CN1454058A - Bioactive food complex, method for making bioactive food complex product and method for controlling disease - Google Patents
Bioactive food complex, method for making bioactive food complex product and method for controlling disease Download PDFInfo
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- CN1454058A CN1454058A CN01812813A CN01812813A CN1454058A CN 1454058 A CN1454058 A CN 1454058A CN 01812813 A CN01812813 A CN 01812813A CN 01812813 A CN01812813 A CN 01812813A CN 1454058 A CN1454058 A CN 1454058A
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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
Abstract
The invention relates to a bioactive food complex product, method for preparing a bioactive food complex product and method for controlling disease using probiotics and quorum sensing inhibitors such as inhibitory furanones and other bioactive compounds included in both the continuous and dispersed phases of a bioactive food complex product. The product is comprised of a solids-in-oil or an oil-in-solids emulsion forming a first emulsion that is itself emulsified in polymer forming oil-in-polymer or solids-in-polymer emulsion complex. The bioactive complex is formed of two emulsions with the first emulsion comprising the dispersed phase and a hydrocolloid polymer serving as the continuous phase. The second emulsion complex is then crosslinked to form a physically stable matrix. The bioactive food complex or the first emulsion of the bioactive food complex then serve to deliver different bioactive components including probiotic bacteria and quorum sensing inhibitor molecules to the digestive tract and environment of animals such as shrimp or fish or other livestock raised commercially to effectively control bacterial disease by a novel combination of mechanism including: competitive exclusion, direct inhibit, digestion of cell-to-cell signaling molecules and direct inhibition of homoserine lactone and (acyl) homoserine lactone regulated processes of pathogenic bacteria. Thus, effective disease prevention and control is accomplished through the novel combined delivery and use of probiotic bacteria and quorum sensing inhibitory furanones.
Description
Technical field of the present invention
The present invention relates to the bioactive food joint product, prepare the method for biological joint product and utilize probio and colony's quantity sensitivity (quorum sensing) inhibitor such as inhibition furanone (furanone) and other are contained in the method that bioactive compound in bioactive food joint product continuous phase and the decentralized photo is controlled disease.
The description of technical background
Shell-fish aquatic animal and have the aquaculture of fin fish the food of high value to be provided and in the international agriculture trade, to have belonged to field with fastest developing speed for the human consumption.The continuation of aquaculture development is subjected to the restriction of following factor: (1) lacks suitable commercial feed and (2) in the whole production phase in the incubation period of key and the phase of fostering, especially in the shrimp aquaculture process, and the destructiveness loss that brings by disease.
Hatching and the supply of fostering the common fresh and alive food organism of dependence of operation such as squid, polychaete worm, Artemia living beings, Artemia larva and microalgae come for cultivating and production aquaculture thing production aquaculture seedling.These foods carry a large amount of bacteriums usually, and these bacteriums may comprise pathogen such as vibrio bacterium, and may be the carriers that virus disease is propagated.The use of fresh and alive food organism increased dangerous and these pathogen that the incubation period catches may by seedling be transmitted to foster with culture device in.
In the shrimp aquaculture process mainly by bacterium, particularly the massive losses that causes of vibrio bacterium and the disease that causes as white spot virus (White Spot Virus) of virus causes heavy losses to many countries as this industries of countries such as China, Thailand, Indonesia, India, Philippine, Panama, Ecuador.The high density of incubation tank and hatching pond life has been aggravated the propagation of pathogen.Aquatic environment is suitable for bacterial growth owing to often adding the feed that is rich in albumen, and many bacteriums wherein may be caused a disease.These problems are because the interaction of microorganism and more and more serious, tentatively consider animal and environment thereof the intensive production scale from the clinicopathologia angle.When detecting malignant bacteria or virus, the culturist just adds Antimicrobe compound in feed and water; Even when pathogen was not obvious, many culturists also used antibiosis usually to prevent in large quantities.This causes increasing of vibrios, also may be increasing of other bacterium, and these bacterium have multiple antibiotic resistance and are the stronger pathogen of toxicity.Pathogen comprises vibrio harveyi, vibrio parahaemolytious, Vibrio splindidus, vibrio mimicus, comma bacillus, separates alginic acid vibrios, Vibrio anguillarum etc.
Utilizing beneficial microbe (probio) to replace malignant bacteria by competition process and direct inhibition is the remedial measure that is better than administration of antibiotics.Probio can be used to control the disease that is caused by vibrios and other Gram-negative bacteria and can be used to control and comprise other fishes and the shrimp disease that is caused by Aeromonas and Gram-positive pathogen, and described Gram-positive disease comprises streptococcus, meat bacillus etc.
These are used to culture fishery at present, but their efficient is different; Many products be not design at aquatic breed or be not the pathogen that is exclusively used in the purpose animal or will controls.In other words, they do not satisfy the definition of probio: the culture of the viable microbial of Cun Zaiing naturally helps animal health after this microbe passive thing is edible.
Usually, employed probio is added in the water, or in some cases, mixes rapidly with feed before using.Owing to having adopted high temperature to cause the death of viable bacteria in the preparation process of dried granulated meal, so probio can not add in the preparation process of multiple graininess and great majority extruding feeds.
WO9629392 has put down in writing by utilizing furanone (furanone) to suppress some Gram-negative bacteria, especially the method for the process of the endobacillary homoserine lactone adjusting of arc; Yet also nobody proposes to be suitable for the furanone employment mechanism that commercial size is cultured.
In the incubation period, can't effectively control micropopulation by food and water.Biology is sealed (bioencapsulation), promptly be used to increase and improved living food usefulness biological as seawater shrimps by the method for enriching described food organism nutrition with biological suspension, emulsion or other preparation that is fed with the compound of desired content to living food, Artemia belongs to biological nutrient inventory (Lavens etc., aquaculture and fishery management Aquaculture and FisheriesManagement) (1992)), next wherein said compound is used to feed the fishes and shrimps young.This technology has been used to bioactive compound such as antibiotic (Dixon etc., 1995, Journalof Aquatic Animal Health7:42-45) and bacterium (.1998 such as Gomez-Gil, Applied and Evironmental Microbiology64 (6): 2318-2322) pass to the aquaculture kind.Biology is sealed and is provided the effective ways that are used for the nutritional labeling in the food organism body and bioactive compound are passed to the carrier of purpose aquaculture kind.Yet biology is sealed to the breeding process of raising food organism and purpose kind in commercial hatching process and has been increased complexity, and the labour intensity of described breeding process is very high.
The commodity young is sold with living food thing substitute or supplements that dry feed has been used as the fish and the shrimp young, and wherein said feed comprises the particle and the tablet of albumen-crosslinked microcapsules, spray-dired aggregation, freeze-drying.Yet, very poor usually with these situations that do not contain the survival of the fishes and shrimps young that living food is food with biological young dry feed and/or growth.Therefore, the hatching of ocean fish and extra large shrimp can reduce but can not cancel the application living food and carry out seedling production with biology.
The relevant issues that prawn and fish are used young dry feed comprise: (a) physical instability in water, promptly join culture tank after feed granules decompose fast; (b) the high speed drop of nutritional labeling goes out, and promptly water miscible organic substance is lost in the groove water from complete feed granules/microcapsules.The physical decomposition and the drop of the young feed of selling go out to cause water quality very poor.Water quality inferiority is to the pressure root of the fishes and shrimps young and owing to the growth that has promoted pathogen has reduced health degree and the survival rate in the hatching.
The decomposition of young feed and drop go out degree can be reduced in the following manner: (a) strict seasoning more is insoluble in water thereby this method can increase the hardness of feed granules; (b) feed nutrients is wrapped up or is encapsulated in water-insoluble dressing or the matrix; Or (c) in young feed goods, adopt the feed ingredient of low aqueous solubility.Yet the cost that these method improvement water stabilities and reduction drop go out degree is to have lost the bioavailability of the not well-developed fishes and shrimps young of digestive system to nutritional labeling, and they can not obtain proper nutrition from these feeds.
Protein-crosslinking microcapsules (BP 79437454 and 2103568) are used biological as the living food that the dry powder commodity replace in the juvenile prawn food.Yet, the solubility nutrients that these capsules are a large amount of owing to drop has gone out to lose and can not replace algae alive and Artemia in the juvenile prawn breeding process fully.
WO87/01587 has put down in writing the formation of fat gel particle, thereby nutrition or pharmacy activity component are embedded in the liposome and this liposome is encapsulated in the hydrocolloid matrix.Described plasmalogen can effectively contain micromolecular, water miscible compound in the phosphatide microballoon and described hydrocolloid matrix can be protected described plasmalogen.Yet, the fat gel particle can not contain enough nutritional labelings because plasmalogen is loaded the little and more crisp immobilized artificial membrane of volume and constitute young feed completely, when mixing with required other feed nutrient of complete young feed, described immobilized artificial membrane may break.
(marine biology (Marine Biology), 115 (4): 635-642 (1993)) described composite micro-capsule (CXM), these microcapsules can significantly reduce low-molecular-weight water soluble compound drop from young feed and go out for Villamar and Langdon.Be embedded in the bigger gel pearl by the fat wall microcapsules (LWM) that will contain aqueous solution content material and prepare CXM, the big pearl of described gel contains feed nutrient and forms by mosanom with the gelatin that calcium carries out ionomer.LWM can be retained in little water soluble molecules among the CXM, but its load volume is little and the preparation difficulty.
Young feed CXM is with solving the physics water stability and because drop goes out to cause the problem of nutritive loss, but can not transmit bioactive compound or control disease.Biology is sealed and is used for improving living food with biological nutritive value and bioactive compound is delivered in the purpose kind, but this technology has increased complexity and cost for again the very complicated fish and the animal husbandry of crustacean larvae.
U.S. Pat 5698246 has been improved the (marine biology (Marine Biology) by Villamar and Langdon development, 115 (4): CXM notion 635-642 (1993)), by by adopting lipid such as fish oil parcel or the dried nutrient fodder of dressing will describedly be wrapped then or being formed LWM and eliminate complexity simplification technical process with sealing as the bacterium of the Bacillus of interior-probio by the dried nutrient fodder of dressing.The load mixture of the feed of dressing and bacterium is encapsulated in a kind of hydrocolloid matrix as carrying out in the mosanom of ionomer with calcium.Stablize entrapped compound by being suspended in a kind of liquid medium that helps to preserve then, described medium contains the probiotic bacteria as " outer probio ".In this liquid, Gl road and outer probio that the interior probio of young feed is designed to together to enter with nutritional labeling purpose shrimp or fish are designed to keep and grow in the water of grow shrimp or fish.
Though, United States Patent (USP) 5,698,246 by invention new liquid feed the conventional feed goods to aquaculture have carried out tangible improvement, but the method that this patent did not both provide the control disease is the technology of development preparation CXM also, but has adopted the method for simple parcel or the contained material of dressing to substitute LWM and the method for transmitting probiotic bacteria is provided and describes and be used for the method that disease is controlled in aquaculture.
U.S. Pat 5,776,490 relate to and are contained in compound, the crosslinking protein microcapsules rather than the LWM in the hydrocolloid matrix of the calcium alginate of ionomer and gelatin improves CXM by utilization.This invention has been used to nutrients and medicine passed to the fin fish young but has not been provided for the method for disease control in the aquaculture process.
I phase USDA/SBIR authentication code 93-33610-8500 (1993) has put down in writing the method that is used to prepare water stability aquatic animal feed, thus this feed be lower than process under 85 ℃ the low temperature optimized the bioavailability of aquatic animal to nutritional labeling.By in water, mixing slurries and the mosanom that contains feed ingredient, then described mixture cold extrusion or injection mould are prepared gel strips, this gel strips and calcium chloride carry out crosslinked and carry out drying at low temperatures, have so just made described water stabilize feed.In addition, thus this technical description the dry described slurries of employing vacuum drums make and the calcium cross-linked thin slice of chlorination.Thereby this technology utilizes low temperature process to avoid heating to nutraceutical destruction, and this nutrients prepares for aquatic animal, but this technology is not mentioned the transmission of bioactive compound or the control of disease.
WO95/28830 has put down in writing, and a kind of " environment temperature method " is 3-156 ℃, and this method comprises that mixed algae hydrochlorate, feed ingredient and water prepare slurries, thereby these slurries contact formation water stability aquatic animal feed granules with bivalent cation.WO95/28830 does not relate to composition and method is carried out any improved method.
U.S. Pat 6,024,983 have put down in writing the microcapsule compositions that is used to transmit bioactive compound in the medical application that the reinforcement animal immune is replied.This invention relates to a kind of mixture, and the biocompatibility microcapsules of the one-level matters of containing biological activities of this mixture are encapsulated in the biocompatibility microcapsules of secondary matters of containing biological activities.This invention provides immune response to the animal or human's class disease antigen from flu antigen to gonorrhea-antigen.The purpose of using this invention is to use this bioactive compound to having the ripe immunity and the infected animal of constant temperature system.Yet this invention is not used to shrimp, has the animal of nospecific immunity system, and since relevant with biomedical sector, be not provided for the central method of controlling disease of aquaculture.
In another industry, utilize the microencapsulation of hydrocolloid microcapsules to be used to fixedly comprise the biological active agents of hybridoma (US5,116,747, US4,942,129) and pancreas islet (US4,806,355).Described cell fixation is used for biomedical purpose and is included in a large amount of biologic products of cultivating manufacturer's industry consumption of bioreactor in the gel pearl, perhaps is used for carrying out disease treatment in the animal body by medical treatment implantation people or non-aquaculture.
Also have in the other industry application, in order to prepare the cellular biomass that is used for the dairy products fermentation, utilize the calcium alginate microencapsulation with the BA lactococcus lactis subsp be fixed on the gel pearl (Morin etc., Applied and Environmental Microbiology, 58 (2): 545-550, (1992)).
In research is used, Chevalier and de la Noue (Enzyme Microb.Technol., 9; 53-56, (1987)) improved the production of enzyme of AMS by the carragheen gel pearl that in airlift fermentor, the biologically active hay bacillus is fixed in ionomer.
In animal breeding, the bacterium of Lactobacillus, Bacillus and other genus is to be used as directly edible microorganism usually and to add probio in the feed to, the growth that this probio can be used for strengthening the nutraceutical ability of digestion and can suppress potential pathogenic microorganisms in the livestock enteron aisle by enzyme is especially in pressure state following time.The improvement of animal health situation is normally owing to secrete antibacterial material that the inhibition pathogen grows and activate specificity and nonspecific immune response by probio in enteron aisle.
In aquaculture, to aquatile, especially young form is transmitted as the bioactivator of probio etc. can not be by conventional feed and realize the decomposition rapidly in water of these active materials smoothly, have very high drop and go out speed, or in preparation process, experienced coarse processing.Although U.S. Pat 5,698,246 and US5,776,490 represent the aquatile feed, the especially improvement of fishes and shrimps young technical field of feed, but these still belong to Villamar and Langdon (marine biology (Marine Biology), 115 (4): the conversion step of the method that is worked out 635-642 (1993)), the disease control method in the aquaculture process is not described.
The result of study of ocean fish is shown that nature is present in several bacteriums in the enteron aisle and produces in the casing slime that the material that suppresses bacterial pathogens such as Vibrio anguillarum etc. and these bacteria beneficial can adhere to fish and the (Olsson etc. that grow therein, Applied and EnvironmentalMicrobiology, 58 (2): 551556, (1992)).
The invention that this paper put down in writing is provided at Villamar and Langdon (MarineBiology), and 115 (4): carry out improved composition of matter on 635-642 (1993) basis and the not method that is used to control bacterial disease of record of other documents is provided.
General introduction of the present invention
The invention provides the composition that is used for hydrobiological bioactive food compound, prepare the method for bioactive food compound and raise hydrobiological method with described bioactive food compound, described aquatile includes but not limited to crustacean, mollusk and the fin fish young, postlarva, childhood and adult form is arranged that the present invention also is provided at the method for control bacterial disease in the aquaculture process.
The invention provides and in preparation process be added in feed with the gemma form selected probio and the mechanism of throwing something and feeding and making these probios activate and sprout when animal maybe will be added in pond or the groove water when described probio is provided simultaneously.And this method combines the method for controlling pathogen by the expression of gene that suppresses adjusting Disease-causing gene activity.The combination of these two kinds of methods is more effective and have a commercial viability than every kind of single method.Select to produce the probiotic bacteria of enzyme, the antibiotic that this enzyme had not only been degraded acyl homoserine lactones that homoserine lactone but also degrade produces by vibrios and selected probio to produce to suppress the vibrios growth or kill vibrios.
Thereby the present invention's record passes to commercial domesticated animal with the inhibition furanone to be suppressed in the Gram-negative thalline, especially the practical approach of the endobacillary homoserine lactone regulation process of the arc of Zhi Binging.The present invention with those inhibition compounds in animal feed and animal stable breeding system transmission and the transmission of probio and other bioactive compound combine.
Therefore, the invention provides in aquatic animal control and/or prophylactic method, comprising that the lively thing of feedwater is thrown something and fed contains the composition of at least a probiotic bacteria and at least a inhibition or modulating compound.In this method, the pathogenic of bacterium can be inhibited by the combination of following coefficient three kinds of mechanism:
A. utilize probiotic bacteria such as bacillus to control pathogen by the following method: by competitive exclusive method as competition food and space, thereby and usually reduce the expression that its number suppresses colony's quantity impression and suppresses Disease-causing gene by direct inhibition as the antibiosis that produces sterilization or antibacterial substance such as anti-Gram-negative and Gram-positive pathogen by original position;
B. utilize probiotic bacteria such as bacillus to experience molecule and suppress the expression of Disease-causing gene in Gram-negative bacteria and Gram-positive pathogen by colony's quantity of secreting by the enzyme degraded pathogen of probio secretion.；
C. use inhibition or modulating compound such as furanone so that the expression of the Disease-causing gene that in Gram-negative pathogen such as vibrios, suppresses to regulate by homoserine lactone and acyl homoserine lactones.
In an embodiment of described bioactive food compound, the emulsion that contains first kind of oil bag solid that lipophilic bioactive compound forms as the inhibition furanone that is dissolved in the lipid forms continuous phase with the dry feed composition of the decentralized photo of stabilized emulsion as described in forming and other bioactive compound such as selected probiotic bacteria.Described stabilized emulsion, emulsion-1 provide in decentralized photo and continuous phase that aquatic animal normally survives, required bioactivator and essential nutritional labeling in the g and D.Emulsion-1 itself is decentralized photo by hydrocolloid polymer emulsification, thereby hydrocolloid polymer is carried out ion gel formation bioactive food compound then.In following disclosure, hydrocolloid is the biodegradable polymer that the continuous phase of other form of the polymer of preferred type and second kind of emulsion can comprise crosslinking protein or other form.Described bioactive food compound contains the bacterial spore that essential nutrients and/or bioactive compound can be provided, the bacteriotrophy cell, bacteria cell wall, yeast cells, yeast extract, yeast cell wall, alga cells, medicament, enzyme, invertebral living creature embryo or the biological invertebral living creature of sealing, described must nutrients and/or bioactive compound help promoting the absorption of nutritional labeling and improve the efficient of assimilation, strengthen immune response and suppress the alimentary canal of aquatic animal and pond or tank or other are raised the growth of the intrasystem pathogenic microorganisms of hydrobiological stable breeding.
Described bioactive food compound can not need drying and preserve with the aneroid semisolid of being made up of microcapsules or pearl, wet pastel or with forms such as wet bar, particle, thin slices, perhaps carries out freezing preservation by employing antifreezing agent (cryopreservative).Described bioactive food compound can directly be added in the aquiculture animal stable breeding system so that eaten by aquatic animal.In one embodiment of the invention, described bioactive food compound or first kind of bioactive food compound can be used as in the face coat or aquatile feed that dressing is added to graininess or extruding of aquatile feed of graininess or extruding.
Novel two-emulsion method makes described bioactive food compound.In one embodiment, emulsion-the 1st is by the bioactivator that forms decentralized photo and powdery nutrient fodder with form the oil bag solid emulsion that the edible oil of the lipophilic bioactive compound that contains dissolving of continuous phase is formed.Emulsion-the 2nd, a kind of emulsion of polymer bag oil, wherein emulsion-1 as decentralized photo and hydrocolloid polymer as continuous phase.Thereby emulsion-1 contains the feed nutrient that is dispersed in the oil and bioactivator and it is formed emulsion-2 by the emulsification of hydrocolloid polymer institute.In one embodiment of the invention, the bioactivator that contains fat-soluble inhibition furanone is mixed in the continuous phase of emulsion-1.In another aspect of this invention, bioactivator that may break or damage in the process that forms emulsion-1 such as invertebral living creature or embryo or probiotic bacteria vegetative cell can be directly embedded in the emulsion-2 by slight mixing.Emulsion-2 is become to combine biological active material and the nutritional labeling and the matrix that is embedded in the physically stable of the complete bioactivator in the emulsion-2 of emulsion-1 by ionomer or complexing then.In ion gel method, described bioactive food compound can form microcapsules or pearl, bar, particle, tablet or other geometry.
The edible oil that is used as the continuous phase of emulsion-1 can be the mixture of fish oil, refined fish oil's product or fish oil, fish oil product and vegetable oil.
Typical oil comprises herring oil, salmon oil, long tail anchovy oil, pilchard oil, tunny fish oil, mackerel oil, thin squama cucumber fish oil, Squid Oil, pollack fish oil, cod-liver oil, edible fish oil additive such as Promega (Warner-Lambert company), soybean oil, safflower oil, corn oil, palm-kernel oil and other edible oil.In addition, described oil or oil mixture have added inhibition furanone, lecithin, cholesterol, emulsifying agent such as Santone (Van Den BerghFood Ingr.), antioxidant such as ethoxyquin, fat-soluble A, D, E and K, β carotene and astaxanthin pigment.
After inhibition furanone and other lipophilic bioactive compound, lecithin, cholesterol, emulsifying agent, antioxidant, liposoluble vitamin and astaxanthin are mixed in the oil, form emulsion-1 thereby the dry powder feeding composition is emulsified in oil mixture, this emulsion is the emulsion of oil bag solid.Of the present invention preferred aspect, solid to the proportion of oil from about 0.01: 1 to 100: 1, preferably 0.1: 1 to 3: 1.Therefore, depend on the relative concentration of solid and oil, the composition of emulsion-1 can wrap solid from oil and change to solid bag oil.
Two types emulsion is in protection scope of the present invention.In following disclosure, when milkiness also is recited as oil bag solid emulsion, should be understood to described description and also be applicable to solid bag oil emulsion and except as otherwise noted, should be understood to bioactive compound and both be comprised in two types of decentralized photos in the emulsion and also be contained in its continuous phase.
In a preferred embodiment of the invention, the bioactivator that comprises bacteria cell wall, bacterial cell or gemma, yeast, yeast cell wall, yeast extract, alga cells, medicament or enzyme is contained in the solid phase of emulsion-1.The oil phase of emulsion-1 is as the medium that comprises lipophilic bioactive compound such as inhibition furanone, fat-soluble hormone, fat-soluble chemical attractant or chemical stimulus or other lipophilic bioactive compound and as the hydrophobicity internal layer coating of mash feed composition and bioactivator.Emulsion-1 provides essential fatty acid, fat-soluble nutriment and vitamin and pigment also as the nutrient source of aquatic animal.
With liposome, LWM or known be wrapped/different by the material of dressing, emulsion of the present invention-the 1st all contains the oil bag solid of important biomolecule activity or the novel emulsion of solid bag oil in continuous phase and decentralized photo, its preparation is simple, stable, and can provide and reduce barrier that water soluble molecules goes out to the water environment drop from the powdery nutrients with from bioactivator and the restriction that can not be subjected to the such little load volume of liposome or LWM.Emulsion-1 physics and chemical aspect be different from U.S. Pat 5,698, the parcel of record or the straightforward procedure of dressing in 246, and representative is to Villamar and Langdon (Marine Biology, 115 (4): the improvement of described complexing microcapsules (CXM) 635-642 (1993)).
Thereby contain the bioactivator of fat-soluble and solid and the emulsion-1 of feed ingredient and itself be emulsified into hydrocolloid polymer dispersion formation emulsion-2, the emulsion of polymer bag oil or polymer bag solid.That is, the decentralized photo of emulsion-2 is made up of emulsion-1 and the continuous phase of emulsion-2 is made up of hydrocolloid polymer.Emulsification may can be embedded in the emulsion-2 by slight mixing by damaged invertebral living creature embryo or invertebral living creature, bacteriotrophy cell or other material.Then emulsion-2 be used for the ion of ionomer hydrocolloid polymer and contact.Ionic gelization is used for forming the stable matrix of the continuous phase of emulsion-2, and it comprises the decentralized photo of emulsion-2 and by the material of embedding, forms the bioactive food compound.
In one embodiment, described bioactive food compound forms semisolid wet paste and preserves by moisture (Aw) is dropped to 0.8 having in the presence of antibiotic medium such as calcium chloride or sodium chloride brine and mould inhibitor such as glycerine, propane diols, propionic acid (proprionic acid) or other anticorrisive agent.In addition, described bioactive food compound can be by adopting antifreezing agent such as methyl-sulfoxide (DMSO), glycerine, butylated hydroxytoluene (BHT), sucrose, gossypose, sweet mellow wine (manitol), ethylene glycol, propane diols, methyl alcohol, 1,2-propane diols, 1,3-butanediol, 2, the 3-butylene glycol carries out freezing preservation.
Detailed description of the present invention
Gemma and dried vegetative cell with inhibition furanone, probio such as bacillus in preparation process are incorporated in the bioactive food compound, wherein select for use to produce special inhibition gram-negative pathogens such as the antibacterial material of vibrios and the probio of enzyme.Then the bioactive food compound directly is added in the food of aquatile kind of aquaculture or in the animal stable breeding system.The emulsion-1 of described bioactive food compound and bioactive food compound is added respectively or jointly as the active component of the outer dressing of traditional animal feed, and described traditional animal feed comprises aquatile feed granular or extruding.
Described bacillus species is that the basis is selected with the antibiotic that produce to suppress or kill malignant bacteria such as vibrios.Of the same race or different bacillus species similar they can produce and suppress the enzyme that the intercellular signal in pathogen such as the vibrios transmits by degraded or digestion signaling molecule such as homoserine lactone (HSL) and (acyl group) homoserine lactone (AHL) and be the basis selection.
In one embodiment of the invention, the signaling molecule of described degraded or digestion pathogen and the inhibitory action that takes place is united in shrimp, fish, mollusk and other aquatic animal enteron aisle and competition nutritional labeling and binding site on the shell of their outer surface such as crustacean, on the molluscan soft tissue and rete malpighii fish, thus not only suppress the expression of Disease-causing gene but also reduce the quantity of these lip-deep pathogenic bacteria such as vibrios.
In the preparation process of described bioactive food compound and conventional animal feed, heat simultaneously and pressurization is enough to start the sprouting of (activations) bacillus gemma, need only preparation temperature and preferably maintain below 100 ℃.Be provided at alanine and other amino acid that can activate sprouting after bioactive food compound or the feed generation aquation in the particle by the solubility content of microorganism such as yeast extract are added to, further promote its growth.
In a preferred embodiment of the invention, in the emulsion-1 or the bioactive food compound in the gemma of bacillus species and the inhibitor of dried vegetative cell and other bioactive compound, especially signaling molecule such as HSL and the AHL special group quantity perception of regulating be used as outer dressing and add on the feed of aquatile or extruding.By competition and the inhibition of Bacillus probio, thereby the quantity that has reduced Gram-negative in enteron aisle, body surface and/or the environment of animal and Gram-positive pathogen has suppressed the population quantity perception of pathogen and has suppressed the activation of the Disease-causing gene of these pathogen owing to the minimizing of pathogen quantity.The present invention prevents that the population density of pathogen from surpassing the lowest threshold that activates Disease-causing gene.By utilize competition nutrition of the present invention, space and directly the probio of pathogen kill the quantity of pathogen is remained on the low-level regulatory gene that suppresses group sensibility, do not produce disease thereby Disease-causing gene is not expressed.
Owing to the invention provides the most effective combined method or the mechanism of playing a role jointly, so method of the present invention is different from other method.If for example using inhibition furanone or HSL and AHL to regulate molecule separately is damaged or is digested and do not get rid of or directly suppress pathogen by probio of the present invention is competitive, the growth of pathogen just can not get suppressing so, and just suppressed the expression of Disease-causing gene, therefore, if stop the supply of inhibition furanone suddenly, if for example the culturist has used up the feed of handling through furanone, group sensibility then occurs and will secrete virulence factor, the result causes disease and causes animal dead.Bacillus has the effect of two aspects: the enzyme that (1) is produced by bacillus will be degraded by the expression of the HSL of pathogen such as vibrios generation and AHL and inhibition Disease-causing gene; (2) quantity of pathogen such as vibrios is owing to the competition of inhibitory action and food and other resource is reduced, therefore, there is not the population of specific quantity in low population density meaning, and promptly the quantity of described pathogen is too low so that HSL or AHL compound concentrations are too low and invalid.The result is that Disease-causing gene can not be expressed.Can not cause disease.
Therefore probio competition nutrition suppress the quick growth of pathogen.When pathogen contains the antibiotic that produced by probio by the gene of resistance, use described probio not imitate separately.Of the present invention one than other process embodiment preferred in, when probio and specific population quantity perception mechanism were used together, the probiotic bacteria of low quantity just can be effective, thus make product preparation simply and cost is low.And the antibiotic that the more impossible Bacillus bacterium to all uses of a kind of pathogen produces produces resistance, but also the effect of the enzyme of the pathogen signaling molecule of avoiding degrading.
In preferred embodiments, emulsion of the present invention-the 1st, the emulsion of oil bag solid, this emulsion is made up of the nutrient fodder of bioactivator that forms decentralized photo and powdery and the edible oil that is dissolved with lipophilic bioactive compound that forms continuous phase.In a preferred embodiment of the invention, described oil phase contains inhibition furanone and other biologically active fat-soluble compound.Described oil formulation prepares by following one or more are mixed in the edible oil: weight percentage is about lipophilic bioactive compound such as the inhibition furanone of 0.0001-50%, weight percentage is about emulsifying agent such as the Santone (Van DenBergh Food Ingr.) of 0.5-5%, weight percentage is about the lecithin of 1-10%, weight percentage is about the cholesterol of 1-10%, weight percentage is about antioxidant such as the ethoxyquin of 0.01-0.05%, about 2000 to 8000IU the vitamin A of every kg of oil, about 1000 to 4000IU the vitamin D of every kg of oil
3, about 2000 to 8000IU the vitamin E of every kg of oil (alpha-tocopherol), about 20 to 80mg the vitamin K of every kg of oil (menadione), about 2000 to 8000IU the beta carotene of every kg of oil and about 500 to 2000mg the astaxanthin of every kg of oil.
Typical edible oils comprises refining herring oil, salmon oil, long tail anchovy oil, pilchard oil, tunny fish oil, mackerel oil, thin squama cucumber fish oil, Squid Oil, pollack fish oil, cod-liver oil, edible fish oil additive such as Promega (Warner-Lambert company), soybean oil, safflower oil, corn oil, palm-kernel oil and other edible oil.Described oil product contain total concentration greater than the omega-fatty acid of about 1% weight percentage, total concentration greater than the eicosapentaenoic acid (EPA) of about 0.1% weight percentage and total concentration DHA (DHA) greater than about 0.1% weight percentage.In order to prepare emulsion-1, before adding powdery nutrient fodder and bioactivator, described oil mixture is heated to about 25-40 ℃.
On demand, for different stages life cycle or the different aquaculture application that different aquatile kinds, certain species are provided strengthens when cultivating pool transports defending and required essential COF, energy and the bioactivator of immune stimulation from hatching as the back young or fry as conventional raising or in unfavorable period, can adjust the solid compositions of emulsion-1, described solid compositions is made up of powdery nutrient fodder and bioactivator.Can realize using more in a wide range the bioactive food compound of animal to the solid compositions adjustment of following emulsion-1.
The solid phase of emulsion-1 is made up of in the following ingredients one or more: weight percentage be about 0.1-95% bacterial spore, bacteria cell wall, bacterial cell, include but not limited to Bacillus acidi lactici, bacillus, streptococcus, Bifidobacterium, leukonid (Leuconostor) and monad (Alteromonad) alternately; Weight percentage be about 0.1-50% yeast extract, yeast cell wall, food yeast, brewer's yeast, include but not limited to the yeast cells of torula and Phaffia yeast; Alga cells goods, alga cells that weight percentage is about 1-80% include but not limited to flagellate, Schizochytrium, Du Shi algae, Chaetoceros, Tetraselmis, Skeletonema Greville, Nannochloropsis, hailian seaweed, brown algae, Isochrysis galbana, Paylova; What weight percentage was about 0.01-50% includes but not limited to antibiotic such as Sarafin, Repirinast, the medicament of terramycin; Mash feed, its concentration are conditioned and are used for satisfying the animal diet followed needs, comprise that weight percentage is about the animal protein product of 0-95%; Weight percentage is about the plant protein products of 0-95%; Weight percentage is about the birds, beasts and eggs product of 0-25%; The cobalamin of about 40-60mg/kg; The D-biotin of about 5-20mg/kg; The D-pantothenic acid of about 250-350mg/kg; The folic acid of about 10-30mg/kg; The L-ascorbic acid of about 1000-4000mg/kg-2-polyphosphoric acid fat (STAY-C, ascorbic stable form); The inositol of about 3000-4000mg/kg; The nicotinic acid of about 600-800mg/kg; The p-aminobenzoic acid of about 350-450mg/kg; The pyridoxine hydrochloride of about 40-60mg/kg; The riboflavin of about 125-175mg/kg; The thiamine chloride of about 50-80mg/kg; The Choline Chloride of about 6500-7500mg/kg.
By with solid constituent with about 0.01: 1 to 100: 1, preferred 0.1: 1 to 3: 1 ratio is mixed into and prepares emulsion-1 in the oil mixture, and this emulsion-1 depends on that the relative concentration of described solid and oil ingredient both can be that oil bag solid emulsion also can be solid bag oil as turbid liquid.Emulsion-the 1st, stable and separation that phase does not take place 20-25 ℃ of following the preservation at least 3 hours.
According to the applicable cases of described bioactive food compound, with about weight percentage of 5 to 65% emulsion-1 is mixed into and prepares emulsion-2 in the hydrocolloid polymer.For juvenile prawn, the weight percentage that preferred composition is the emulsion-1 in the hydrocolloid polymer is about 35-45%.The continuous phase of emulsion-2 is about the 0.5-4.0% mosanom by the weight percentage that forms in 45-85 ℃ deionized water or κ carragheen polymer is formed.Thereby described alginates or carragheen polymer can mix to have increased and be used for configuration and the bond strength that aquatile carries out the site of proteolytic digestion and improved gel-type vehicle with dissolving in the weight percentage of about 0.1-3.5% and the water or the gelatin, zein, polylysin, poly arginine, shitosan, Arabic gum or the tracasol goods that disperse.Relative polypeptide of described alginates or carragheen or protein such as gelatin or other hydrocolloid or natural gum are about 2: 1 to about 10: 1 as the ratio of tracasol or shitosan.
When mosanom is used as the continuous phase of emulsion-2, thus water-bearing media is adjusted to about pH12 in case guarantee the alginates molecule electronegative with the calcium formation calcium alginate matrix that reacts.For κ carragheen polymer, potassium is preferred ion, thereby forms potassium carragheen compound with this polymer reaction.When with calcium or potassium complexing, contain emulsion-2 continuous phase polymer or polymeric blends is used for sealing or embedding emulsion-1.
Another aspect of the present invention, before forming the ionic gel-type vehicle, by will frangible material with the weight percentage of 1-50% being mixed into gently in the emulsion-2 and biological active component that will be more frangible such as invertebral living creature embryo or invertebral living creature are embedded in the emulsion-2.Described frangible material can comprise metazoa, protozoan or other microorganism or the plant or the animal tissue of any suitable work, freezing or freeze-drying, keeps complete relatively intracellular biological active component such as globulin or other water soluble compound of containing thereby condition is described eucaryotic cell structure.Preferred material comprises embryo, the young, new born animal or adult peg graft animal such as water flea, wheel animalcule such as arm tail wheel worm, the Artemia cysts that shells, nematode, few caterpillar, polychaete worm or insect.
Preferred embodiment on the other hand in, the size of described bioactive food compound and shape are produced to such an extent that adapt with the feed of purpose aquatic animal kind mechanism and behavioral trait.For example, in young suspension feeder, microcapsules or pearl can make by being atomised to emulsion-2 in the immersion liquid of calcium chloride that weight percentage is about 5-20% with nitrogen and collecting the gel microcapsules that size range is about 20-200mu.For the young and the back young animal that with live prey or detritus are food, microcapsules can be made into the size of about 100-1000mu.Wet screening is used to collect the microcapsules or the pearl of required size.
For the aquatic animal that feeds on insects, can make the vermiform or the bar shaped that are similar to size about 0.2mm-20mm (girth) * nematode of 0.1cm-25cm (length), few caterpillar or polychaete worm.Make bar by emulsion-2 being expressed in the calcium chloride immersion liquid that weight percentage is 5-20%.For animal such as sea urchin, snail, Bao or other is the animal of food with the water plant, can make the big or small tablet of about 0.1cm-1.0cm (thickness) * 1.0-100cm (width) * 1.0cm-100cm (length).Mould (for example stainless or plastics) that can be by utilizing suitable shape thus preparing gel makes tablet, be immersed in afterwards in the calcium chloride that weight percentage is 5-20%.
Described bioactive food compound can be preserved with semi-solid slurries in weight percentage is about calcium chloride, sodium chloride, potassium chloride or other salting liquid of 10-25% and not need drying.Can add mould inhibitor such as glycerine, propane diols or propionic acid and preserve wet product.In another aspect of this invention, under-5 ℃ or lower temperature, described bioactive food compound is carried out freezing before, with account for the about 1-50% of end article weight percentage use antifreezing agent such as methyl-sulfoxide (DMSO), glycerine, Yoshinox BHT (BHT), sucrose, gossypose, sweet mellow wine, ethylene glycol, propane diols, methyl alcohol, 1,2-propane diols, 1,3-butanediol, 2, the 3-butylene glycol.
Described bioactive food compound does not need freezing and can be kept under about 25 ℃ normal room temperature with the form of wet material; perhaps when the structural intergrity of the software invertebral living creature, embryo or other biologically active cell that need to keep lack protectiveness cell membrane or hard film, can carry out freezing preservation by antifreezing agent.
In the process of control disease, described bioactive food compound discharges probio, be used to degrade or digest the enzyme and the inhibition furan ketone compound of specific population sensitive molecule, as the part of aquatic animal feed product.Described bioactive food compound directly adds in the larya culturing groove of the storing containers of aquatile kind such as shrimp hatching place, and described bioactive food compound is eaten in this groove.Chew and when digesting described bioactive food compound, the gemma of probio will become hydrated form and sprout and be the state of activation.With the probio of dried vegetative cell form transmission in feed, intestines and will sprout sooner in ight soil and the false ight soil and begin growth.Simultaneously, described inhibition furanone will be released in the enteron aisle and will be released in the stable breeding system environments by ight soil and false ight soil subsequently.The livings bacillus of benefit begins to grow and competitive space and food and usually suppress the growth of pathogen by original position generation antibiosis under the situation of infringement pathogen.Be contained in that other bacillus strain of the present invention will produce degraded or digestion is present in the HSL in the microenvironment and the enzyme of AHL signaling molecule.Furanone is subjected to the Disease-causing gene of HSL and AHL adjusting in pathogen such as the endobacillary expression of arc with suppressing.
The acting in conjunction of probio and inhibition furanone will provide the method for the most effective control disease in environment of hatching and other aquatic environment.Described bioactive food compound provides essential micro-for juvenile prawn normal growth and survival are required and thereby a large number of nutrients has been eliminated the needs that use fresh and alive food.
Be applied to that aquaculture is cultivated and during the conventional feed in growth period, the top layer that emulsion of the present invention-1 or bioactive food compound can be contained in the aquatile feed of conventional processing such as particle be coated with wrap up in layer or dressing in.With the bioactive food compound during as the active component of top layer preparation, it is mixed in the edible oil as the top layer, in oil, form suspension, then before packing with described suspension spraying or be coated with and be rolled on the feed granules.Perhaps, directly spray or wrap up described particle, and emulsion-1 directly is used as the top layer of pellet with emulsion-1.
U.S. Pat 4,352,883 have described the suspension that forms living cells in mosanom, form immobilized cell complex compound in the immersion liquid of calcium alginate thereby then this hanging drop is added in calcium alginate, this complex compound further forms the polyelectrolyte complex compound more stable than calcium alginate complex compound with the polylysin complexing.The cell fixation of this patent and the main distinction of the present invention are:
(1) material of putting down in writing in this patent is not a two-phase emulsion system and be suspended in living cells in the hydrocolloid polymer before only relating to embedding;
(2) material of putting down in writing in this patent is except the directly edible bioactivator of animal, and what do not comprise that the present invention prepares is used to provide nutriment as the nutritional labeling of animal foodstuff or feed.
Sefton and Broughton Biochimica et Biophysica Acta, 7171,473 (1982) (encyclopedias of polymer science and engineering (Encyclopedia ofPolymer Science and Engineering), / the 9 volume, the 2nd edition (1987)) water in oil emulsion of having purchased, this emulsion has the cell that is scattered in aqueous phase and as the diethyl phthalate of oil phase.This one-level emulsion is emulsified in the mineral oil, and its mineral oil in fluid just is used for forming secondary emulsion.This process and main difference of the present invention are that the edible oil role among the present invention is that (1) is as the essential nutritional labeling of animal absorption and the source of bioactivator, (2) thus as hydrophobic coatings prevent organic substance be lost in the middle of the aquatic environment and (3) as the continuous phase of emulsion-1.
U.S. Pat 5,776,490th utilizes LWM in the hydrocolloid matrix be comprised in calcium alginate in compound, the crosslinked albumen microcapsules rather than that ionic is crosslinked and gelatin to the change of CXM notion.Emulsion-1 can be used to replace the LWM of this invention.
Embodiment
Invention has been described on the whole, can obtain further understanding of the present invention by the reference specific embodiment, and except as otherwise noted, be for illustration purpose rather than in order to limit the invention by these embodiment provided herein.
Embodiment 1
Bioactive food joint product and disease control-hatching feed
Bioactive food joint product of the present invention mixes the lipid mixtures of listing in the 2500 gram tables 1 that contains fish oil and lipophilic bioactive compound such as inhibition furanone and prepares.Then the raising mixture of listing in described lipid mixtures and the 1000 gram tables 1 that contains profitable probliotics, other bioactive compound and dry powder nutrient compounds is mixed.Thereby this mixture is fully mixed formation emulsion-1, and this emulsion is that continuous phase is that lipid mixtures and decentralized photo are the oil bag solid emulsion of forage mixture.Second kind of emulsion is to add gelatin copolymer by the mosanom that mixed emulsion-1 and 5711 grams prepare down as pH12 that table 1 is listed in to make.Thereby this mixture fully mixed forming second kind of emulsion, in second kind of emulsion, emulsion-1 is dispersed in the polymer as continuous phase.Described second kind of emulsion is atomised in the immersion liquid of 20% w/w calcium chloride solution, thereby the alginates of atomised droplets form and calcium carry out ion glue connection and forms the stable calcium-alginate matrices of a kind of physics thus.Crosslinked gel droplet filtered and recycled and form wet pulp.As shown in table 1, wet pulp utilizes the calcium chloride of 569 grams, 25% w/w or the propane diols of sodium chloride solution and 567 grams and the industrial natural gum of 11 grams to preserve, and described natural gum makes semi-solid slurries have stickiness.In prevent disease and for extra-nutrition is provided, feed the young and back young shrimp with the bioactive food joint product for preparing.Probiotic bacteria as a forage mixture part of the present invention is sprouted and growth in water and in the Gl road of shrimp, the selected bacterial strain of some of them produce antipathogen such as vibrios antibiotic and with pathogen competitive space and nutrition, and other selected bacterial strain of the present invention produces the enzyme of the specific population sensitive signal molecule of degraded or digestion pathogen.Simultaneously, the inhibition furanone that carries and transmit by the present invention suppresses the specific population sensitive procedures by the signaling molecule of pathogen such as HSL and AHL regulation and control.By using bioactive compound of the present invention and using these different mechanisms of the present invention, disease has obtained effective prevention.
The composition example of table 1. bioactive food joint product
| Biologically active lipid mixture | Gram | The bioactive feed mixture | Gram |
| Fish oil | ????2316 | Hepar siccatum | ????510.0 |
| The inhibition furanone | ????20.0 | The cuttlefish extract | ????49.0 |
| Antioxidant | ????2.5 | Fish albumen | ????20.0 |
| Lecithin | ????115.0 | Yeast extract | ????160.0 |
| Emulsifying agent | ????25.0 | Probiotic bacteria | ????150.0 |
| Cholesterol | ????10.0 | Algae | ????50.0 |
| Astaxanthin | ????9.0 | Vitamin mixtures | ????50.0 |
| Liposoluble vitamin | ????2.5 | Carotenoid pigment | ????10.0 |
| Total liquid phase | ????2500 | Trace minerals | ????1.0 |
| Total solid phase | ????1000 | ||
| Total emulsion-1 | ????3500 | ||
| Polymer | Gram | The paste additive | Gram |
| Mosanom | ????143 | Salting liquid | ????569 |
| Gelatin | ????29 | Anticorrisive agent | ????557 |
| Water | ????5539 | Natural gum | ????11 |
| Total polymer | ????5711 | Total additive | ????1137 |
| Total emulsion-2 | ????9211 |
Embodiment 2
Emulsion-1 contain as the bioactive food compound of top layer dressing by granulation and the prepared traditional animal feed of extruding, described emulsion-1 prepares by mixing the listed lipid mixtures that contains fish oil and lipophilic bioactive compound such as inhibition furanone of 2500 gram tables 2.Then the forage mixture of lipid mixtures with 1000 grams mixed, described mixture contains probiotic bacteria and/or other bioactive compound that table 2 is listed.Fully thereby the described mixture of mixture forms emulsion-1, the emulsion among this embodiment-the 1st, and oil bag solid emulsion, its continuous phase is that lipid mixtures and decentralized photo are forage mixture.Use whole emulsion by spraying or coating method with the concentration of about 1 to 300kg the oily emulsion of per metric ton animal feed then, preferably use with the concentration of about 10 to 80kg the oily emulsion of per metric ton animal feed.In this embodiment, bioactive compound of the present invention is used for traditional particle or extruding or other feed does not need second kind of emulsion, described feed has been made and can have wrapped up with emulsion-1.In the prevent disease process, adopt and raised aquatic livestock such as shrimp or fish by the conventional feed of biologically active emulsion dressing of the present invention.Sprout and growth in water and in the Gl road of shrimp as the probiotic bacteria that the part of forage mixture of the present invention is used, the selected bacterial strain of some of them produce antipathogen such as vibrios antibiotic and with pathogen competitive space and nutrition, and other selected bacterial strain of the present invention produces degraded or the specific population sensitive signal molecule of digestion pathogen such as enzyme of HSL and AHL.Simultaneously, the inhibition furanone that carries and transmit by emulsion of the present invention-1 suppresses by the signaling molecule of pathogen such as the specific population sensitive procedures of HSL and AHL regulation and control.By using bioactive compound of the present invention and using these different mechanisms of the present invention, disease has obtained effective prevention.
Table 2. is implemented as the composition of the emulsion-1 on the biologically active top layer of granulated meal
| Biologically active lipid | Gram |
| Fish oil | ????2107 |
| The inhibition furanone | ????250 |
| Antioxidant | ????3 |
| Lecithin | ????115 |
| Emulsifying agent | ????25 |
| Total liquid phase | ????2500 |
| Bioactive feed | Gram |
| Yeast extract | ????250.0 |
| Probio | ????750.0 |
| Total solid phase | ????1000 |
| Total emulsion-1 | ????3500 |
Obviously, might make multiple improvement and change to the present invention according to above-mentioned instruction.Therefore should be understood in the scope of appended claim, can implement the present invention not according to specific descriptions herein.
Claims (92)
1. control and/or prophylactic method in aquatic animal, it comprises that the lively thing feeding of feedwater contains the composition of at least a probiotic bacteria and at least a inhibition or modulating compound.
2. the process of claim 1 wherein that described probiotic bacteria and inhibition or modulating compound are integrated in the animal feed, this feed joins in the water that contains aquatic animal.
3. the process of claim 1 wherein that described probiotic bacteria and inhibition or modulating compound are integrated in the animal feed as the emulsion-1 of bioactive food compound, described emulsion-1 is as the top layer dressing of animal feed.
4. the method for claim 1, wherein said probiotic bacteria and inhibition or modulating compound be by being added in the middle of the feed that is incorporated into aquatic animal in the bioactive food compound, and described bioactive food compound is as complete feeding-stuffs and directly be added in the stable breeding system such as crawl, groove, cage, water channel, pond or other stable breeding form of aquatic animal.
5. the method for claim 1, wherein said probiotic bacteria and inhibition or modulating compound are applied to the aquatic animal feed as the emulsion-1 of bioactive food compound, and wherein said bioactive food compound is as the top layer dressing of aquatic animal feed.
6. the process of claim 1 wherein that described probiotic bacteria is selected from the bacterial isolates that includes but not limited to bacillus subtilis, bacillus laterosporus, bacillus licheniformis, bacillus azotoforlmans, Bacillus circulans, bacillus pumilus, bacillus firmus, Paenibacillus polymyxa, Paenibacillus macerans, replaces the zygosaccharomyces bacterium.
7. the process of claim 1 wherein that described inhibition compound is furanone or other lipophilic bioactive compound.
8. the process of claim 1 wherein that controlled pathogen is the Gram-negative bacteria that includes but not limited to vibrio harveyi, vibrio parahaemolytious, Vibrio splindidus, vibrio mimicus, comma bacillus, separates alginic acid vibrios, Vibrio anguillarum, other vibrio bacterium, Aeromonas bacterium.
9. the process of claim 1 wherein that controlled pathogen is the gram-positive bacteria that includes but not limited to streptococcus, core bar bacterium, micrococcus luteus etc.
10. the process of claim 1 wherein that control of pathogens occurs in the animal alimentary canal 9.
11. the method for claim 1, wherein control of pathogens occurs in the environment of the animal that comprises feed vessel, feed dish, fence, matting, crawl, groove, cage, water channel, pond, and in water, surface and the deposit of these environment and other stable breeding system.
12. prepare the method for bioactive food compound, comprising:
Prepare first emulsion (emulsion-1), this first emulsion contains the emulsion of oil bag solid or solid bag oil, this emulsion is made up of with the lipophilic bioactive compound that is dissolved in the edible oil that forms oil phase bioactivator that forms solid phase and powdery nutrients, and prepare second kind of emulsion, this second kind of emulsion contains the emulsion of polymer bag oil or polymer bag solid, wherein decentralized photo contains emulsion-1 and hydrocolloid polymer as continuous phase, and
Make described hydrocolloid polymer be exposed to ion, thereby the described polymer of ionomer and form physics stabilizing gel matrix is embedded in emulsion-1 in second kind of emulsion, thereby make bioactive composite.
13. the method for claim 12, wherein the oil phase of first kind of emulsion is selected from fish oil, refined fish oil and vegetable oil and contains lipophilic bioactive compound.
14. the method for claim 13, wherein said lipophilic bioactive compound are that inhibition furanone (anfuranone) and described grease separation are from herring oil, salmon oil, long tail anchovy oil, pilchard oil, tunny fish oil, mackerel oil, thin squama cucumber fish oil, Squid Oil, pollack fish oil, cod oil, edible fish oil additive, soybean oil, safflower oil, corn oil and palm-kernel oil.
15. the method for claim 12, wherein the continuous phase polymer of second kind of emulsion is made up of the polymer that weight percentage is about 0.5-4.0%, and described polymer is selected from alginates and carragheen polymer.
16. the method for claim 12, wherein the continuous phase polymer of second kind of emulsion makes in deionized water by adding at least a ion that is selected from sodium, potassium and calcium ion in 45-85 ℃.
17. the method for claim 12, wherein said polymer mixes with the composition that is selected from gelatin, zein, polylysin, poly arginine, shitosan, Arabic gum and tracasol goods with the weight percentage of about 0.1-3.5% in water.
18. the method for claim 12, the part by weight of wherein said solid and oil is between about 0.01: 1 to 100: 1, preferably between 0.1: 1 to 3: 1.
19. the method for claim 13 has wherein added at least a material that is selected from lecithin, cholesterol and emulsifying agent, antioxidant, liposoluble vitamin and astaxanthin in oil.
20. the method for claim 13, wherein said oil contains mixture, and this mixture contains edible oil, weight percentage, and to be about 0.0001 to 50% lipophilic bioactive compound such as inhibition furanone, weight percentage be that 0.5 to 5% emulsifying agent, weight percentage are about 1 to 10% lecithin, weight percentage and are about 1 to 10% cholesterol and weight percentage and are about 0.01 to 0.5% antioxidant.
21. the method for claim 20, wherein said oil contain every kilogram of about vitamin A of 2000 to 8000IU, about vitamin D of 1000 to 4000IU
3, about vitamin E of 2000 to 8000IU, about vitamin K of 20 to 80mg and about 2000mg astaxanthin.
22. the method for claim 13, wherein said oil contain weight percentage greater than 1% omega-fatty acid, weight percentage greater than 0.1% eicosapentaenoic acid and weight percentage greater than 0.1% DHA.
23. the method for claim 13 wherein is heated to 25 to 40 ℃ with described oil before adding described bioactivator and powdery nutrients.
24. the method for claim 12, wherein said bioactivator and powdery nutrients are to be selected from least a in bacterial spore, bacteria cell wall, bacterial cell, yeast cell wall, yeast extract, food yeast, brewer's yeast, yeast cells and the alga cells.
25. the method for claim 24, wherein said bacterial cell are at least a bacteriums that is selected from bacillus, Bacillus acidi lactici, streptococcus, Bifidobacterium, leukonid and replaces monad.
26. the method for claim 12, wherein said oil phase before adding powdery nutrients and bioactivator, contain total concentration greater than the omega-fatty acid of about 1% weight percentage, total concentration greater than the eicosapentaenoic acid (EPA) of about 0.1% weight percentage and total concentration DHA (DHA) greater than about 0.1% weight percentage.
27. the method for claim 24, wherein said alga cells are at least a alga cells that includes but not limited to flagellate, Schizochytrium, Du Shi algae, Chaetoceros, Tetraselmis, Skeletonema Greville, Nannochloropsis, hailian seaweed, brown algae, Isochrysis galbana, Pavlova that is selected from
28. the method for claim 12, the solid phase of wherein said first kind of emulsion contain the solid matter that is selected from bacterial components, yeast material, algae substances, pharmaceutical substance, animal protein product, plant protein products and birds, beasts and eggs product.
29. the method for claim 12, the solid phase of wherein said first kind of emulsion contains the solid matter that is selected from bacterial components, yeast material, algae substances, medicament, mash feed, animal protein product, plant protein products, birds, beasts and eggs product, cobalamin, D-biotin, D-pantothenic acid, folic acid, L-ascorbic acid-2-polyphosphoric acid fat, inositol, nicotinic acid, p-aminobenzoic acid, pyridoxine hydrochloride, riboflavin, thiamine chloride and Choline Chloride, and wherein said bacterial components is selected from bacterial spore, bacteria cell wall and bacterial cell; Described yeast material is selected from yeast cell wall, yeast extract, food yeast, brewer's yeast, yeast cells.
30. the method for claim 29, the solid phase of wherein said first kind of emulsion contain the bacterial components that is selected from Bacillus acidi lactici, bacillus, streptococcus, Bifidobacterium, leukonid and replaces monad.
31. the method for claim 30, wherein said bacterial components exists with the weight percentage of the 0.1-95% that accounts for solid phase.
32. the method for claim 28, the solid phase of wherein said first kind of emulsion contain the yeast material that is selected from torula and Phaffia that weight percentage is about 0.1-50%.
33. the method for claim 28, wherein said yeast material exists with the weight percentage of the 0.1-50% that accounts for solid phase.
34. the method for claim 28, the solid phase of wherein said first kind of emulsion contain the algae substances that is selected from flagellate, Schizochytrium, Du Shi algae, Chaetoceros, Tetraselmis, Skeletonema Greville, Nannochloropsis, hailian seaweed, brown algae, Isochrysis galbana and Pavlova.
35. the method for claim 34, wherein said yeast material exists with the weight percentage of the 1-80% that accounts for solid phase.
36. containing, the method for claim 31, the solid phase of wherein said first kind of emulsion be selected from Sarafin, the antibiotic of Repirinast and terramycin.
37. the method for claim 36, wherein said antibiotic exists with the weight percentage of the 0.01-50% that accounts for solid phase.
38. the method for claim 31, the solid phase of wherein said first kind of emulsion contains the animal protein product with the weight percentage of the 0-95% that accounts for solid phase.
39. the method for claim 31, the solid phase of wherein said first kind of emulsion contains plant protein products with the weight percentage of the 0-95% that accounts for solid phase.
40. the method for claim 31, the solid phase of wherein said first kind of emulsion contains the birds, beasts and eggs product with the weight percentage of the 0-25% that accounts for solid phase.
41. the method for claim 29, the solid phase of wherein said first kind of emulsion contains the cobalamin of the 40-60mg/kg that has an appointment; The D-biotin of about 5-20mg/kg; The D-pantothenic acid of about 250-350mg/kg; The folic acid of about 10-30mg/kg; The L-ascorbic acid of about 1000-4000mg/kg-2-polyphosphoric acid fat (STAY-C, ascorbic stable form); The inositol of about 3000-4000mg/kg; The nicotinic acid of about 600-800mg/kg; The p-aminobenzoic acid of about 350-450mg/kg; The pyridoxine hydrochloride of about 40-60mg/kg; The riboflavin of about 125-175mg/kg; The thiamine chloride of about 50-80mg/kg and the Choline Chloride of about 6500-7500mg/kg.
42. the method for claim 12 wherein is embedded in more frangible biological active component in second kind of emulsion.
43. the method for claim 42, wherein said more frangible biological active component is selected from invertebral living creature embryo and invertebral living creature.
44. the method for claim 42, wherein said more frangible biological active component are selected from the metazoa of metazoa alive, freezing metazoa, freeze-drying, the protozoan that lives, freezing protozoan, the protozoan of freeze-drying, the plant that lives or animal tissue, freezing plant or the plant or the animal tissue of animal tissue and freeze-drying.
45. the method for claim 42, wherein said more frangible biological active component is selected from an embryo of angular motion thing, the young, neonate and adult body.
46. the method for claim 41, wherein said more frangible biological active component are selected from water flea, wheel animalcule, the Artemia cysts that shells, nematode, few caterpillar, polychaete worm and insect.
47. the method for claim 46, wherein said wheel animalcule are arm tail wheel worms.
48. be used for the bioactive food compound of raising aquatic animals, be grouped into by following one-tenth: first kind of emulsion (emulsion-1), be the emulsion of oil bag solid or solid bag oil, its bioactivator and powdery nutrients form solid phase and lipophilic bioactive compound is dissolved in the edible oil that forms oil phase; And second kind of emulsion, form by the emulsion of polymer bag oil or polymer bag solid, wherein form decentralized photo by emulsion-1 and hydrocolloid polymer as continuous phase; This compound is exposed to ion, thereby makes the hydrocolloid polymer ionomer form physics stabilizing gel matrix, emulsion-1 is embedded in second kind of emulsion, thereby constitutes bioactive composite.
49. the bioactive food compound of claim 48, the continuous phase polymer of wherein said second emulsion is made up of the polymer that weight percentage is about 0.5-4.0%, and this polymer is selected from that alginates and carragheen polymer and wherein said polymer are selected from the ion of sodium, potassium and calcium by interpolation and is crosslinked.
50. the bioactive food compound of claim 48, the oil phase of wherein said first kind of emulsion is selected from fish oil, refined fish oil and vegetable oil.
51. the bioactive food compound of claim 48, wherein said oil phase are selected from herring oil, salmon oil, long tail anchovy oil, pilchard oil, tunny fish oil, mackerel oil, thin squama cucumber fish oil, Squid Oil, pollack fish oil, cod oil, edible fish oil additive, soybean oil, safflower oil, corn oil and palm-kernel oil.
52. the bioactive food compound of claim 48, wherein said solid and oil part by weight between about 0.01: 1 to 100: 1, preferred 0.1: 1 to 3: 1.
53. the bioactive food compound of claim 52 wherein adds at least a material that is selected from lecithin, cholesterol, emulsifying agent, antioxidant, liposoluble vitamin and astaxanthin in oil.
54. the bioactive food compound of claim 48, the mixture that wherein constitutes described oil comprise that edible oil, weight percentage are about 0.5 to 5% emulsifying agent, weight percentage and are about 1 to 10% lecithin, weight percentage and are about 1 to 10% cholesterol and weight percentage and are about 0.01 to 0.5% antioxidant.
55. the bioactive food compound of claim 48, wherein said oil contain every kilogram of about vitamin A of 2000 to 8000IU, about vitamin D of 1000 to 4000IU
3, about vitamin E of 2000 to 8000IU, about vitamin K of 20 to 80mg and about 2000mg astaxanthin.
56. the bioactive food compound of claim 48, wherein said oil contain weight percentage greater than 1% omega-fatty acid, weight percentage greater than 0.1% eicosapentaenoic acid and weight percentage greater than 0.1% DHA.
57. the bioactive food compound of claim 48 wherein is heated to 25 to 40 ℃ with described oil before adding described bioactivator and powdery nutrients.
58. the bioactive food compound of claim 48, wherein said bioactivator are at least a materials that is selected from bacterial spore, bacteria cell wall, bacterial cell, yeast cell wall, yeast extract, food yeast, brewer's yeast, yeast cells and alga cells.
59. the bioactive food compound of claim 58, wherein said bacterial cell contain at least a bacterium that is selected from Bacillus acidi lactici, bacillus, streptococcus, Bifidobacterium, leukonid and replaces monad.
60. the bioactive food compound of claim 58, wherein said bioactivator contain total concentration greater than the omega-fatty acid of about 1% weight percentage, total concentration greater than the eicosapentaenoic acid (EPA) of about 0.1% weight percentage and total concentration DHA (DHA) greater than about 0.1% weight percentage.
61. the bioactive food compound of claim 58, wherein said alga cells contain at least a alga cells that includes but not limited to flagellate, Schizochytrium, Du Shi algae, Chaetoceros, Tetraselmis, Skeletonema Greville, Nannochloropsis, hailian seaweed, brown algae, Isochrysis galbana, Pavlova that is selected from
62. the bioactive food compound of claim 48, the solid phase of wherein said first kind of emulsion contain the solid matter that is selected from bacterial components, yeast material, algae substances, pharmaceutical substance, animal protein product, plant protein products and birds, beasts and eggs product
63. the bioactive food compound of claim 48, the solid phase of wherein said first kind of emulsion contains the solid matter that is selected from bacterial components, yeast material, algae substances, medicament, mash feed, and wherein said bacterial components is selected from bacterial spore, bacteria cell wall and bacterial cell; Described yeast material is selected from yeast cell wall, yeast extract, food yeast, brewer's yeast, yeast cells; Described algae substances is selected from the alga cells goods; Described mash feed is selected from animal protein product, plant protein products, birds, beasts and eggs product, cobalamin, D-biotin, D-pantothenic acid, folic acid, L-ascorbic acid-2-polyphosphoric acid fat, inositol, nicotinic acid, p-aminobenzoic acid, pyridoxine hydrochloride, riboflavin, beclotiamine and Choline Chloride.
64. the bioactive food compound of claim 48, the solid phase of wherein said first kind of emulsion contain the bacterial components that is selected from Bacillus acidi lactici, bacillus, streptococcus, Bifidobacterium, leukonid and replaces monad.
65. the bioactive food compound of claim 64, wherein said bacterial components exists with the weight percentage of the 0.1-95% that accounts for solid phase.
66. the bioactive food compound of claim 48, the solid phase of wherein said first kind of emulsion contain the yeast material that is selected from torula and Phaffia that weight percentage is about 0.1-50%.
67. the bioactive food compound of claim 66, wherein said yeast material exists with the weight percentage of the 0.1-50% that accounts for solid phase.
68. the bioactive food compound of claim 48, the solid phase of wherein said first kind of emulsion contain the algae substances that is selected from flagellate, Schizochytrium, Du Shi algae, Chaetoceros, Tetraselmis, Skeletonema Greville, Nannochloropsis, hailian seaweed, brown algae, Isochrysis galbana and Pavlova.
69. the bioactive food compound of claim 67, wherein said weight percentage with the 1-80% that accounts for solid phase exists.
70. containing, the bioactive food compound of claim 68, the solid phase of wherein said first kind of emulsion be selected from Sarafin, the antibiotic of Repirinast and terramycin.
71. the bioactive food compound of claim 69, wherein said antibiotic exists with the weight percentage of the 0.01-50% that accounts for solid phase.
72. the bioactive food compound of claim 62, the solid phase of wherein said first kind of emulsion contains the animal protein product with the weight percentage of the 0-95% that accounts for solid phase.
73. the bioactive food compound of claim 62, the solid phase of wherein said first kind of emulsion contains plant protein products with the weight percentage of the 0-95% that accounts for solid phase.
74. the bioactive food compound of claim 62, the solid phase of wherein said first kind of emulsion contains the birds, beasts and eggs product with the weight percentage of the 0-25% that accounts for solid phase.
75. the bioactive food compound of claim 62, the solid phase of wherein said first kind of emulsion contains the Cobastab of the 40-60mg/kg that has an appointment
12The D-biotin of about 5-20mg/kg; The D-pantothenic acid of about 250-350mg/kg; The folic acid of about 10-30mg/kg; The L-ascorbic acid of about 1000-4000mg/kg-2-polyphosphoric acid fat (STAY-C, ascorbic stable form); The inositol of about 3000-4000mg/kg; The nicotinic acid of about 600-800mg/kg; The p-aminobenzoic acid of about 350-450mg/kg; The pyridoxine hydrochloride of about 40-60mg/kg; The riboflavin of about 125-175mg/kg; The beclotiamine of about 50-80mg/kg and the Choline Chloride of about 6500-7500mg/kg.
76. the bioactive food compound of claim 48 wherein is embedded in more frangible biological active component in second kind of emulsion.
77. the bioactive food compound of claim 76, wherein said more frangible biological active component is selected from invertebral living creature embryo and invertebral living creature.
78. the described bioactive food compound of claim 76, wherein said more frangible biological active component are selected from the metazoa of metazoa alive, freezing metazoa, freeze-drying, the protozoan that lives, freezing protozoan, the protozoan of freeze-drying, the plant that lives or animal tissue, freezing animal or plant tissue and the plant or the animal tissue of freeze-drying.
79. the bioactive food compound of claim 76, wherein said more frangible biological active component is selected from embryo, the young, neonate and the adult body of peg graft animal.
80. the bioactive food compound of claim 76, wherein said more frangible biological active component are selected from a kind of in water flea, wheel animalcule, the Artemia cysts that shells, nematode, few caterpillar, polychaete worm and the insect.
81. the bioactive food compound of claim 80, wherein said wheel animalcule are arm tail wheel worms.
82. be used for the bioactive food compound of raising aquatic animals, this compound contains: first kind of emulsion (emulsion-1), emulsion for oil bag solid or solid bag oil, its bioactivator and powdery nutrients form solid phase and edible oil formation oil phase, described grease separation is from fish oil, refined fish oil and vegetable oil, described solid and oil part by weight between about 0.01: 1 to 100: 1, preferred 0.1: 1 to 3: 1; With second kind of emulsion, contain the emulsion-l that is dispersed in the hydrocolloid, described emulsion-1 is dispersed in described second kind of emulsion, described hydrocolloid polymer is as continuous phase, described hydrocolloid polymer ionomer forms the stable matrix of physics, and described hydrocolloid continuous phase comprises the invertebral living creature of dispersion.
83. the described bioactive food compound of claim 82, wherein said oil phase are selected from herring oil, salmon oil, long tail anchovy oil, pilchard oil, tunny fish oil, mackerel oil, thin squama cucumber fish oil, Squid Oil, pollack fish oil, cod oil, edible fish oil additive, soybean oil, safflower oil, corn oil and palm-kernel oil.
84. the bioactive food compound of claim 83 wherein adds at least a material that is selected from lecithin, cholesterol, emulsifying agent, antioxidant, liposoluble vitamin and astaxanthin in oil.
85. the bioactive food compound of claim 82, the mixture that wherein constitutes described oil contain edible oil, weight percentage and are about 0.0001 to 50% lipophilic bioactive compound such as inhibition furanone, weight percentage and are about 0.5 to 5% emulsifying agent, weight percentage and are about 1 to 10% lecithin, weight percentage and are about 1 to 10% cholesterol and weight percentage and are about 0.01 to 0.5% antioxidant.
86. the bioactive food compound of claim 82, wherein said bioactivator contain at least a material that is selected from bacterial spore, bacteria cell wall, bacterial cell, yeast cell wall, yeast extract, food yeast, brewer's yeast, yeast cells and alga cells.
87. the bioactive food compound of claim 86, wherein said bacterial cell contain at least a bacterium that is selected from Bacillus acidi lactici, bacillus, streptococcus, Bifidobacterium, leukonid and replaces monad.
88. the bioactive food compound of claim 86, wherein said bioactivator contain total concentration greater than the omega-fatty acid of about 1% weight percentage, total concentration greater than the eicosapentaenoic acid (EPA) of about 0.1% weight percentage and total concentration DHA (DHA) greater than about 0.1% weight percentage.
89. the bioactive food compound of claim 86, wherein said alga cells contain at least a alga cells that includes but not limited to flagellate, Schizochytrium, Du Shi algae, Chaetoceros, Tetraselmis, Skeletonema Greville, Nannochloropsis, hailian seaweed, brown algae, Isochrysis galbana, Pavlova that is selected from.
90. the bioactive food compound of claim 82, the solid phase of wherein said first kind of emulsion contain the solid matter that is selected from bacterial components, yeast material, algae substances, pharmaceutical substance, animal protein product, plant protein products and birds, beasts and eggs product.
91. the bioactive food compound of claim 82, the solid phase of wherein said first kind of emulsion contains the solid matter that is selected from bacterial components, yeast material, algae substances, medicament, mash feed, and wherein said bacterial components is selected from bacterial spore, bacteria cell wall and bacterial cell; Described yeast material is selected from yeast cell wall, yeast extract, food yeast, brewer's yeast, yeast cells; Described algae substances is selected from the alga cells goods; Described mash feed is selected from animal protein product, plant protein products, birds, beasts and eggs product, cobalamin, D-biotin, D-pantothenic acid, folic acid, L-ascorbic acid-2-polyphosphoric acid fat, inositol, nicotinic acid, p-aminobenzoic acid, pyridoxine hydrochloride, riboflavin, thiamine chloride and Choline Chloride.
92. the bioactive food compound of claim 91, wherein hydrocolloid continuous phase further contains the invertebral living creature of dispersion.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US21353800P | 2000-06-23 | 2000-06-23 | |
| US60/213,538 | 2000-06-23 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1454058A true CN1454058A (en) | 2003-11-05 |
| CN100456949C CN100456949C (en) | 2009-02-04 |
Family
ID=22795483
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB018128130A Expired - Lifetime CN100456949C (en) | 2000-06-23 | 2001-06-22 | Bioactive food complex, method for making bioactive food complex product and method for controlling disease |
Country Status (5)
| Country | Link |
|---|---|
| CN (1) | CN100456949C (en) |
| AU (1) | AU2001268078A1 (en) |
| EC (1) | ECSP024398A (en) |
| MX (1) | MXPA02012825A (en) |
| WO (1) | WO2002000035A1 (en) |
Cited By (5)
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| CN101953429A (en) * | 2009-07-21 | 2011-01-26 | 天津滨海索尔特生物技术中心有限公司 | Marine fish fortification microcapsule aqueous solution agent and preparation method thereof |
| CN103734520A (en) * | 2013-12-20 | 2014-04-23 | 大连赛姆生物工程技术有限公司 | Self-ingestion type plant source composite pharmaceutical effect nutriment for horse and preparation method of self-ingestion type plant source composite pharmaceutical effect nutriment |
| CN104144607A (en) * | 2012-02-21 | 2014-11-12 | 先进生物营养公司 | Compositions and methods for target delivering a bioactive agent to aquatic organisms |
| CN104144607B (en) * | 2012-02-21 | 2016-11-30 | 先进生物营养公司 | For by the compositions of bioactivator targeted delivery to aquatic organism and method |
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| WO2002052949A1 (en) * | 2001-01-08 | 2002-07-11 | Gormar Marketing Limited | Autoinducer compounds and their uses |
| JP4463109B2 (en) * | 2002-11-07 | 2010-05-12 | アドバンスド バイオニュートリション コーポレーション | Dietary supplement and aquatic animal feeding method |
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| GB0313268D0 (en) | 2002-12-10 | 2003-07-16 | Smith Anthony G | Fish feed |
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| US8802171B2 (en) | 2004-05-25 | 2014-08-12 | James B. Watson | Live organism product |
| US20120114786A1 (en) * | 2004-05-25 | 2012-05-10 | Watson James B | Live bacteria product |
| US20050266027A1 (en) * | 2004-05-25 | 2005-12-01 | Watson James B | Live organism product |
| DE102004025869A1 (en) * | 2004-05-27 | 2005-12-22 | Süd-Chemie AG | Bioprotektivum |
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| US5698246A (en) * | 1996-01-29 | 1997-12-16 | Cargill, Incorporated | Foodstuff for and method of feeding crustaceans and fish |
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2001
- 2001-06-22 AU AU2001268078A patent/AU2001268078A1/en not_active Abandoned
- 2001-06-22 CN CNB018128130A patent/CN100456949C/en not_active Expired - Lifetime
- 2001-06-22 MX MXPA02012825A patent/MXPA02012825A/en active IP Right Grant
- 2001-06-22 WO PCT/US2001/016489 patent/WO2002000035A1/en active Application Filing
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2002
- 2002-12-19 EC EC2002004398A patent/ECSP024398A/en unknown
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| CN103734520B (en) * | 2013-12-20 | 2015-07-22 | 大连赛姆生物工程技术有限公司 | Self-ingestion type plant source composite pharmaceutical effect nutriment for horse and preparation method of self-ingestion type plant source composite pharmaceutical effect nutriment |
| CN112601453A (en) * | 2018-08-29 | 2021-04-02 | Nch公司 | Systems, methods, and compositions for incubating spores for use in aquaculture, agriculture, wastewater, and environmental remediation applications |
Also Published As
| Publication number | Publication date |
|---|---|
| CN100456949C (en) | 2009-02-04 |
| MXPA02012825A (en) | 2004-07-30 |
| AU2001268078A1 (en) | 2002-01-08 |
| ECSP024398A (en) | 2003-06-25 |
| WO2002000035A1 (en) | 2002-01-03 |
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