CN105166574A - Microencapsulated feed for resisting vibriosis of prawn larvae - Google Patents
Microencapsulated feed for resisting vibriosis of prawn larvae Download PDFInfo
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- CN105166574A CN105166574A CN201510667819.8A CN201510667819A CN105166574A CN 105166574 A CN105166574 A CN 105166574A CN 201510667819 A CN201510667819 A CN 201510667819A CN 105166574 A CN105166574 A CN 105166574A
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Abstract
The invention discloses microencapsulated feed for resisting vibriosis of prawn larvae. The microencapsulated feed is prepared from three kinds of raw materials, namely, prawn larvae vibriosis-resisting egg yolk antibody lyophilized powder, astragalus powder and beta-cyclodextrin, wherein the prawn larvae vibriosis-resisting egg yolk antibody lyophilized powder is mainly used for reducing or preventing the infection of the prawn larvae from vibrio parahaemolyticus and vibrio harveyi; the astragalus powder is used for increasing the immunity of the prawn larvae; the beta-cyclodextrin is mainly used for enabling the prawn larvae vibriosis-resisting egg yolk antibody lyophilized powder to achieve embedding and slow releasing functions. According to the microencapsulated feed disclosed by the invention, 80 to 85 percent of the prawn larvae vibriosis-resisting egg yolk antibody lyophilized powder of which the antibody titer is 210, 10 to 12 percent of the astragalus and 5 to 8 percent of beta-cyclodextrin are prepared into microencapsulated dry powder through spray drying, the microencapsulated feed is fed and used together with prawn slices sold in the market according to a certain ratio, the diseases caused by the vibrio parahaemolyticus and vibrio harveyi during a prawn larvae cultivation period can be prevented and treated, the prawn nursery yield can be increased, and the economic benefit of prawn nursery production can be remarkably increased; the problem that the vibriosis is easy to cause during a prawn nursery production process at present can be solved.
Description
Technical field
The present invention relates to a kind of micro-encapsulated diet used for aquiculture, particularly a kind of micro-encapsulated diet for anti-prawn larvae vibriosis.
Background technology
Prawn culturing is the pillar industry in China coast fishery economic, and in batch production Seedling production, the bacteriosis incidence of disease is the highest, it is maximum to endanger, generally from morbidity to the young, death only needs tens hours, pond young cumulative mortality of falling ill in 1-2 days time serious reaches more than 90%, and young symptom shows as vigor and obviously declines, swimming torpescence, phototaxis is weak, belly arches, and turns round and round in water, does not ingest, body colour bleaches, and its cause of disease is mainly vibrios.Of a great variety due to pathogenic vibrio during prawn seed-rearing, and infection type is complicated, so be difficult to accomplish the effectively prevention and control of pointed ground in nursery is produced, with the major obstacles causing these diseases to be formed restriction aquaculture development, produce to nursery and cause very large economic loss.Therefore, the efficient prevention and control of vibriosis penaeus, to reduce its extent of injury to greatest extent, can improve the economic benefit that prawn seed-rearing is produced significantly.
The Occurrence & epidemic of current prawn seed-rearing disease that very difficult control vibrio parahaemolytious and vibrio harveyi cause in producing, market lacks the anti-vibriosis feed that a kind of applicable prawn larvae easy to use uses.Therefore, develop prawn larvae anti-vibriosis micro-encapsulated diet, for the Occurrence & epidemic effectively controlling vibriosis in prawn seed-rearing production, there is important application prospect.
Summary of the invention
Technical problem to be solved by this invention is for the deficiencies in the prior art, provide a kind of and improve prawn larvae immunity, reduce during prawn seed-rearing is produced caused by cause of disease vibrio parahaemolytious and vibrio harveyi harm, to meet the micro-encapsulated diet of disease-resistant demand that prawn seed-rearing is produced; And this micro-encapsulated diet can directly be thrown something and fed in prawn seed-rearing is produced.
Technical problem to be solved by this invention is realized by following technical scheme.The present invention is a kind of anti-prawn larvae vibriosis micro-encapsulated diet, is characterized in, it forms micro-encapsulated diet by three kinds of raw materials of following weight proportion,
(1) anti-prawn larvae vibriosis Yolk antibody freeze-dried powder 80 ~ 85%;
(2) astragalus membranaceus powder 10% ~ 12%;
(3) beta-schardinger dextrin-5 ~ 8%;
Described anti-prawn larvae vibriosis Yolk antibody method for preparing freeze-dried powder step is as follows:
1. by cause prawn larvae vibriosis cause of disease vibrio parahaemolytious (
vibrioparahaemolyticus) VP1 and vibrio harveyi (
vibrioharveyi) kx1 bacterial strain is seeded on ordinary nutritional broth bouillon respectively, 24h cultivated by 28-30 DEG C of shaking table, adds formaldehyde 28 DEG C of deactivation 48h of volume final concentration 0.3%, centrifugal deactivation thalline;
2. decoct after getting Radix Astragali distilled water immersion 30-60min, filter, being concentrated into final concentration is former medicine 1g/mL, obtains Radix Astragali leachate;
3. mixed with Radix Astragali leachate by two kinds of deactivation thalline, obtained water aqua type vaccine, in vaccine liquid, cause of disease vibrio parahaemolytious VP1 and vibrio harveyi kx1 immunogene are respectively 10
11cells/mL, Radix Astragali leachate is 1g/mL;
4. with two kinds of obtained pathogenic vibrio water aqua type inactivated vaccines, adopt the adult laying hen of method immunity that is oral and collunarium combined immunization, the 1st time immunizing dose is: oral immunity joins in drinking-water by vaccine liquid, and addition is 5mL; Nasal immunization is that vaccine drop is entered laying hen nasal cavity, and infusion volume is 1mL; Press preceding method booster immunization, continuous booster immunization 5 times after one week, altogether immunity 6 times, immunity terminates latter 6th week, and Yolk antibody is tired and reached 2
10or more; To collect produce by immune chicken egg, isolate yolk, through freeze drying, obtained anti-prawn larvae vibriosis Yolk antibody freeze-dried powder.
Involvedly in the present invention produce in anti-prawn larvae vibriosis Yolk antibody freeze-dried powder method; with two kinds of obtained pathogenic vibrio water aqua type inactivated vaccines; adopt the adult laying hen of method immunity that is oral and collunarium combined immunization; it is not the immunity realizing chicken self; therefore it does not belong to methods for the treatment of, but disposes animal to obtain its egg that produces by the method for non-surgery operation, and then isolates the yolk realizing the object of the invention; therefore, the method belongs to the scope of patent protection.
Anti-prawn larvae vibriosis micro-encapsulated diet of the present invention, further preferred technical side is: anti-prawn larvae vibriosis Yolk antibody method for preparing freeze-dried powder step 4. in, during freeze drying, egg yolk liquid thickness 6mm, freeze-drying time is 10h.
Anti-prawn larvae vibriosis micro-encapsulated diet of the present invention, preferred technical side is further: the particle diameter of described astragalus membranaceus powder is at 800 ~ 1000 orders, and the purity of described beta-schardinger dextrin-is not less than 80%.
Anti-prawn larvae vibriosis micro-encapsulated diet of the present invention, preferred technical side is further: adopt spray-dired mode, obtained microcapsules dry powder feeding.
Anti-prawn larvae vibriosis micro-encapsulated diet of the present invention, preferred technical side is further: the weight proportion of each raw material is preferably as follows:
(1) anti-prawn larvae vibriosis Yolk antibody freeze-dried powder 82.5%;
(2) astragalus membranaceus powder 11%;
(3) beta-schardinger dextrin-6.5%.
Prawn larvae cause of disease vibrio parahaemolytious VP1 bacterial strain involved in the present invention is at open source literature " Isolationandcharacterizationof
vibrioparahaemolyticusand
vibriorotiferianusassociatedwithmassmortalitiesofChineseshrimp (
fenneropenaeuschinensis) " in disclosed (bacterial strain VP1), above-mentioned open source literature is published in " JournalofShellfishResearch ", 2014,33(1): 61-68.Prawn larvae cause of disease vibrio harveyi kx1 bacterial strain of the present invention is in open source literature " qualification of Chinese prawn mysis cause of disease vibrio harveyi and virulence gene detect " disclosed (bacterial strain kx1), above-mentioned open source literature is published in " fishery science progress ", 2014,35(3): 105-111.DNA sequence dna length and the Genebank accession number of bacterial strain VP1 and kx1 see the following form:
Bacterial strain VP1 and kx1 is public material, is now kept at Huaihai Institute of Technology oceanography institute laboratory, rises in Two decades years in present patent application day, if the public needs, Huaihai Institute of Technology oceanography institute can externally provide in laboratory.
The using method of a kind of anti-prawn larvae vibriosis micro-encapsulated diet of the present invention is as follows: the prawn larvae nurturing period, by prawn each phase young of directly throwing something and feeding after different mesh bolting silk, also can throw something and feed together with commercially available young prawn slice by the ratio of 30% and use.Anti-prawn larvae vibriosis micro-encapsulated diet, not only has good resistant effect, and easy to use, has again higher nutritive value simultaneously.
The present invention's anti-prawn larvae vibriosis micro-encapsulated diet, the resistance of prawn larvae to cause of disease vibrio parahaemolytious and vibrio harveyi can be strengthened, improve the survival rate of prawn larvae, improve the economic benefit that nursery is produced, to solve the problem that vibriosis easily occurs in current prawn seed-rearing production process.
Detailed description of the invention
Technical problem to be solved by this invention is realized by following technical scheme, so that those skilled in the art understands the present invention further, and do not form the restriction to its right.
Embodiment 1, a kind of anti-prawn larvae vibriosis micro-encapsulated diet, it forms micro-encapsulated diet by three kinds of raw materials of following weight proportion,
(1) anti-prawn larvae vibriosis Yolk antibody freeze-dried powder 80%;
(2) astragalus membranaceus powder 12%;
(3) beta-schardinger dextrin-8%;
Described anti-prawn larvae vibriosis Yolk antibody method for preparing freeze-dried powder step is as follows:
1. by cause prawn larvae vibriosis cause of disease vibrio parahaemolytious (
vibrioparahaemolyticus) VP1 and vibrio harveyi (
vibrioharveyi) kx1 bacterial strain is seeded on ordinary nutritional broth bouillon respectively, 24h cultivated by 28-30 DEG C of shaking table, adds formaldehyde 28 DEG C of deactivation 48h of volume final concentration 0.3%, centrifugal deactivation thalline;
2. decoct after getting Radix Astragali distilled water immersion 30-60min, filter, being concentrated into final concentration is former medicine 1g/mL, obtains Radix Astragali leachate;
3. mixed with Radix Astragali leachate by two kinds of deactivation thalline, obtained water aqua type vaccine, in vaccine liquid, cause of disease vibrio parahaemolytious VP1 and vibrio harveyi kx1 immunogene are respectively 10
11cells/mL, Radix Astragali leachate is 1g/mL;
4. with two kinds of obtained pathogenic vibrio water aqua type inactivated vaccines, adopt the adult laying hen of method immunity that is oral and collunarium combined immunization, the 1st time immunizing dose is: oral immunity joins in drinking-water by vaccine liquid, and addition is 5mL; Nasal immunization is that vaccine drop is entered laying hen nasal cavity, and infusion volume is 1mL; Press preceding method booster immunization, continuous booster immunization 5 times after one week, altogether immunity 6 times, immunity terminates latter 6th week, and Yolk antibody is tired and reached 2
10or more; To collect produce by immune chicken egg, isolate yolk, through freeze drying, obtained anti-prawn larvae vibriosis Yolk antibody freeze-dried powder.
Embodiment 2, a kind of anti-prawn larvae vibriosis micro-encapsulated diet, it forms micro-encapsulated diet by three kinds of raw materials of following weight proportion,
(1) anti-prawn larvae vibriosis Yolk antibody freeze-dried powder 85%;
(2) astragalus membranaceus powder 10%%;
(3) beta-schardinger dextrin-5%;
Described anti-prawn larvae vibriosis Yolk antibody method for preparing freeze-dried powder is identical with embodiment 1.
Embodiment 3, a kind of anti-prawn larvae vibriosis micro-encapsulated diet, it forms micro-encapsulated diet by three kinds of raw materials of following weight proportion,
(1) anti-prawn larvae vibriosis Yolk antibody freeze-dried powder 82.5%;
(2) astragalus membranaceus powder 11%;
(3) beta-schardinger dextrin-6.5%;
Described anti-prawn larvae vibriosis Yolk antibody method for preparing freeze-dried powder is identical with embodiment 1.
Embodiment 4, in the anti-prawn larvae vibriosis micro-encapsulated diet described in embodiment 1 or 2 or 3: anti-prawn larvae vibriosis Yolk antibody method for preparing freeze-dried powder step 4. in, during freeze drying, egg yolk liquid thickness 6mm, freeze-drying time is 10h.
Embodiment 5, in the anti-prawn larvae vibriosis micro-encapsulated diet described in embodiment 1 or 2 or 3 or 4: the particle diameter of described astragalus membranaceus powder is at 800 ~ 1000 orders, and the purity of described beta-schardinger dextrin-is not less than 80%.
Embodiment 6, in the anti-prawn larvae vibriosis micro-encapsulated diet of embodiment 1-5 described in any one: the mode adopting spraying dry (nozzle temperature is at 140 DEG C), obtained microcapsules dry powder feeding.
Embodiment 7, the application experiment of the present invention's anti-prawn larvae vibriosis micro-encapsulated diet and result thereof.
Experimental period: 2014 April ~ Mays in year.
Experiment place: Huaihai Institute of Technology aquatic animal disease laboratory, Jiangsu Province.
Animal used as test: Chinese prawn zoea and mysis.
(1) anti-prawn larvae vibriosis micro-encapsulated diet is used.During use with obtained antibody titer 2
10anti-prawn larvae vibriosis Yolk antibody freeze-dried powder 82.5%, particle diameter is at 900 object astragalus membranaceus powders 11%, and the beta-schardinger dextrin-6.5% of content more than 80%, above-mentioned raw materials spray-dried (nozzle temperature is at 140 DEG C) obtains microcapsules dry powder.The method for making of anti-prawn larvae vibriosis Yolk antibody freeze-dried powder is with reference to embodiment 1.
(2) experimental group Chinese prawn zoea and mysis (each 500 tails) the present invention that throws something and feeds anti-prawn larvae vibriosis microcapsule feed additive (30%) and commercially available prawn slice (70%), control group Chinese prawn zoea and mysis (each 500 tails) commercially available prawn slice of throwing something and feeding.
(3) throw something and feed after 4d, respectively determination experiment group and the Chinese prawn zoea of control group and the survival rate of mysis.
(4) throw something and feed after 4d, carry out protection inspection to experimental group and control group Chinese prawn zoea and mysis, get the young 150 tail respectively, carry out immersion infection with vibrio parahaemolytious and vibrio harveyi mixed bacteria liquid, thalline final concentration all reaches 1.8 × 10
6cFU/mL.Duration of test continues to throw something and feed by above-mentioned (2), and experimental period is 72h.
(5) experimental result: experimental group Chinese prawn zoea and mysis are thrown something and fed after anti-prawn larvae vibriosis micro-encapsulated diet (30%) described in experimental example 2 and commercially available prawn slice (70%), and can obtain higher resistance against diseases, survival rate is higher than control group by 34%.The experiment young vibrio parahaemolytious and vibrio harveyi mixed bacteria liquid carry out immersion infection, to throw something and feed the experimental group of anti-prawn larvae vibriosis microcapsule feed additive (30%) described in experimental example 2 and commercially available prawn slice (70%), survival rate 76% after young infection 72h, commercially available prawn slice control group larvae survival rate of throwing something and feeding is only 32%.Experimental result shows that anti-prawn larvae vibriosis micro-encapsulated diet can make Chinese prawn zoea and mysis obtain higher immune protective efficiency.
Claims (5)
1. an anti-prawn larvae vibriosis micro-encapsulated diet, is characterized in that, it forms micro-encapsulated diet by three kinds of raw materials of following weight proportion,
(1) anti-prawn larvae vibriosis Yolk antibody freeze-dried powder 80 ~ 85%;
(2) astragalus membranaceus powder 10% ~ 12%;
(3) beta-schardinger dextrin-5 ~ 8%;
Described anti-prawn larvae vibriosis Yolk antibody method for preparing freeze-dried powder step is as follows:
1. by cause prawn larvae vibriosis cause of disease vibrio parahaemolytious (
vibrioparahaemolyticus) VP1 and vibrio harveyi (
vibrioharveyi) kx1 bacterial strain is seeded on ordinary nutritional broth bouillon respectively, 24h cultivated by 28-30 DEG C of shaking table, adds formaldehyde 28 DEG C of deactivation 48h of volume final concentration 0.3%, centrifugal deactivation thalline;
2. decoct after getting Radix Astragali distilled water immersion 30-60min, filter, being concentrated into final concentration is former medicine 1g/mL, obtains Radix Astragali leachate;
3. mixed with Radix Astragali leachate by two kinds of deactivation thalline, obtained water aqua type vaccine, in vaccine liquid, cause of disease vibrio parahaemolytious VP1 and vibrio harveyi kx1 immunogene are respectively 10
11cells/mL, Radix Astragali leachate is 1g/mL;
4. with two kinds of obtained pathogenic vibrio water aqua type inactivated vaccines, adopt the adult laying hen of method immunity that is oral and collunarium combined immunization, the 1st time immunizing dose is: oral immunity joins in drinking-water by vaccine liquid, and addition is 5mL; Nasal immunization is that vaccine drop is entered laying hen nasal cavity, and infusion volume is 1mL; Press preceding method booster immunization, continuous booster immunization 5 times after one week, altogether immunity 6 times, immunity terminates latter 6th week, and Yolk antibody is tired and reached 2
10or more; To collect produce by immune chicken egg, isolate yolk, through freeze drying, obtained anti-prawn larvae vibriosis Yolk antibody freeze-dried powder.
2. anti-prawn larvae vibriosis micro-encapsulated diet according to claim 1, is characterized in that: anti-prawn larvae vibriosis Yolk antibody method for preparing freeze-dried powder step 4. in, during freeze drying, egg yolk liquid thickness 6mm, freeze-drying time is 10h.
3. anti-prawn larvae vibriosis micro-encapsulated diet according to claim 1, is characterized in that: the particle diameter of described astragalus membranaceus powder is at 800 ~ 1000 orders, and the purity of described beta-schardinger dextrin-is not less than 80%.
4. anti-prawn larvae vibriosis micro-encapsulated diet according to claim 1, is characterized in that: adopt spray-dired mode, obtained microcapsules dry powder feeding.
5. anti-prawn larvae vibriosis micro-encapsulated diet according to claim 1, it is characterized in that, the weight proportion of each raw material is as follows:
(1) anti-prawn larvae vibriosis Yolk antibody freeze-dried powder 82.5%;
(2) astragalus membranaceus powder 11%;
(3) beta-schardinger dextrin-6.5%.
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Cited By (3)
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| CN106924449A (en) * | 2017-03-30 | 2017-07-07 | 广州市普精生物科技有限公司 | One kind prevents and treats vibriosis Chinese medicine composition and its preparation method and application |
| CN109198267A (en) * | 2018-09-29 | 2019-01-15 | 扬州大学 | The compound additive and its preparation method and purposes of anti-Macrobrachium rosenbergii larvae enterobacteria disease |
| CN113005054A (en) * | 2021-02-04 | 2021-06-22 | 江苏海洋大学 | Bacillus amyloliquefaciens SS-ZC-26 and preparation method and application thereof |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106924449A (en) * | 2017-03-30 | 2017-07-07 | 广州市普精生物科技有限公司 | One kind prevents and treats vibriosis Chinese medicine composition and its preparation method and application |
| CN109198267A (en) * | 2018-09-29 | 2019-01-15 | 扬州大学 | The compound additive and its preparation method and purposes of anti-Macrobrachium rosenbergii larvae enterobacteria disease |
| CN113005054A (en) * | 2021-02-04 | 2021-06-22 | 江苏海洋大学 | Bacillus amyloliquefaciens SS-ZC-26 and preparation method and application thereof |
| CN113005054B (en) * | 2021-02-04 | 2022-07-12 | 江苏海洋大学 | Bacillus amyloliquefaciens SS-ZC-26 and preparation method and application thereof |
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Application publication date: 20151223 |