CN108094686A - The compound formulation and preparation method of aquatic livestock disease resistance can be effectively improved - Google Patents

The compound formulation and preparation method of aquatic livestock disease resistance can be effectively improved Download PDF

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Publication number
CN108094686A
CN108094686A CN201711386511.1A CN201711386511A CN108094686A CN 108094686 A CN108094686 A CN 108094686A CN 201711386511 A CN201711386511 A CN 201711386511A CN 108094686 A CN108094686 A CN 108094686A
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culture
powder
clostridium butyricum
bacterium
fermentation
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江国托
刘艳
单春乔
刘恩
曹艳子
张英雪
赵荣
高鑫
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DALIAN SANYI ANIMAL MEDICINE Co Ltd
JIANGSU SANYI BIOLOGICAL ENGINEERING CO LTD
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DALIAN SANYI ANIMAL MEDICINE Co Ltd
JIANGSU SANYI BIOLOGICAL ENGINEERING CO LTD
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
    • A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/111—Aromatic compounds
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/163—Sugars; Polysaccharides
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/174—Vitamins
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K50/00—Feeding-stuffs specially adapted for particular animals
    • A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
    • 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

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  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Polymers & Plastics (AREA)
  • Engineering & Computer Science (AREA)
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  • Food Science & Technology (AREA)
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  • Insects & Arthropods (AREA)
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Abstract

The invention discloses a kind of compound formulations and preparation method for effectively improving aquatic livestock disease resistance, and compound formulation each component and quality proportioning are as follows：Microbial germ powder 55 ~ 65, astragalus polyose 10 ~ 15, yeast cell wall 10 ~ 15, plants essential oil 5 ~ 10, vitamin B6 1 ~ 5；Each component mutual cooperation can reform intestinal microflora, change unrighted acid ratio in enteron aisle acid-producing bacteria metabolite, inhibit enteric pathogenic bacteria and other harmful substances, repair intestinal mucosa, nutrition digestion is promoted to absorb, body nospecific immunity can simultaneously enhanced, resisting stress enhances, there is stronger resistivity to adventitious viruses, germ, parasite and pollution environment, it is comprehensive to improve body bait utilization and survival rate, reduce residual bait, purify excrement, optimization and the intensive aquaculture model of specification.

Description

The compound formulation and preparation method of aquatic livestock disease resistance can be effectively improved
Technical field
The invention belongs to Applied Biotechnology fields, and in particular to a kind of to effectively improve answering for aquatic livestock disease resistance Close preparation and preparation method.
Background technology
With intensive culture pattern deepening continuously in industry, high-density breeding technology is increasingly subject to aquatic products field Concern.But due to non-standard operations such as breeding density is excessively high, bait feeding is excessive, abuse fishing medicines so that in water body and bed mud In the harmful substances such as excrement, residual bait, residual medicine constantly accumulate, various virosis, bacterial disease, parasitic disease in breeding environment not It is disconnected to grow.In order to prevent and treat various diseases in water body, many raisers select to launch antibiotic medicine, seriously The microecosystem balance of natural water and animal intestinal tract is destroyed, makes water body self-purification ability and cultivated animals disease resistance all big Amplitude reduction.At present, in order to preferably improve animal disease resistant ability, antibiotic usage is reduced, employs exempt from additional to organism Epidemic disease reinforcing agent improves the means of biological nospecific immunity, but the immunopotentiator from outside addition is not easy to disappear in enteron aisle Change and absorb, therefore the disease resistance raising of aquatic livestock is limited.
The content of the invention
For the present invention in order to solve the above-mentioned technical problem present in the prior art, aquatic livestock can be effectively improved by providing one kind The compound formulation and preparation method of disease resistance.
The present invention technical solution be：A kind of compound formulation for effectively improving aquatic livestock disease resistance, it is special Sign is that each component and quality proportioning are as follows：Microbial germ powder 55 ~ 65, astragalus polyose 10 ~ 15, yeast cell wall 10 ~ 15, plant Essential oil 5 ~ 10, vitamin B6 1 ~ 5；The microbial germ powder is by mortierella Diding bacterium powder（Acremonium terricola）With Clostridium butyricum powder（Clostridium butyricum）In mass ratio 35 ~ 50：15 ~ 30 mix, and the yeast cell wall is Bread leaven matricyte wall containing 30% beta glucan, the plants essential oil press quality by cinnaldehydrum, Thymol, lavender essential oil Than 2 ~ 4：2~4：1 ~ 2 mixes；Mortierella Diding bacterium powder active ingredient >=200mg/g, clostridium butyricum powder in the compound formulation Viable count >=5 × 108CFU/g, the mortierella Diding bacterium deposit number ATCC13215（NBRC30538）And it is protected through Chinese strain Administrative center CICC identifications are hidden, the clostridium butyricum deposit number is CICCNo.M 23847.
A kind of preparation method of the above-mentioned compound formulation for effectively improving aquatic livestock disease resistance, it is characterised in that specific Step is as follows：
A prepares microbial germ powder
（1）Prepare mortierella Diding bacterium powder
A. actication of culture
Mortierella Diding bacterium freeze-dried powder is seeded in PDA culture medium, 25 DEG C are cultivated 7 days；
B. seed culture
Strain after activation is transferred with 2% inoculum concentration in shaking flask, 25 DEG C of 24 ~ 36h of constant-temperature table shaken cultivation obtain seed Culture solution, the mass percent of fluid nutrient medium component are：White granulated sugar 1.4%, dried silkworm chrysalis meal 2.3%, dusty yeast 1.2%, more than water Amount, pH value 6.5；
C. fermentation tank culture
Seed culture fluid accesses 50L fermentation tanks with 20% inoculum concentration, and loading amount coefficient is 70%, ventilates and cultivates through 48 ~ 60h Tank is put, the mass percent of Fermenter Medium Component is：Analysis for soybean powder 2.8%, white granulated sugar 2.5%, dusty yeast 1.3%, dried silkworm chrysalis meal 2.5%、MgSO4 0.05%、KH2PO40.05%th, antifoaming agent 0.05%, water surplus are 6.5 with saturation NaOH tune pH value；
D. solid fermentation culture
The zymocyte liquid that step c is obtained is with 20%（v/m）Inoculum concentration access sterilizing after solid fermentation culture medium on, solid The constituent mass percentage of culture medium is：Corn flour 55%, dregs of beans 32%, wheat bran 13%；Condition of culture is：25 DEG C, humidity 70%, Fermentation time 72h；
E. the collection of mortierella Diding culture
After treating solid fermentation, culture is collected, direct 60 ~ 80 DEG C are dried to moisture ＜ 10%, are then crushed and mistake The mortierella Diding bacterium powder of the active ingredients such as 40 mesh sieves, as 94mg/g containing polysaccharide, polypeptide 28mg/g and biomass 35mg/g.
（2）Prepare clostridium butyricum bacterium powder
A actication of culture
Picking clostridium butyricum freeze-dried powder accesses activation medium, stands Anaerobic culturel；Culture medium forms：Peptone 10g, beef leaching Cream 10g, yeast extract 3g, glucose 10g, NaCl 5g, NaAc 3g, cysteine hydrochloride 0.5g, CaCO33g, distilled water are fixed Hold to 1000mL, 7.0,37 DEG C of cultures of pH for 24 hours, are cultivated with 2% inoculum concentration switching under new activation medium, the same terms 48h；
B. seed culture
Bacterium solution is accessed into 10L seeding tanks with 10% inoculum concentration, loading amount coefficient is 70%, and seed culture medium composition is：Peptone 5 ~ 10g, 5 ~ 10g of beef extract, 3 ~ 10g of yeast extract, glucose 5 ~ 10g, NaCl 5g, NaAc1 ~ 3g, CaCO3 1 ~ 3g, distillation Water is settled to 1000mL, and 7.0,37 DEG C of pH stands Anaerobic culturel 20h；
C. fermentation tank culture
Seed culture fluid accesses 100L fermentation tanks with 10% inoculum concentration, and loading amount coefficient is 70%, and fermentation tank culture medium composition is： 5 ~ 10g of peptone, 3 ~ 10g of yeast extract, 5 ~ 15g of glucose, starch 0.5 ~ 1g, NaCl 5g, NaAc 1 ~ 3g, CaCO3 3~ 10g, water are settled to 1000mL, and 7.0,37 DEG C of pH stands Anaerobic culturel for 24 hours；
D. microorganism collection
Zymotic fluid is collected, supernatant is abandoned after centrifugation, obtains wet thallus；By clostridium butyricum wet thallus and dried starch with mass ratio 1: 0.5 ~ 5 mixing, for 24 hours, pulverizer crushes for 50 DEG C of dryings, crosses 60 mesh sieves, adjusts viable count >=1 × 10 of clostridium butyricum10CFU/g, as Clostridium butyricum opportunistic pathogen powder；
B is respectively by astragalus polyose, yeast cell wall, vitamin B660 mesh sieves are smashed it through, it is afterwards that astragalus polyose, yeast is thin Cell wall, vitamin B6, mortierella Diding culture, clostridium butyricum powder and plants essential oil be put into mixing plant, stir evenly.
The present invention is by mortierella Diding, clostridium butyricum, astragalus polyose, yeast cell wall, plants essential oil and vitamin B6It is mixed It closes, each component mutual cooperation can reform intestinal microflora, change unsaturated fat in enteron aisle acid-producing bacteria metabolite Sour ratio inhibits enteric pathogenic bacteria and other harmful substances, repairs intestinal mucosa, and nutrition digestion is promoted to absorb, while can make machine Body nospecific immunity enhances, resisting stress enhancing, has stronger resistance to adventitious viruses, germ, parasite and pollution environment Ability, it is comprehensive to improve body bait utilization and survival rate, residual bait is reduced, purifies excrement, optimization and specification intensive culture mould Formula.
Specific embodiment
Embodiment 1：
The compound formulation for effectively improving aquatic livestock disease resistance of the present invention, each component and quality proportioning are as follows：Microorganism Bacterium powder 65, astragalus polyose 15, yeast cell wall 15, plants essential oil 10, vitamin 5；The microbial germ powder is by mortierella Diding Bacterium powder（Acremonium terricola）With clostridium butyricum powder（Clostridium butyricum）In mass ratio 50：30 is mixed It closes, the yeast cell wall is the bread leaven matricyte wall containing 30% beta glucan, and the plants essential oil is by cinnaldehydrum, hundred In fragrant phenol, lavender essential oil in mass ratio 2：2：1 mixes；Mortierella Diding bacterium powder active ingredient in the compound formulation >= 200mg/g, clostridium butyricum powder viable count >=5 × 108CFU/g, the mortierella Diding bacterium deposit number ATCC13215 （NBRC30538）, and identified through Chinese culture presevation administrative center CICC, the clostridium butyricum deposit number is CICCNo.M 23847。
A kind of preparation method of the above-mentioned compound formulation for effectively improving aquatic livestock disease resistance, it is characterised in that specific Step is as follows：
A prepares microbial germ powder
（1）Prepare mortierella Diding bacterium powder
A. actication of culture
By mortierella Diding bacterium freeze-dried powder（Outsourcing）It is seeded in PDA culture medium, 25 DEG C are cultivated 7 days, i.e., a large amount of coremiums to be generated After transfer；
B. seed culture
Strain after activation is transferred with 2% inoculum concentration in shaking flask, 25 DEG C of 24 ~ 36h of constant-temperature table shaken cultivation obtain seed Culture solution, the mass percent of fluid nutrient medium component are：White granulated sugar 1.4%, dried silkworm chrysalis meal 2.3%, dusty yeast 1.2%, more than water Amount, pH value 6.5；
C. fermentation tank culture
Seed culture fluid accesses 50L fermentation tanks with 20% inoculum concentration, and loading amount coefficient is 70%, ventilates and cultivates through 48 ~ 60h, i.e., Mycelia raised growth and start produce spore can put tank, the mass percent of Fermenter Medium Component is：Analysis for soybean powder 2.8%, white sand Sugar 2.5%, dusty yeast 1.3%, dried silkworm chrysalis meal 2.5%, MgSO4 0.05%、KH2PO40.05%th, antifoaming agent 0.05%, water surplus, with full It is 6.5 with NaOH tune pH value；
D. solid fermentation culture
The zymocyte liquid that step c is obtained is with 20%（v/m）Inoculum concentration access sterilizing after solid fermentation culture medium on, solid The constituent mass percentage of culture medium is：Corn flour 55%, dregs of beans 32%, wheat bran 13%；Condition of culture is：25 DEG C, humidity 70%, Fermentation time 72h；
E. the collection of mortierella Diding culture
After treating solid fermentation, culture is collected, direct 60 ~ 80 DEG C are dried to moisture ＜ 10%, are then crushed and mistake The mortierella Diding bacterium powder of the active ingredients such as 40 mesh sieves, as 94mg/g containing polysaccharide, polypeptide 28mg/g and biomass 35mg/g.
（2）Prepare clostridium butyricum bacterium powder
A actication of culture
Picking clostridium butyricum freeze-dried powder（Outsourcing）Activation medium is accessed, stands Anaerobic culturel；Culture medium forms：Peptone 10g, Beef extract 10g, yeast extract 3g, glucose 10g, NaCl 5g, NaAc 3g, cysteine hydrochloride 0.5g, CaCO33g steams Distilled water is settled to 1000mL, and 7.0,37 DEG C of cultures of pH for 24 hours, are transferred with 2% inoculum concentration in new activation medium, the same terms Lower culture 48h, microscopy, more than 90% thalline form gemma；
B. seed culture
Bacterium solution is accessed into 10L seeding tanks with 10% inoculum concentration, loading amount coefficient is 70%, and seed culture medium composition is：Peptone 5 ~ 10g, 5 ~ 10g of beef extract, 3 ~ 10g of yeast extract, glucose 5 ~ 10g, NaCl 5g, NaAc1 ~ 3g, CaCO3 1 ~ 3g, distillation Water is settled to 1000mL, and 7.0,37 DEG C of pH stands Anaerobic culturel 20h；
C. fermentation tank culture
Seed culture fluid accesses 100L fermentation tanks with 10% inoculum concentration, and loading amount coefficient is 70%, and fermentation tank culture medium composition is： 5 ~ 10g of peptone, 3 ~ 10g of yeast extract, 5 ~ 15g of glucose, starch 0.5 ~ 1g, NaCl 5g, NaAc 1 ~ 3g, CaCO3 3~ 10g, water are settled to 1000mL, and 7.0,37 DEG C of pH stands Anaerobic culturel for 24 hours, and microscopy, more than 90% thalline forms gemma, that is, puts Tank terminates to cultivate；
D. microorganism collection
Zymotic fluid is collected, supernatant is abandoned after centrifugation, obtains wet thallus；
Clostridium butyricum wet thallus is mixed with dried starch with mass ratio 1: 0.5 ~ 5, for 24 hours, pulverizer crushes for 50 DEG C of dryings, crosses 60 mesh Sieve adjusts viable count >=1 × 10 of clostridium butyricum10CFU/g is clostridium butyricum opportunistic pathogen powder；
B is respectively by astragalus polyose, yeast cell wall, vitamin B660 mesh sieves are smashed it through, it is afterwards that astragalus polyose, yeast is thin Cell wall, vitamin B6, mortierella Diding culture, clostridium butyricum powder and plants essential oil be put into mixing plant, stir evenly.
Embodiment 2：
The compound formulation for effectively improving aquatic livestock disease resistance of the present invention, it is characterised in that each component and quality proportioning are such as Under：Microbial germ powder 55, astragalus polyose 10, yeast cell wall 10, plants essential oil 5, vitamin B6 1；The microbial germ powder is By mortierella Diding bacterium powder（Acremonium terricola）With clostridium butyricum powder（Clostridium butyricum）By quality Than 35：15 mix, the yeast cell wall be the bread leaven matricyte wall containing 30% beta glucan, the plants essential oil by Cinnaldehydrum, Thymol, lavender essential oil in mass ratio 2：2：1 mixes；There is mortierella Diding bacterium powder in the compound formulation Imitate ingredient >=200mg/g, clostridium butyricum powder viable count >=5 × 108CFU/g, the mortierella Diding bacterium deposit number ATCC13215 （NBRC30538）, and identified through Chinese culture presevation administrative center CICC, the clostridium butyricum deposit number is CICCNo.M 23847。
Preparation method is the same as embodiment 1.
Embodiment 3：
The compound formulation for effectively improving aquatic livestock disease resistance of the present invention, it is characterised in that each component and quality proportioning are such as Under：Microbial germ powder 60, astragalus polyose 12, yeast cell wall 13, plants essential oil 8, vitamin B6 3；The microbial germ powder is By mortierella Diding bacterium powder（Acremonium terricola）With clostridium butyricum powder（Clostridium butyricum）By quality Than 40：20 mix, the yeast cell wall be the bread leaven matricyte wall containing 30% beta glucan, the plants essential oil by Cinnaldehydrum, Thymol, lavender essential oil in mass ratio 3：3：2 mix；There is mortierella Diding bacterium powder in the compound formulation Imitate ingredient >=200mg/g, clostridium butyricum powder viable count >=5 × 108CFU/g, the mortierella Diding bacterium deposit number ATCC13215 （NBRC30538）, and identified through Chinese culture presevation administrative center CICC, the clostridium butyricum deposit number is CICCNo.M 23847。
Preparation method is the same as embodiment 1.
The compound formulation of the present invention is used for feeding additive aquatic animal, and application method is as follows：Additive amount is feed gross mass 0.05%~0.3%, it can be directly appended to be mixed and made into pre-mixing agent in aquatic animal feed or with carrier；Or add with other feeds Agent or feedstuff is added to be mixed and made into premix, concentrate feed form feeding aquatic livestock.
Experimental example one：
Carp similar in selecting specification, is randomly divided into four groups：Complex group, bacterium powder group, immunizing agent group and blank control group, in interior It is fed at 20 ~ 25 DEG C of water temperature in aquarium, every group 100, feeding cycle is 100 days.Each group feeds same underlying commodity feed. Complex group adds the compound formulation of 0.06% embodiment of the present invention, and bacterium powder group is added micro- in 0.06% embodiment of the present invention 1 Biological bacteria powder, immunizing agent group addition Radix Astragali 0.025%, yeast cell wall 0.025%, plants essential oil 0.01%, blank control group Only addition underlying commodity feed, other rearing conditions are identical.Fish growing state and water quality condition are recorded, is shown in Table 1.
Table 1
Group Weight gain rate（%） Fish body survival rate（%） Feed coefficient Fish body vigor Water colour
Complex group 80 99 2.21 It is active Clearly
Bacterium powder group 70 95 2.30 It is more active Clearly
Immunopotentiator 65 94 2.52 It is active It is muddy
Blank control group 60 90 2.55 It is dull It is muddy
Result of the test show to add the complex group weight gain of compound formulation of the embodiment of the present invention, survival rate etc. apparently higher than its His group（Bacterium powder group, immunologic facilitation group and blank control group）, feed coefficient is substantially less than control group.Illustrate feed addition of the present invention Agent can promote carp to digest, beneficial to feed, so as to which body be promoted to increase.Compared with the control group, experimental group water quality is preferable, and water colour is steady It is fixed,
Experimental example two：
Pond crucian carp fish similar in selecting specification, is randomly divided into four groups:Complex group, bacterium powder group, immunizing agent group and blank control group, every group 200, cultivating condition is the same as experimental example one.Each group adds same underlying commodity feed, and 0.1% present invention of complex group addition is implemented The compound formulation of example 1, bacterium powder group add the microbial germ powder in 0.1% embodiment of the present invention 1, immunizing agent group addition Radix Astragali 0.045%th, yeast cell wall 0.045%, plants essential oil 0.01%, blank control group only add underlying commodity feed, other feedings The condition of supporting is identical.Record fish growing state, water quality condition and quality to pond crucian carp fish etc. do further detection and analysis, the results are shown in Table 2。
Table 2
Result of the test shows：Compared with the control group, complex group pond crucian carp daily gain and survival rate propose different degrees of raising, in meat Crude fat content is remarkably decreased, and superoxide dismutase activity improves a lot compared with control group in serum.Thus illustrate, the present invention Compound formulation can effectively facilitate pond crucian carp ingest and digestion power, increase survival rate, and immunity can be significantly improved.
Experimental example three：
By 400 uniform in size, the comparable rainbow trout of weight is put at random in 4 mouthfuls of culturing jars, per 100, cylinder, 4 mouthfuls of cylinders are randomly divided into Four groups:Complex group, bacterium powder group, immunizing agent group and blank control group.Complex group adds 0.1% embodiment of the present invention, 2 compound system Agent, bacterium powder group, immunizing agent group and blank control group are the same as experimental example two.Record food ration, water temperature are controlled at 16 ~ 18 DEG C in detail, are protected Hold continuous oxygenation.It after cultivation in 35 days, when fasting 24 is small, weighs one by one, amount body is long, the results are shown in Table 3.
Table 3
Group Weight gain rate（%） Fish body survival rate（%） Feed coefficient Fish body vigor Water colour
Complex group 85 100 1.87 It is active Clearly
Bacterium powder group 72 97 1.89 It is more active Clearly
Immunopotentiator 66 95 2.11 It is active It is muddy
Control group 59 89 2.32 It is dull It is muddy
Result of the test shows：In complex group the weightening of rainbow trout and survival rate it is different degrees of higher than control group, illustrate that invention is multiple The growth performance of rainbow trout can effectively be improved by closing preparation, and the feed coefficient of complex group is lower compared with control group, illustrates institute of the present invention The compound formulation of offer can preferably improve efficiency of feed utilization.
Experimental example four：
The compound formulation of the embodiment of the present invention 2 is added in penaeus vannamei boone feed according to 0.3% and 0.1% proportioning, is raised Support 100 days phases.Experimental result is shown:The complex group of addition 0.3% improves 18.9% than control composition motility rate, feed coefficient drop Low 0.16；It adds 0.1% feed addictive group and improves 13.2% than control composition motility rate, feed coefficient reduces 0.12. The experiment Analysis of variance, complex group reach the level of signifiance with control group difference（P ＜ 0.05）Experimental result is shown in Table 4.
Table 4
Group Weight gain（%） Shrimp body survival rate（%） Feed coefficient Shrimp vitality of subject Water colour
0.3% group 81 99.9 2.01 It is active Clearly
0.1% group 76 94.2 2.15 It is more active Clearly
Control group 68 81.0 2.17 It is dull It is muddy
Experimental example five
Four ponds of certain farm are chosen, the sizable rice fish of weight is raised, is divided into four groups:Complex group, bacterium powder group are exempted from Epidemic disease agent group and blank control group.Complex group adds the compound formulation of 0.1% embodiment of the present invention 3, bacterium powder group, immunizing agent group and sky White control group is the same as embodiment two.Four groups of cultivating conditions are identical, and the test period is 100 days, counts the survival of rice fish in every group Rate, average weight simultaneously record water quality situation, while quality to rice fish etc. further tests and analyzes, and the results are shown in Table 5.
Table 5
Group Weight gain rate（%） Fish body survival rate（%） Fat content Fish body vigor Water colour
Complex group 87 100 14.34 It is active Clearly
Bacterium powder group 71 95 14.44 It is more active Clearly
Immunopotentiator group 66 90 15.56 It is active It is muddy
Control group 51 69 15.79 It is dull It is muddy
Result of the test is shown：Obviously higher than control group, fat content is significantly lower than for complex group rice fish survival rate and average weight Control group, there is not the situation of fry death in the breeding process in complex group, while microbial inoculum is stronger for bottom of pond clean-up capability, Quantity of exchanged water is reduced, water quality is more stable.And compare pool cultivated advanced fry and gastro-intestinal problems occur, there is mortality in fry.From And illustrate that compound formulation of the present invention is conducive to a meter fish growth, can effectively improve survival rate, and significantly improve disease resistance.
Experimental example six：
Clinical trial is done in certain farm, the compound formulation of the addition embodiment of the present invention 3 is used to treat the eel arc of rainbow trout infection Bacterium disease, sick fish performance：Fin and the hyperemia of gill portion are rubescent, and muscle groups are woven with Mass or petechial hemorrhage, and dissection has yellow when examining Sticky ascites, intestinal submucosa tissue damage, part fish hepatic necrosis.The fishpond death rate is 5%.By the compound of the embodiment of the present invention 3 System is added to 0.3% additive amount in fish basal feed, is fed daily 3 times, is used continuously 30 days, treated effect reaches 70 ~ 85%, do not occur dead fish phenomenon.

Claims (2)

1. a kind of compound formulation for effectively improving aquatic livestock disease resistance, it is characterised in that each component and quality proportioning are such as Under：Microbial germ powder 55 ~ 65, astragalus polyose 10 ~ 15, yeast cell wall 10 ~ 15, plants essential oil 5 ~ 10, vitamin B6 1 ~ 5；Institute It is by mortierella Diding bacterium powder to state microbial germ powder（Acremonium terricola）With clostridium butyricum powder（Clostridium butyricum）In mass ratio 35 ~ 50：15 ~ 30 mix, and the yeast cell wall is the bread ferment containing 30% beta glucan Mother cell wall, the plants essential oil is by cinnaldehydrum, Thymol, lavender essential oil in mass ratio 2 ~ 4：2~4：1 ~ 2 mixes； Mortierella Diding bacterium powder active ingredient >=200mg/g in the compound formulation, clostridium butyricum powder viable count >=5 × 108CFU/g, institute State mortierella Diding bacterium deposit number ATCC13215（NBRC30538）And it is identified through Chinese culture presevation administrative center CICC, institute Clostridium butyricum deposit number is stated as CICC No.M 23847.
2. a kind of preparation method for the compound formulation that can effectively improve aquatic livestock disease resistance as described in claim 1, special Sign is to be as follows：
A prepares microbial germ powder
（1）Prepare mortierella Diding bacterium powder
A. actication of culture
Mortierella Diding bacterium freeze-dried powder is seeded in PDA culture medium, 25 DEG C are cultivated 7 days；
B. seed culture
Strain after activation is transferred with 2% inoculum concentration in shaking flask, 25 DEG C of 24 ~ 36h of constant-temperature table shaken cultivation obtain seed Culture solution, the mass percent of fluid nutrient medium component are：White granulated sugar 1.4%, dried silkworm chrysalis meal 2.3%, dusty yeast 1.2%, more than water Amount, pH value 6.5；
C. fermentation tank culture
Seed culture fluid accesses 50L fermentation tanks with 20% inoculum concentration, and loading amount coefficient is 70%, ventilates and cultivates through 48 ~ 60h Tank is put, the mass percent of Fermenter Medium Component is：Analysis for soybean powder 2.8%, white granulated sugar 2.5%, dusty yeast 1.3%, dried silkworm chrysalis meal 2.5%、MgSO4 0.05%、KH2PO40.05%th, antifoaming agent 0.05%, water surplus are 6.5 with saturation NaOH tune pH value；
D. solid fermentation culture
The zymocyte liquid that step c is obtained is with 20%（v/m）Inoculum concentration access sterilizing after solid fermentation culture medium on, solid The constituent mass percentage of culture medium is：Corn flour 55%, dregs of beans 32%, wheat bran 13%；Condition of culture is：25 DEG C, humidity 70%, Fermentation time 72h；
E. the collection of mortierella Diding culture
After treating solid fermentation, culture is collected, direct 60 ~ 80 DEG C are dried to moisture ＜ 10%, are then crushed and mistake The mortierella Diding bacterium powder of the active ingredients such as 40 mesh sieves, as 94mg/g containing polysaccharide, polypeptide 28mg/g and biomass 35mg/g；
（2）Prepare clostridium butyricum bacterium powder
A actication of culture
Picking clostridium butyricum freeze-dried powder accesses activation medium, stands Anaerobic culturel；Culture medium forms：Peptone 10g, beef leaching Cream 10g, yeast extract 3g, glucose 10g, NaCl 5g, NaAc 3g, cysteine hydrochloride 0.5g, CaCO33g, distilled water are fixed Hold to 1000mL, 7.0,37 DEG C of cultures of pH for 24 hours, are cultivated with 2% inoculum concentration switching under new activation medium, the same terms 48h；
B. seed culture
Bacterium solution is accessed into 10L seeding tanks with 10% inoculum concentration, loading amount coefficient is 70%, and seed culture medium composition is：Peptone 5 ~ 10g, 5 ~ 10g of beef extract, 3 ~ 10g of yeast extract, glucose 5 ~ 10g, NaCl 5g, NaAc1 ~ 3g, CaCO3 1 ~ 3g, distillation Water is settled to 1000mL, and 7.0,37 DEG C of pH stands Anaerobic culturel 20h；
C. fermentation tank culture
Seed culture fluid accesses 100L fermentation tanks with 10% inoculum concentration, and loading amount coefficient is 70%, and fermentation tank culture medium composition is： 5 ~ 10g of peptone, 3 ~ 10g of yeast extract, 5 ~ 15g of glucose, starch 0.5 ~ 1g, NaCl 5g, NaAc 1 ~ 3g, CaCO3 3~ 10g, water are settled to 1000mL, and 7.0,37 DEG C of pH stands Anaerobic culturel for 24 hours；
D. microorganism collection
Zymotic fluid is collected, supernatant is abandoned after centrifugation, obtains wet thallus；By clostridium butyricum wet thallus and dried starch with mass ratio 1: 0.5 ~ 5 mixing, for 24 hours, pulverizer crushes for 50 DEG C of dryings, crosses 60 mesh sieves, adjusts viable count >=1 × 10 of clostridium butyricum10CFU/g, as Clostridium butyricum opportunistic pathogen powder；
B is respectively by astragalus polyose, yeast cell wall, vitamin B660 mesh sieves are smashed it through, afterwards by astragalus polyose, yeast cells Wall, vitamin B6, mortierella Diding culture, clostridium butyricum powder and plants essential oil be put into mixing plant, stir evenly.
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