for warm water fish as for example carp, tilapia, catfish. More particular, this invention relates to the use of a substance as defined above for the improvement of the feed conversion ratio and/or daily weight gain in fish, for regulating the micro flora of the gut and for protecting the animal against infections caused by pathogenic microorganisms.
Description
The present invention relates to natural active matter is used to comprise fish and shrimp in manufacturing aquatic animal; Be particularly useful for the purposes in the feed of cold water fish (for example salmon, bream, perch) and warm water property fish (for example carp, Tilapia mossambica, catfish), said natural active matter is selected from the group of being made up of australene, α-terpineol, cinnamic acid, dihydro eugenol (dihydroeugenol), eugenol (eugenol), metacresol and terpinolene (terpinolene).
More specifically, the present invention relates at least two kinds of materials as defined above through regulating intestines microorganism species and/or exempt from feed conversion rate and/or the daily gain that the infection that is caused by pathogenic microorganisms is used to improve fish, the purposes that is used to reduce the death rate through watching for animals.
In addition; The present invention relates to novel fish meal composition; It comprises as two kinds of active component, preferably at least three kinds or four kinds of reactive compounds at least, and said reactive compound is selected from the group of being made up of australene, α-terpineol, cinnamic acid, dihydro eugenol, eugenol, metacresol and terpinolene.
Any compound, prepared product, mixture or composition that term " feed " or " fodder compound " expression are applicable to or are intended for use to be taken in by animal.
An important consideration in the culture fishery is a turnover rate.How soon turnover rate can be grown into by fish can be caught the size decision.As an example, atlantic salmon (Atlantic salmon) is raised and can be caught size needs 12 to 18 months from samlet (when atlantic salmon for the first time can be by the physiological stage when fresh water is transferred to seawater).Fast turnover has some good effects.At first, it helps cash flow.The second, it has improved risk management.Especially, high mortality is a material risk as far as the fisherman.
As everyone knows, unbalance microorganism species and/or can cause the death rate to increase by the infection that pathogenic microorganisms causes.The sashimi (raw fish) disease is common, and the possibility of illness outbreak is higher in long growing period.Because the accident when fishing net moves, or because bad weather causes that fishing net is broken, the risk that also has fish to run away.
For other domestic animals, it is known using antibiotic and vaccine to come the prevent disease development.In culture fishery, antibiotic use is so not extensive---and in cold water property culture fishery, this is because illness spread is very fast, and sick fish can not had plenty at least, and owing to the negative effect of discarded pastille feed to environment.It is widely used when vaccine can obtain, but they are not to develop to all diseases.
As the replacement of synthetic drug, in document, described and in animal feed, used plant extracts and essential oil.For example, patent documentation WO2004/091307 has described and in feed, has used polyphenol and other natural active matters, to increase the survival rate of the halogen worm (Artemia) after hatching.But WO2004/091307 is about the not record of selection of spendable compound.And in above-mentioned disclosed example, it is useful when hatching only that the application polyphenol reduces the death rate.
Therefore following demand is arranged in culture fishery:, come the prevent disease development thereby reduce the death rate through any preventative means (comprising antimicrobial acivity) on the enteron aisle level.
The present inventor finds that unexpectedly the material that defines like preceding text has the tremendous potential of in fish meal, using, and for example is used to improve feed conversion rate (FCR) and/or weightening finish and/or is used to regulate and control intestinal flora.And the inventor finds that unexpectedly these materials (hereinafter being also referred to as compound) also have the antimicrobial acivity that causes the death rate to reduce.The uniqueness of reactive compound of the present invention selects to allow first to control the many fish diseases that caused by many different pathogen.
Thereby; In first specific embodiment; The present invention relates in fish meal the method for using at least two kinds of reactive compounds that are selected from the group of being made up of australene, α-terpineol, cinnamic acid, dihydro eugenol, eugenol, metacresol and terpinolene to be used to improve feed conversion rate (FCR) and/or weightening finish and/or to be used to reduce the death rate, it is realized through regulation and control intestines microorganism species and/or through the disease of preventing to be caused by pathogenic microorganisms.For example; Show; The compound (for example: cinnamic acid, dihydro eugenol, eugenol and metacresol) that the present invention selects has been showed the good effect that suppresses Yersinia ruckeri growth; Said Yersinia ruckeri causes that sick (red mouth disease is found especially and caused high mortality for Enteric Redmouth, pathogenic microorganisms ERM) for red mouth in the salmon flying fish.
In the embodiment of replacement, australene and/or α-terpineol and/or cinnamic acid and/or dihydro eugenol and/or eugenol and/or metacresol and/or terpinolene are used to improve the animal feed digestibility and/or keep animal health through the support function of immune system.
In another embodiment; The present invention relates in fish meal the method for using at least three kinds of reactive compounds that are selected from the group of being made up of australene, α-terpineol, cinnamic acid, dihydro eugenol, eugenol, metacresol and terpinolene to be used to improve feed conversion rate (FCR) and/or weightening finish and/or to be used to reduce the death rate, it is realized through regulation and control intestines microorganism species and/or through the disease of preventing to be caused by pathogenic microorganisms.
In preferred embodiment, australene and cinnamic acid are used to improve the animal feed digestibility and/or keep animal health through the support function of immune system.
FCR can be based on the long test determination of following sashimi (raw fish), and the long test of said sashimi (raw fish) comprises: first processed group, wherein at least two kinds of compounds according to the present invention are added in the animal feed with the suitable concentration of every kg feed; With second processed group (contrast), do not add compound in animal feed.
As well-known, the FCR of improvement is lower than contrast FCR.In special embodiment, than contrast, FCR has been enhanced at least 1.0%, and preferably at least 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4% or at least 2.5%.
Be meant that like the term " intestines " that uses among this paper stomach and intestine or alimentary canal (being also referred to as digest tube) and its are meant the tract that dietary intake, digest food are extracted energy and nutrient and discharged residual waste in the multicellular animals.
Be meant like the term intestines " microorganism species " that use among this paper and inhabit in the intestines and natural microbial culture through helping suitably digestion to keep fit.
Term " regulation and control " ordinary representation relevant with the intestines microorganism species as using among this paper changes, operates, changes or adjust its function or state, i.e. non-therapeutic purposes in healthy and normal function animal.
Be meant through compound acquisition immunostimulation like the term " support function of immune system " that uses among this paper.
As the term " death rate " that uses among this paper be meant be included in the pond in the ratio compared of initial size of animal the existence animal in growth phase latter stage.It can challenge (challenge) test determination based on fish, and said fish challenge test comprises that it has the target of the death rate that in the group that is not processed, causes animal 40 to 80% by two groups of fishes of specific fish disease substance challenge.But; In feeding the challenge group that every Kg feed suitable concn is arranged according to the mixture of at least two kinds of compounds of the present invention; Than the group that is not processed, the death rate lowers at least 5%, and preferably at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or at least 50%.
Particularly, the present inventor finds that unexpectedly be effective according to compound of the present invention and composition thereof to many pathogenic microorganisms of cold water property and warm water property fish.Shown that australene, α-terpineol, cinnamic acid, dihydro eugenol, metacresol and terpinolene have the anguillarum to Vibrio---cause the inhibitory action of the shrimp pathogens of vibriosis.
Shown that australene, cinnamic acid, dihydro eugenol, eugenol, citrene and metacresol and terpinolene have the inhibitory action to Aeromonas salmonicida, Aeromonas salmonicida is the pathogen that causes the disease that is called dothienesis.
Shown that australene, α-terpineol, cinnamic acid, dihydro eugenol, eugenol, metacresol and terpinolene have the inhibitory action to the Edwardsellia tarda that causes systemic infection in the fish.
Shown that australene, α-terpineol, cinnamic acid, dihydro eugenol, eugenol and terpinolene have the inhibitory action of Dui Geshi galactococcus (Latococcus garvieae); The Ge Shi galactococcus is the cause of disease of lactic acid coccus sick (Latococcosis); In the several months in summer, be elevated to when surpassing 16 ℃ when water temperature, lactic acid coccus disease is the many fish species of influence and all causes a kind of acute disease of important economic loss in the two in seawater and fresh water culture fishery.
Cinnamic acid, dihydro eugenol, eugenol and metacresol are showed the good inhibitory action to the growth of Yersinia ruckeri; Yersinia ruckeri is the pathogenic microorganisms that causes red mouth sick (ERM), and red mouth disease is found in the salmon flying fish especially and caused high mortality.Shown that cinnamic acid and metacresol are to following inhibitory action:
(i) Vibrio salmonicida, it is a psychrophile, is the paathogenic factor of the cold water property vibriosis in the atlantic salmon.
(ii) Aeromonas hydrophila, it causes ulcer and bovine pasteurellosis.This pathogen to the simple antimicrobial of routine for example chlorine be that resistance is arranged very much.
(Iii) Photobacterium? Damselae, formerly known as Pasteurella? Piscicida, which is economically important marine fish, such as the Japanese amberjack, red grouper and American striped bass and kingfish aquaculture industry cause high losses pathogens.
(iv) Streptococcus iniae, it is high that cause a disease and be high fatal in the seawater fish: illness outbreak can be related with the 30-50% death rate.
Other water-borne pathogens are for example:
(i) Piscirickettsia salmonis is the paathogenic factor of piscirickettsiosis or salmons septicaemia (SRS),
(ii) Vibrio viscosus is renamed to Moritella viscosa recently, is the reason that is called as the disease of " ulcer in winter " on teiology,
(iii) Ich (parasite), one of most popular protozoon parasite of fish,
(iv) Vibrio harveyi causes the reason of photism vibriosis, and the photism vibriosis is the disease that influences the commercial aquaculture prawn,
The compound mixture that also can be described among the present invention suppresses.
In aspect second; The invention provides the fish meal composition, it comprises at least two kinds of reactive compounds that are selected from the group of being made up of australene (CAS 99-86-5), α-terpineol (CAS 98-55-5), cinnamic acid (CAS 14371-10-9/104-55-2), dihydro eugenol (CAS 2785-87-7), eugenol (CAS 97-53-0), metacresol (CAS 108-39-4) and terpinolene (CAS 554-61-0).Compound according to the present invention is commercial obtainable or can be easily be used known technology of the prior art and method preparation by the technical staff.
As the fish meal composition, compound of the present invention can natural obtainable extract or the form of extract mixtures or use separately or use with its mixture with the form of natural materials.
Comprise through solvent extraction (extract also is called as " extracting oil "), steam distillation (extract also is called as " essential oil ") or other compositions that obtains as the known method of technical staff like the term " extract " that uses among this paper.Suitable extraction solvent comprises alcohols, for example ethanol.
Term " natural " is understood that such material in this article, and it is by naturally occurring and from natural products, obtain or form through the compound of synthetic acquisition.
Reactive compound or natural materials or be further extracted into branch and can trace add.The example of specific examples of such components has: capsaicin, tannin, pipering, trimethylamine, 3,4-xylenol, furfuryl alcohol and composition thereof.
If as the mixture of at least two kinds of compounds of above-mentioned appointment is that preferably cinnamic acid and/or metacresol are used as the key component of mixture.Suitably, mixture contains metacresol and/or the cinnamic acid of 10-90% by weight, australene, the dihydro eugenol of 1-50% by weight of 1-50% by weight, and wherein said amount is based on the total of said component.The total amount of these active components can in wide boundary, change but finally with from 10 to 5000ppm, preferably 100 and 1000ppm between amount be used in the fish meal, said amount is based on that the dry weight of fish meal calculates.
Most preferred mixture like at least two kinds of compounds of above-mentioned appointment comprises cinnamic acid and australene as the key component of mixture.Suitably, mixture contains the australene of 30-70% by weight, cinnamic acid, metacresol, the dihydro eugenol of 1-20% by weight of 1-20% by weight of 30-70% by weight, and wherein said amount is with respect to the total of said component.The total amount of these active components can in wide boundary, change but finally with from 10 to 5000ppm, preferably 100 and 1000ppm between amount be used in the fish meal, amount is based on that the dry weight of fish meal calculates.
All compounds of the above-mentioned definition of this paper (reactive compound and extra composition) can use with the emulsifying surfactant combination.Emulsifying agent can advantageously be selected from quite hydrophilic those emulsifying agents, for example the polyglycerol ester of aliphatic acid, the for example propylene glycol ester of the castor oil acid of esterification or aliphatic acid, sugar esters or sugared glyceride type, polyethylene glycol, lecithin or the like.
Of the present invention preferred embodiment in; The mixture of reactive compound can contain 20% cinnamic acid by weight; 20% metacresol, 20% dihydro eugenol, 20% australene by weight by weight by weight; 3% trimethylamine by weight, 1.8% pipering and 4% furfuryl alcohol by weight by weight.Of the present invention another preferred embodiment in; The mixture of reactive compound contains the australene of 40 to 60 weight %, the cinnamic acid of 40 to 60 weight %; And can contain the trimethylamine of 1 to 5 weight %, the furfuryl alcohol of 1 to 5% pipering and 3 to 8 weight %.
The fish meal composition that will contain reactive compound is mixed in the fish meal into the carrying out that can describe like embodiment 1.The mixture of reactive compound directly is prepared as oil then, and said oil then mixes with the oil that is sprayed to as on the feed granules of in embodiment 1, describing.
The fish meal composition that will contain reactive compound is mixed in the fish meal or also can be realized through the premix of preparation active component and other suitable additives.This type of premix can comprise active mixture or natural materials or the extract of 2-10% by weight, other conventional additives of 0-40% (for example flavoring) and any conventional absorbability carrier of 50-98% by weight by weight.
Carrier can contain, for example the wood-fibred of 40-50%, tristearin, curcuma powder, powder from rosemary, lime stone, natural gum for example gum arabic, sugar and/or starch and the water of 5-15% by weight of 5-50% by weight of 1-3% by weight of 22-28% by weight of 4-5% by weight of 4-5% by weight of 8-10% by weight by weight.
Then for example vitamin C, mineral salt and other feed addictive compositions (yeast extract that for example contains nucleotides, glucan and other intestines microorganism species adjusting control agents) mix with vitamin with this premix; Finally can be contained 10-5000ppm with feed then, preferably the such amount according to active component of the present invention of 100-1000ppm or 100-500ppm is added in the feed.And composition of the present invention will preferably use with the yeast extract that contains nucleotides and glucan.
And, optional, the feed addictive composition is a colouring agent, for example carotenoid (for example beta carotene), astaxanthin and lutein; Aromatic; Stabilizing agent; Antimicrobial peptide; Polyunsaturated fatty acid; And/or be selected from: phytase (EC 3.1.3.8 or 3.1.3.26); Zytase (EC 3.2.1.8); Galactanase (EC 3.2.1.89); Alpha-galactosidase (EC 3.2.1.22); Protease (EC 3.4.); Phospholipase A1 (EC 3.1.1.32); Phospholipase A2 (EC 3.1.1.4); Lysophospholipase (EC 3.1.1.5); Phospholipase C (EC 3.1.4.3); Phospholipase D (EC 3.1.4.4) amylase, for example AMS (EC 3.2.1.1); And/or at least a enzyme in the 1,4 beta-glucanase (EC 3.2.1.4 or EC 3.2.1.6).
The example of polyunsaturated fatty acid is C18, C20 and C22 polyunsaturated fatty acid, for example arachidonic acid, DHA, EPA and gamma-linoleic acid.
The albumen and the moisture of 1-45 weight % and the proximate composition of lipid that have 20-60 weight % like the fish meal of describing among this paper.
In some concrete examples, fish meal contains:
---albumen, carbohydrate and lipid (for example fish meal, fish oil, blood meal, feather meal, poultry powder, chicken powder and or the powder of the other types that from other slaughterhouse discarded objects, produce),
---animal tallow (for example poultry oil),
---plant powder (for example soy meal, feather fan bean powder, peameal, bean powder, ground oil seeds and/or sunflower powder),
---vegetable oil (for example rapeseed oil, soybean oil),
---seitan (for example gluten or corn gluten) and
---the amino acid (lysine) of interpolation
In one or more.
Term " fish meal " as using among this paper comprises according to fish meal composition of the present invention and component as described above.Typically, fish meal comprises the fish meal as component.Suitably, fish meal is sheet or particle (for example extruding particle) form.
In the third aspect, the present invention relates to be used for the fodder compound of aquatic animal and relate to the purposes that said composition is used for feeding fish.This feed is particularly suitable for raising salmon, and it comprises atlantic salmon (salt lake salmon), other salmon species and trouts, and non-salmon for example cod, jewfish, porgy (sea bream) and eel.However, this diet may be kept for all types of fish, such as turbot, halibut, herring (yellow? Tail) and You fish.
Description and claimed invention are not limited in the scope of the disclosed specific embodiment of this paper among this paper, because these embodiments are intended to the explaination as some aspects of the present invention.The embodiment of any equivalence within the scope of the invention.Really, in this paper, show and describe those, multiple improvement of the present invention to those skilled in the art can be obvious from above-mentioned specification.This type of improvement also drops in the scope of appended claim.
Embodiment 1: the preparation of compacting fish meal
Main raw material(s) is ground and mixing.Then micro constitutent is added in the blender, and in preconditioner, homogeneous mixture is regulated through adding water and steam.This has begun the process of hotting plate in starch part (component of combination).Material is fed in the granulator.Material is compelled to through the grinding machine die orifice and is particle at the external fragmentation of die orifice.Water content be low and the feed drying not necessarily.Comprising that the extra oil according to fish meal composition of the present invention then is sprayed on the surface of particle, is quite fine and close as particle still, and total lipid content seldom surpasses 24%.The oil that adds can be fish oil or vegetable oil, for example the mixture of the mixture of rapeseed oil or soya-bean oil or vegetable oil or fish oil and vegetable oil.After oil coated, particle cooled off in cooler and packs.The fish meal of compacting contains 10 to 5000ppm the composition as describing in the present invention at last.
Embodiment 2: the method that is used to prepare extruding fish feed
Main raw material are ground and mixing.Add in the blender comprising micro constitutent according to fish meal composition of the present invention.In preconditioner, homogeneous mixture is regulated through adding water and steam.Also can add extra oil in this stage.This has begun the process of hotting plate in starch part (component of combination).Material is fed in the extruder.Extruder can be single screw rod or double-screw type.Because material rotatablely moves the further mixing of material quilt in extruder.Extra oil, water and steam can be added in the material in extruder.At the end of extruder, material has temperature that is higher than 100 ℃ and the pressure that is higher than environmental pressure.Material is compelled to the opening through the extruder die plate.Because temperature and pressure alleviates, some moisture evaporation (flash distillation) and material of extruding rapidly become porous.Through rotor line is cut into particle.Water content is quite high (18-28%) and particle thereby in drier, is dried to about 10% water content rapidly.
After drier, through oil is sprayed on the surface of feed, or through in oil, soaking feed, more oil can be added into feed.Air pressure is lower than and in the container of sealing of ambient pressure (vacuum plating) oil is added into feed and has superiority therein, and this makes the porous feed granules absorb more oil.Containing the feed that surpasses 40% lipid can this mode produce.After coating machine, feed is cooled and packs.As the method explained hereinbefore in, oil can add on some positions, and oil can be fish oil or vegetable oil, for example the mixture of the mixture of rapeseed oil or soya-bean oil or vegetable oil or fish oil and vegetable oil.
In order to grow and to keep the good health state, fish needs protein, fat, minerals and vitamins.The diet of carnivorous fish is a particular importance.At first, in the breed of carnivorous fish, full fish (whole fish) or crushing fish (ground fish) are used for satisfying the nutritional need of culturing fish.Be mixed with the crushing fish of the dried raw material (for example fish meal and starch) of multiple kind, be called as soft feed or semi-humid feed.Along with aquaculture becomes industrializedly, soft feed or semi-humid feed are pressed dry feed and substitute.It self is extruded dry feed gradually and substitutes.
Nowadays, extruding feed almost is general in the breed of many fish species (for example polytype salmon, cod, jewfish and porgy).
Important dietary protein origin in the dry feed of fish is the fish meal of different quality.Other animal proteins source also is used to the fish meal done.Thereby, use blood meal, bone meal, feather meal and the powder of the other types that from other slaughterhouse discarded objects, produce for example the chicken powder be known.These are more cheap than fish meal and fish oil usually.Yet, in some geographic areas, have in Feed Manufacturing, using these type of raw material to be used to produce the ban of the animal and the fish of food.
For example gluten, maize (corn) seitan, soybean protein, feather fan bean powder, peameal, bean powder, ground oil seeds, sunflower powder and ground rice also are known to use vegetable protein.
Embodiment 3: reactive compound according to the present invention is to Yersinia ruckeri; Vibrio anguillarum; Vibrio salmonicida; Aeromonas salmonicida; Aeromonas hydrophila; Edwardsellia tarda; Photobacterium damselae; Lactococcus garvieae; The assessment external test composition of the present invention of the effect of the survival of Streptococcus iniae is to Yersinia ruckeri; Vibrio anguillarum; Vibrio salmonicida; Aeromonas salmonicida; Aeromonas hydrophila; Edwardsellia tarda; Photobacterium damselae; The antimicrobial acivity of Lactococcus garvieae and Streptococcus iniae.
In test, used following organism, growth medium, condition of culture and appraisal procedure:
Bacterium: the pathogenic bacterial strains of all tests belongs to Centre for fish and wildlife health, Institute of Animal Pathology, bacterial strain collection place of University of Bern (Switzerland).
Suitable bacterium is dilution to be confirmed: from (Biomeheux, 24 hours old subcultures of the bacterium on Geneva) will be transferred among the aseptic NaCl, up to the McFarland value that obtains 0.5 in cultivating on a small quantity at the sheep blood agar.From this solution, in TSB, process 3,1.5 and 0.75% dilution factor having on the 96 hole flat boards of round bottom.100 μ l cumulative volumes are accepted in each hole.Under 22 ℃, after 24 hours, the evaluation bacterial growth.The dilution factor that causes covering the good spot in half the boundary line, round bottom hole is selected for experiment.
The mensuration of solvent effect: use ethanol (ETOH) to be dissolved in the test substances in the TSB agar.For measuring the possible effect of ethanol, in instrument connection, test the ethanol final concentration (0.1,0.05 and 0.025%) that calculates with the bacterium of variable concentrations, do not show any effect to bacterial growth.
The concentration of test substances: consider the meals concentration likely of 1000ppm at least and 2% day raise ratio, assessed external dosage range.Potential concentration in intestines is set up with maximum 0.1 μ l/100 μ l.Tested continuous 2 dilutions, this has caused the material final concentration (when transferring to average essential oil density) of 0.85 μ g/ml, 0.42 μ g/ml and 0.21 μ g/ml.
From every kind of material, the stoste that preparation is made up of 2 μ l materials, 18 μ l ETOH and 180 μ l PBS.
Dull and stereotyped preparation: respectively three parts of three concentration (0.21,0.42 and 0.85 μ g/ml) of test every kind of material.On each flat board, comprise blank (PBS) and the positive control of forming by the bacterium in TSB.And, comprise also that on each flat board other three of ETOH dilute each three parts.
Dull and stereotyped reading: 22 ℃ hatched in following 24 hours after, use 0 (do not have bacterial growth=on the bottom of flat board, do not have round dot) to 3 keeping the score of (normal growths=with the round dot round dot size relatively of positive control) read flat board.
The acquisition following result also is summarised in the table 1:
The suitable bacterial concentration that is used to test: 1.5% concentration of 0.5 McFarland value causes covering the clear visual spot in 2/3 round bottom hole.This concentration thus be considered to be suitable for the test.
ETOH is to the effect of bacterial growth: do not find the effect of the concentration of alcohol (1%, 0.5% and 0.05%) of any application.
The effect of test substances: the result is summarised in the table 1.Find out in three parts that big difference between three parts on the different flat boards, not find out big difference yet.All positive controls have shown clear visual bacterial growth in 48 hours.As from the pilot study desired, do not measure the negative effect of ethanol.Use cinnamic acid and australene to use metacresol and dihydro eugenol subsequently, found effect the wide spectrum of bacterium.
Table 1: 22 ℃ hatched in following 24 hours after, different materials is to the effect of the growth of Yersinia ruckeri, Vibrio anguillarum, Vibrio salmonicida, Aeromonas salmonicida, Aeromonas hydrophila, Edwardsellia tarda, Photobacterium damselae, Lactococcus garvieae and Streptococcus iniae.Value is represented each mean value of three parts separately 5 times.Nd: do not measure 3: not effect, 0: maximum growth suppresses.
Table 1:
Nd=does not survey
Embodiment 4: reactive compound according to the present invention is to the assessment of the effect of the survival of Vibrio anguillarum and Aeromonas salmonicida.
22 ℃ hatched in following 24 hours after, the combination of test different material is to the growth inhibiting effect of Aeromonas salmonicida and Vibrio anguillarum.Described in test condition such as the embodiment 3 and the test concentration be 0.21 μ g/ml, 0.42 μ g/ml and 0.85 μ g/ml.Table 2 and 3 value are represented 3 independent mean values of three parts on Aeromonas salmonicida and three of Vibrio anguillarum different flat boards respectively.3: expression is effect not, and 0: the expression maximum growth suppresses.
Separately or with the material of combined test is metacresol, cinnamic acid, australene, the dihydro eugenol.For two kinds of materials with 1: 1 ratio, for three kinds of materials with 1: 1: 1 ratio and for four kinds of materials with 1: 1: 1: 1 ratio prepares the mixture of material.
The result: all compounds and combination of compounds have been presented under the maximum concentration some inhibitory action to two kinds of pathogen, and observe to the obvious dose dependent of the effect of two kinds of bacterial species and shown in table 2 and the table 3.
The inhibition test result of table 2:Aeromonas salmonicida
The inhibition of table 3:Vibrio anguillarum
Embodiment 5: the composition of essential oil that comprises australene, cinnamic acid, dihydro eugenol and metacresol is to the effect with the death rate of the rainbow trout of Aeromonas salmonicida challenge
With rainbow trout foundation childhood, add to the effect of three dosage of composition of four kinds of essential oils of feed by the challenge of Aeromonas salmonicida experiment with test.
Fish: from the commercial fish culture field of Switzerland, obtain 600 young rainbow trouts.
Fish is raised condition: for domestication and the experimental feeding period before infecting, supported in the glass crawl that is equipped with running water current system and the 130L that continues ventilation from the fish that identical meals are handled, water temperature is maintained at 17 ± 1.0 ℃.
At infection time, be transferred in four glass crawls that are equipped with running water current system and the 38L that continue ventilation from the fish of each processed group.Water temperature is maintained at 17.5 ± 1.0 ℃.
Diet and raising: the basal diet that is used for rainbow trout childhood is with the production of 4mm diameter.From these particles, produced the chip (crumbles) of the size of suitable fish, and be coated on the chip with various dose through the essential oil composition that will be mixed with oil, produce a collection of feed of about 5-kg.Use peristaltic pump and mixed mixer to coat, to guarantee the even distribution of product.
3.3% ratio feeding fish every day with body weight.The diet ratio is adjusted in weightening finish to fish regularly.
Produce four kinds of experiments and used diet:
---diet A: the control diet that is not supplemented with essential oil composition
---diet B: the basal diet that is supplemented with the essential oil composition of 500ppm
---diet C: the basal diet that is supplemented with the essential oil composition of 2000ppm
---diet D: the basal diet that is supplemented with the essential oil composition of 4000ppm
Essential oil composition is following this: the australene of equivalent dosage, cinnamic acid, dihydro eugenol and metacresol.Before challenge with after challenging, feed all around up to the experiment end with diet with experiment.When experimental raising begins, before challenge and experiment when finishing to remaining fish record fish body weight.
Challenge experiment: in preliminary experiment,, measure challenge dose to identical fingerling crowd and under identical condition.Behind 4 time-of-weeks of experimental raising, 25 fishes are assigned in the glass crawl of 4 38L at random.Then with 1.2*10 by spectrophotometry
2The dosage of cfu/ fish will be from the fish anesthesia and the challenge of each experimental tank through lumbar injection Aeromonas salmonicida.Write down the day death rate of three time-of-weeks then.From the dead fish of article one of first glass crawl stand bacteriologic test with confirm to cause dead bacterial disease because of.Last in experiment, to the fish of external all survivals of sign evaluation of infecting and in addition from 6 fishes of each diet group, perform an autopsy on sb and bacteriological analysis.
The result of cumulative mortality lists in table 4:
Table 4: with respect to the mean value ± SD of the cumulative mortality of essential oil composition dosage
The result: it is normal and long to about 14g from 4g the experimental session fish that feed is taken in.
The result has disclosed at the dose-dependent effect with the essential oil in the rainbow trout of A.salmonicida. challenge.In accepting two groups of higher dosage, found minimum mean value, and in accepting the group of lowest dose level, found that comparison is according to higher slightly cumulative mortality.These results hint that the mixture of 4 kinds of essential oils in the diet has beneficial effect to the rainbow trout that infects with A.salmonicida..Difference aspect the bacterium existence in testing the fish of having found cumulative mortality and survival when finishing.
Embodiment 6: the essential oil composition that comprises australene and cinnamic acid is to the effect with the death rate of the rainbow trout of Aeromonas salmonicida challenge
Target: with rainbow trout foundation childhood, add to the effect of the australene and the cinnamic acid of feed with test with the challenge of Aeromonas salmonicida experiment.
Fish: obtain 600 young rainbow trouts from DSM Nutritional Products.
Fish is raised condition: for domestication and the experimental feeding period before infecting, supported in the glass crawl that is equipped with running water current system and the 130L that continues ventilation from the fish that identical meals are handled, water temperature is maintained at 14.5 ± 1.0 ℃.
Under the time of infecting, be transferred in four glass crawls that are equipped with running water current system and the 38L that continue ventilation from the fish of each processing.Water temperature is maintained at 13.5 ± 1.0 ℃.
Diet and raising: the basal diet that is used for rainbow trout childhood is with the production of 4mm diameter.From these particles, produced the chip of the size of suitable fish, and be coated on the chip with various dose through the essential oil composition that will be mixed with oil, produce a collection of feed of about 5-kg.Use peristaltic pump and mixed mixer to coat, to guarantee the even distribution of product.
3.3% ratio feeding fish every day with body weight.The diet ratio is adjusted in weightening finish to fish regularly.
Produce four kinds of experiments and used diet:
---diet A: the control diet that is not supplemented with essential oil composition
---diet B: the basal diet that is supplemented with the australene of 4000ppm
---diet C: the basal diet that is supplemented with equal dose australene+cinnamic acid of 4000ppm
Before challenge with after challenging, feed all around up to the experiment end with diet with experiment.
When experimental raising begins, before challenge and experiment when finishing to remaining fish record fish body weight.
Challenge experiment: in preliminary experiment,, measure challenge dose to identical fingerling crowd and under identical condition.Behind 4 time-of-weeks of experimental raising, 25 fishes are by in the glass crawl of random distribution to 4 38L.Then with 5*10 by spectrophotometry
3The dosage of cfu/ fish will be from the fish anesthesia and the challenge of each experimental tank through lumbar injection Aeromonas salmonicida.Write down death rate every day of three time-of-weeks then.From the dead fish of article one of first glass crawl stand bacteriologic test with confirm to cause dead bacterial disease because of.Last in experiment, to the fish of external all survivals of sign evaluation of infecting and in addition from 6 fishes of each diet group, perform an autopsy on sb and bacteriological analysis.
The result: it is normal and long to about 8g from 1 the experimental session fish that feed is taken in.
The result has disclosed that independent australene does not have effect but the composition of australene+cinnamic acid has positive effect: reduce by 15% the death rate than control treatment.The result of cumulative mortality lists in table 5:
Table 5: with respect to the mean value ± SD of the cumulative mortality of essential oil combination
Claims (15)
1. at least two kinds of natural active matters that are selected from the group of being made up of australene, α-terpineol, cinnamic acid, dihydro eugenol, eugenol, metacresol and terpinolene are used to improve the feed conversion rate of aquatic animal and/or the purposes of daily gain in fodder compound.
2. at least two kinds of natural active matters that are selected from the group of being made up of australene, α-terpineol, cinnamic acid, dihydro eugenol, eugenol, metacresol and terpinolene are used to regulate the purposes of microorganism species of the intestines of aquatic animal in fodder compound.
3. at least two kinds of natural active matters that are selected from the group of being made up of australene, α-terpineol, cinnamic acid, dihydro eugenol, eugenol, metacresol and terpinolene are used to reduce the purposes of the aquatic animal death rate in fodder compound.
4. according to each purposes of claim 1 to 3, wherein said aquatic animal is a cold water fish, for example salmon, trout, bream or perch.
5. according to each purposes of claim 1 or 3, wherein said aquatic animal is a warm water property fish, for example carp, Tilapia mossambica or catfish.
6. according to each purposes of claim 1 to 5, wherein said fodder compound contains australene, cinnamic acid, dihydro eugenol and metacresol.
7. according to the purposes of claim 6, wherein said fodder compound contains the concentration all australene between every Kg feed 10mg and 5g, cinnamic acid, dihydro eugenol and metacresol.
8. according to the purposes of claim 6, wherein said fodder compound contains the concentration all australene between every Kg feed 0.1g and 1g, cinnamic acid, dihydro eugenol and metacresol.
9. at least two kinds of natural active matters that are selected from the group of being made up of australene, α-terpineol, cinnamic acid, dihydro eugenol, eugenol, metacresol and terpinolene are used for treating and the purposes of the disease preventing to be caused by pathogenic microorganisms aquatic animal.
10. the fodder compound or premix compositions or the feed addictive that are used for aquatic animal, it comprises at least two kinds of natural active matters that are selected from the group of being made up of australene, α-terpineol, cinnamic acid, dihydro eugenol, eugenol, metacresol and terpinolene as main component.
11. according to the feed addictive of claim 10, wherein said feed addictive composition is to contain australene, cinnamic acid, dihydro eugenol or the metacresol natural extract as main component.
12. be used for improving the method at the feed conversion rate of aquatic animal, at least two kinds of natural active matters that wherein are selected from the group of being made up of australene, α-terpineol, cinnamic acid, dihydro eugenol, eugenol, metacresol and terpinolene are added in the animal feed.
13. be used for being reduced in the method for the death rate of aquatic animal; Wherein, at least two kinds of natural active matters that are selected from the group of being made up of australene, α-terpineol, cinnamic acid, dihydro eugenol, eugenol, metacresol and terpinolene are added in the animal feed.
14. according to the claim to a method of claim 12 or 13, wherein said fodder compound contains the concentration all australene between every Kg feed 10mg and 5g, cinnamic acid, dihydro eugenol or metacresol.
15. be used to improve feed conversion rate and/or the daily gain of aquatic animal and/or be used to regulate the microorganism species of intestines and/or be used to protect aquatic animal to exempt from the method for the infection that causes by pathogenic microorganisms; Said method comprises ingesting to feed; To animal at least two kinds of natural active matters that are selected from the group of being made up of australene, α-terpineol, cinnamic acid, dihydro eugenol, eugenol, metacresol and terpinolene of effective dose are provided, said active material exists as the feed ingredient of being ingested by animal.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| EP09165789 | 2009-07-17 | ||
| EP09165789.0 | 2009-07-17 | ||
| PCT/EP2010/060325 WO2011006993A1 (en) | 2009-07-17 | 2010-07-16 | Use of natural substances as feed additives for aquatic animals |
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| CN102469808A true CN102469808A (en) | 2012-05-23 |
| CN102469808B CN102469808B (en) | 2014-01-15 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201080032387.9A Active CN102469808B (en) | 2009-07-17 | 2010-07-16 | Use of natural substances as feed additives for aquatic animals |
Country Status (7)
| Country | Link |
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| EP (1) | EP2453758A1 (en) |
| JP (1) | JP5881175B2 (en) |
| CN (1) | CN102469808B (en) |
| AU (1) | AU2010272494B2 (en) |
| CA (1) | CA2767873C (en) |
| CL (1) | CL2012000096A1 (en) |
| WO (1) | WO2011006993A1 (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103815207A (en) * | 2014-03-14 | 2014-05-28 | 河南聚丰饲料科技有限公司 | Mixed feed for health-care Yellow River carp |
| CN105432972A (en) * | 2015-12-04 | 2016-03-30 | 全椒县城西畜牧养殖专业合作社 | Selenium-rich feed additive for weever |
| CN108289478A (en) * | 2015-10-13 | 2018-07-17 | 帝斯曼知识产权资产管理有限公司 | Include the feed addictive for aquatic animal of essential oil and lysozyme |
| CN108289469A (en) * | 2015-12-02 | 2018-07-17 | 日清奥利友集团株式会社 | Cladding fat or oil composition |
| CN109258984A (en) * | 2018-10-08 | 2019-01-25 | 天津农学院 | Application of the cinnamic acid with immune growth promoting function in Cynoglossus semilaevis feed |
| CN114009626A (en) * | 2021-10-21 | 2022-02-08 | 广东信豚生物科技有限公司 | Fish meal substitute containing yeast extract and aquatic animal premix composition |
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| NO20093460A1 (en) * | 2009-12-02 | 2011-06-03 | Ewos Innovation As | Feed composition for fish containing a semiochemical masking compound and use of the compound. |
| CN102217712A (en) * | 2011-05-27 | 2011-10-19 | 贺喜 | Pinene-containing feed or feed additive |
| CN103143553B (en) * | 2013-03-29 | 2015-05-20 | 广东大众农业科技股份有限公司 | Method for biological disposal of animal carcasses in river channel |
| KR101746273B1 (en) * | 2015-02-25 | 2017-06-12 | 양정자 | -carotene The feed additive for abalone including -carotene |
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| CN108424814A (en) * | 2018-03-20 | 2018-08-21 | 甘肃烟草工业有限责任公司 | A kind of preparation method of matrimony vine essence and the application in cigarette |
| EP3747280A1 (en) * | 2019-06-07 | 2020-12-09 | Lucta, S.A. | Antimicrobial combination for use in animal feeds |
| CA3197578A1 (en) * | 2020-12-04 | 2022-06-09 | Lucta, S.A. | A feed additive for increasing omega-3/omega-6 polyunsaturated fatty acids ratio in aquatic animals |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103815207A (en) * | 2014-03-14 | 2014-05-28 | 河南聚丰饲料科技有限公司 | Mixed feed for health-care Yellow River carp |
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| CN108289478A (en) * | 2015-10-13 | 2018-07-17 | 帝斯曼知识产权资产管理有限公司 | Include the feed addictive for aquatic animal of essential oil and lysozyme |
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| CN105432972A (en) * | 2015-12-04 | 2016-03-30 | 全椒县城西畜牧养殖专业合作社 | Selenium-rich feed additive for weever |
| CN109258984A (en) * | 2018-10-08 | 2019-01-25 | 天津农学院 | Application of the cinnamic acid with immune growth promoting function in Cynoglossus semilaevis feed |
| CN114009626A (en) * | 2021-10-21 | 2022-02-08 | 广东信豚生物科技有限公司 | Fish meal substitute containing yeast extract and aquatic animal premix composition |
Also Published As
| Publication number | Publication date |
|---|---|
| AU2010272494B2 (en) | 2015-04-09 |
| AU2010272494A1 (en) | 2012-02-09 |
| CN102469808B (en) | 2014-01-15 |
| CL2012000096A1 (en) | 2012-07-06 |
| JP5881175B2 (en) | 2016-03-09 |
| JP2012533289A (en) | 2012-12-27 |
| EP2453758A1 (en) | 2012-05-23 |
| CA2767873A1 (en) | 2011-01-20 |
| CA2767873C (en) | 2018-08-21 |
| WO2011006993A1 (en) | 2011-01-20 |
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