CN109055266A - A kind of biological agent and the preparation method and application thereof with vibrios growth in the inhibition shrimp pool - Google Patents
A kind of biological agent and the preparation method and application thereof with vibrios growth in the inhibition shrimp pool Download PDFInfo
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- CN109055266A CN109055266A CN201810950322.0A CN201810950322A CN109055266A CN 109055266 A CN109055266 A CN 109055266A CN 201810950322 A CN201810950322 A CN 201810950322A CN 109055266 A CN109055266 A CN 109055266A
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Abstract
The present invention provides it is a kind of with inhibit the shrimp pool in vibrios growth biological agent leavening, the leavening include bacterial content account for the bacteriophagic Bdellovibrio of total bacteria count 5%~15%, 5%~15% Bdellovibrio starrii, 20%~40% bacillus, 20%~40% lactic acid bacteria and 10%~30% ocean rhodotorula.The present invention be also extracted it is a kind of with inhibit the biological agent of vibrios growth in the shrimp pool with and preparation method thereof, the biological agent is mainly by the activated post-fermentation raw material preparation gained of above-mentioned leavening.The present invention is prepared into biological agent by reasonably combined a variety of probiotics fermentation raw materials, cooperate to obtain a kind of biological agent for effectively inhibiting vibrios growth in the shrimp pool using different strain, the preparation method of said preparation is simple, it is not high to equipment requirement, strong operability is promoted the use of easily in scale raising or in aquaculture cooperative society.
Description
Technical field
The present invention relates to aquaculture water quality regulation and control fields, and in particular to a kind of with the life for inhibiting vibrios growth in the shrimp pool
Object preparation and the preparation method and application thereof.
Background technique
Prawn culturing is mainstay industry important in China's culture fishery.But recently as the increasing of cultured output
Add, the increasing of cultivation density, the large-scale development of prawn culturing, aquaculture pollution aggravation causes breeding environment quality continuous
Deteriorate, causes causal organism type to increase and accelerate with spread speed, the farming disease harms are on the rise, and the disease in prawn culturing is special
It is that bacterial disease occurs frequently and influences seriously, to cause huge economic loss, significantly limit China's culture fishery
Development.The farming disease harms problem and breeding environment problem have become one of the bottleneck factor for restricting mariculture industry development, seriously
The development that annoying China's mariculture industry constrains the sound development of blue agriculture.
Vibriosis is a kind of phytosis that is most common, endangering most serious in sea-farming, mainly by vibrios (Vibrio)
Cause, returns in " Berger bacterial identification manual " of (and the whole world is most authoritative) division bacteria which generallys use
Belong to Prokaryota, Bacteriophyta, gram facultative Bacteroides nodosus mesh, vibrionaceae, (vibrio, Aeromonas, Plesiomonas
Belong to, Photobacterium, Lucibacterium).Named at present 91 kinds.Common kinds of pathogenic vibrio has Vibrio anguillarum (Vibrio
Anguillarum), Vibrio harveyi (V.harveyi), vibrio alginolyticus (V.alginolyticus), vibrio parahaemolytious
(V.parahaemolyticus) and Vibrio vulnificus (V.vulnificus) etc..It is dynamic that some researches show that nearly all sea-farmings
Object, such as prawn, swimming crab, grouper, red porgy, Larimichthys crocea, lefteye flounder, Zhang Xiongyu (seriola dumerili) suffer from vibriosis, can cause pair
Blind's disease of shrimp, kwashiorkor, fin rot, Tou dead sick (EMS) etc., can also cause complication and secondary disease of various diseases etc..
It if vibrio parahemolyticus is a kind of maritime bacterium, is widely distributed in seawater and mariculture products, China East China edge
In the seawater of bank vibrio parahemolyticus recall rate be 47.5%-66.5%, extra large water fish, shrimp average bacterial bearing rate be 45.6%-
48.7%, summer may be up to 90% or more.Hemotoxin has direct hemolytic, can make prawn various kinds of cell that haemolysis occur, molten
Solve prawn blood, cell-lethal.Thermostable direc t hemolysin can cause prawn digestive system tissue damage, and ileum is rotten to the corn, gastric mucosa
Inflammation or even liver pancreas, the disorders of digestion.Somebody thinks that vibrio parahemolyticus generates urase, generates the accumulation of prawn intestinal juice
Enteron aisle ponding, serious person, which is likely to occur, draws white just phenomenon.Under the extracellular products of vibrio parahemolyticus can make immunity of prawn obvious
Drop, the speed of growth is slow, so that easily causing other a variety of diseases.
The main means controlled at present aquiculture animal bacterial disease are largely resisted using antibacterial drug therapy
Raw element medicine class is applied to each stage of breeding process.It is not comprehensive due to recognizing aetology, pharmacology etc., it is cultivating
There are a variety of drugs of high dose in the process the abuse such as to be used in mixed way, abuse Phenomenon of Drugs, brings serious drug resistance, drug resistance, with
And antibiotic residue problem, cause social extensive concern.Based on current prawn culturing produces still in a manner of pond culture, including
Intertidal zone comprehensive ecological cultivation mode, half intensive culture mode of common shrimp ponds, spreading feed coefficient intensive culture mode etc..By more than ten years
Development, summarizes its culture efficiency, discovery still remain regulating and controlling water quality is not easy, eutrophication is serious, unit of water body cultured output not
High disadvantage, the especially influence also vulnerable to burst factors such as temperature season limit, storm tide invasion, development space is by larger system
About.Recently as the continuous promotion of cultivation scale, the uncontrolled utilization of water resource, pond culture mode faces water environment dirt
Dye, disease increase year by year, food safety demand, coastal industry land used squeeze and country advocates the pressure such as energy-saving and emission-reduction.
Since the technology of usual used physico-chemical method control water quality has various drawbacks, so biology is (mainly micro-
Biology) it controls and is just paid attention to.Micro ecological technology regulates and controls water quality, control cause of disease by beneficial microbe, and enhancing is immune, improves
Nutrition, at the same reduce or do not have to chemicals, have many advantages, such as it is at low cost, bring notable results, it is pollution-free, it has also become current prawn culturing
One of main developing direction of technology has generallyd use in prawn culturing and has obtained significant benefit.The use of microorganism, can
To improve cultured prawn immunity of organisms, substance cyclic metabolism can also be accelerated, purifying water body improves the health of breeding water body.
The principle of microbiological treatment breeding wastewater is: using the albumen, carbohydrate, fatty acid of heterotrophicy bacteria mineralising solubility at nothing
Machine salt, using nitrobacteria by NH4+、NO2It is converted into the NO of low toxicity3, recycle denitrifying bacteria by NO3Denitrogenation is to N2。
In breeding wastewater biochemical treatment at present using it is more be bio-contact oxidation (Liao Shaoan etc., 2008), biological rotating disk
The techniques such as (Brazil, 2006), biological fluidized bed (Bortone et al, 2003), the in these processes big multi-source of biofacies
The ligotrophic group in nature or artificial environment does not adapt to the nutrient environment of breeding wastewater, the unit of microbiological treatment module
Volume processing load is not practical high, is greatly affected the application of traditional microbiological Treatment process.
Water quality is generally controlled using single or complex microorganism bacterial classification both at home and abroad at present, but since microorganism is by extraneous ring
Border is affected, and anti-poor environment impact capacity is poor, once system damage is difficult to restore, therefore treatment effect is unstable.Therefore
It needs the type to microorganism and constitutes to carry out scientific matching, to improve its efficiency.Wu Wei etc. (2001) false unit cell red to marsh
Bacterium, 3 kinds of bacterial strains of Nocard's bacillus and Candida it is same surely change cell ammonia nitrogen in breeding water body and cultured water are converted, send out
If existing be combined into compound strain and immobilization by 2:1:2 for 3 bacterial strains, to the ammonia nitrogen and cultured water conversion effect in breeding water body
Fruit will more preferably.
Therefore this invention address that developing a kind of with the biological agent for inhibiting vibrios growth in the shrimp pool, biological agent master
It to include Bdellovibrio, bacillus, ocean rhodotorula, lactic acid bacteria.
Bdellovibrio belongs to Bdellovibrio section, is widely present in nature, soil, river water, neighbouring maritime waters and effluent sewerage
In be all distributed, 10/milliliter containing bacterium number in general sewage can contain ten thousand/milliliter of bacterium number in effluent sewerage, contain bacterium in soil
10/gram of number has all germy morphological features: unicellular, arc or comma shape, sometimes in the shape of a spiral.Size is 0.3-
0.6 × 0.8-1.2 microns, or the 1/3-1/4 of only bacillus length.Raw flagellum is held seldom to be more than one, some is raw in the other end
There is a branch of cilium.The flagellum of aquatic Bdellovibrio also has sheath, it is the extension of cell wall, and surrounds flagellum filamentous, institute
With thick 3-4 times of flagellum than other bacteriums, this is a significant feature.Bdellovibrio movement is active, Gram-negative.
Protein content is higher in cell, and some accounts for the 60-70% of dry weight.DNA content 5%, GC percentage are 42-51.It has been reported that leech
The synthesis of vibrios DNA is carried out after host cell DNA cracks completely, and 80% DNA is and into bacterial parasite in host cell.
The bacterium does not depend on extraneous source.Therefore, even if the place seldom in nutriment, the ecosystems such as ocean, rivers, lake
It can also be grown in system.
Bacillus (Bacillus) is different from vegetative cell, resistant strong spy in structure and chemical component
Point has very strong resistance, the marine extreme environments such as energy resistance to oxidation, anti-extrusion, high temperature resistant to high temperature, ultraviolet light, drying etc..Together
When bacillus can generate multiple active enzymes, have improve aquiculture animal growth performance characteristic.Sugita etc. has found bud
Spore bacillus can provide some nutriments and promote the development of lefteye flounder enteron aisle and the expression of digestive ferment, to promote the suction of nutriment
It receives.Xu Haiyan is research shows that bacillus subtilis generates the volatile fats such as acetic acid, propionic acid, butyric acid during its growth and breeding
Acid reduces intestinal pH, inhibits harmful bacteria growth, provides acid condition for beneficial bacterium growth, promotes growth of animal, improve productivity
Energy.The quick equal processing for just studying simple biofilter and microorganism formulation for the processing of Macrobrachium rosenbergii breeding water body in river, as a result sends out
The biofilter now cured can effectively improve the water quality of nursery water, mitigate the organic loading of water body, reduce ammonia nitrogen and nitrite nitrogen
Content has apparent growth promoting effect to Macrobrachium rosenbergii larvae, can reduce water body ammonia-nitrogen content to a certain extent, promotes Roche
The growth of the pond crayfish young.
Ocean rhodotorula (Rhodotorula benthica) is the single celled eukaryotic life separated from the bed mud of ocean
Object, the several physiological active substances such as protein, carbohydrate, vitamin, digestive ferment and growth factor rich in, be fish, shrimp,
The good open-mouthed bait of the economic animals such as shellfish, sea cucumber.Practice discovery, ocean rhodotorula can significantly improve depositing for aquatic livestock seedling
Motility rate, reduces antibiotic dosage at the immune function for improving feed efficiency and the price of deed and capable of enhancing animal body, is ecologic breeding
Excellent additive.The aquatic products for adding ocean rhodotorula feed of ingesting have that disease resistance is strong, survival rate is high, skin and muscle
The features such as lovely luster, features good taste.
Lactic acid bacteria (Lacticacid bacteria) is that a group form, metabolism performance and physiologic character are not exactly the same
Gram-positive bacteria general designation.Growth of animal and immunostimulation mechanism can be promoted to be divided into two kinds lactic acid bacteria at present: first is that straight
Connect effect, i.e. thallus itself and its active metabolite sticking or degrade to harmful substance；Second is that indirectly-acting, i.e. lactic acid bacteria
It adjusts the modes such as immunity of organism activity and body part enzymatic activity by adjusting intestinal flora and its metabolic enzyme activity and plays indirectly
Prebiotic effect.Lactic acid bacteria can generate B family vitamin, including folic acid, biotin, vitamin B and vitamin K etc., can directly be
Host provides required nutriment and promotes growth of animal.In addition, the acidic metabolite of lactic acid bacteria can reinforce intestines peristalsis, have
Conducive to the digestion and absorption of nutrient.The antibacterial materials such as organic acid, hydrogen peroxide can be generated during the growth metabolism of lactic acid bacteria, are reached
To the purpose for inhibiting pathogenic bacteria and the growth of exogenous pathogen bacterium.Lactic acid bacteria can prevent pathogen in the attachment of enterocyte surface, determine
Intestinal cell is planted and invades, this mechanism is referred to as " sticking resistance ".
Summary of the invention
In view of this, the present invention provide it is a kind of with inhibit in the shrimp pool biological agent and preparation method thereof of vibrios growth with
Using the biological agent is by utilizing bacteriophagic Bdellovibrio, Bdellovibrio starrii, bacillus, ocean rhodotorula and lactic acid bacteria
Compound formulation is made in scientific matching, and said preparation has good effect to vibrios growth in the inhibition shrimp pool, and easy to use, suppression
Vibrios effect processed is obvious.
To inhibit vibrios growth in the shrimp pool, first aspect present invention provides a kind of with vibrios growth in the inhibition shrimp pool
The leavening of biological agent, including compound bacteria A and compound bacteria B；
Wherein, the compound bacteria A include bacteriophagic Bdellovibrio, Bdellovibrio starrii it is one or more；The compound bacteria B packet
Include one of bacillus, lactic acid bacteria, ocean rhodotorula or a variety of.
In an embodiment of the present invention, when described a kind of with the leavening for inhibiting the biological agent of vibrios growth in the shrimp pool
When including bacteriophagic Bdellovibrio, Bdellovibrio starrii, bacillus, lactic acid bacteria, ocean rhodotorula, the bacteriophagic Bdellovibrio content is accounted for
The percentage of total bacteria count is 5%~15%；The percentage that the Bdellovibrio starrii content accounts for total bacteria count is 5%~15%；Institute
Stating bacillus content and accounting for the percentage of total bacteria count is 20%~40%；The percentage that the content of lactic acid bacteria accounts for total bacteria count is
20%~40%；The percentage that the ocean rhodotorula content accounts for total bacteria count is 10%~30%.
In a preferred embodiment, in an embodiment of the present invention, when described a kind of with arc in the inhibition shrimp pool
The leavening of the biological agent of bacterium growth includes bacteriophagic Bdellovibrio, Bdellovibrio starrii, bacillus, lactic acid bacteria, the red ferment in ocean
When female, the percentage that the bacteriophagic Bdellovibrio content accounts for total bacteria count is 10%；The Bdellovibrio starrii content accounts for total bacteria count
Percentage is 10%；The percentage that the bacillus content accounts for total bacteria count is 30%；The content of lactic acid bacteria accounts for total bacteria count
Percentage is 30%；The percentage that the ocean rhodotorula content accounts for total bacteria count is 20%.
It is understood that those skilled in the art can be according to strain formula provided by the invention, bacterium according to routine
Additive amount (mass percent of bacterium is 1~2%) is added corresponding culture medium and is mixed and made into bacterium solution or bacterium powder, is specifically commercially answering
In, what the present invention protected there is the leavening for inhibiting the biological agent of vibrios growth in the shrimp pool can be bacterium solution or bacterium powder.
In an embodiment of the present invention, the present invention provides a kind of with the biological agent of vibrios growth in the inhibition shrimp pool
Leavening, the leavening include the hair with the biological agent for inhibiting that vibrios grows in the shrimp pool that first aspect present invention provides
Ferment agent further includes fluid nutrient medium, solid medium or powdered medium.
In a preferred embodiment, the production medium of the compound bacteria A comprises the following components in parts by weight:
1000 parts of clean tap water, 100 parts of corn flour, 25 parts of bean cake powder, 0.03 part of manganese sulfate, 1 part of magnesium sulfate, sodium dihydrogen phosphate 1.5
Part, 3 parts of disodium hydrogen phosphate；Its optimal technological condition for fermentation is 28 DEG C of cultivation temperature, initial pH7.0, ventilatory capacity 2.0m3/ h, tank
Press 0.03MPa, fermentation period 36h.Bacterial concentration has reached 8.0 × 1010cfu/mL。
In the preferred embodiment of the present invention, the production medium of the compound bacteria B includes the group of following parts by weight
Point: 1000 parts of clean tap water, 12 parts of molasses, 5 parts of yeast extract, 1 part of peptone, 4 parts of glucose, 3 parts of corn pulp, sodium acetate 5
Part, 5 parts of sodium chloride, 2.5 parts of dipotassium hydrogen phosphate, 2.5 parts of potassium dihydrogen phosphate, 0.5 part of magnesium sulfate, 0.25 part of manganese sulfate.
Second aspect of the present invention provides a kind of biological agent with vibrios growth in the inhibition shrimp pool, mainly by the present invention
The activated post-fermentation raw material preparation gained of the leavening that first aspect provides, wherein the raw material includes that weight proportion is 1:1
Dregs of beans and wheat bran.
In an embodiment of the present invention, the biological agent, including compound bacteria A and compound bacteria B；
Wherein, the compound bacteria A includes one of bacteriophagic Bdellovibrio, Bdellovibrio starrii or a variety of；The compound bacteria B
Including one of bacillus, lactic acid bacteria, ocean rhodotorula or a variety of.
In an embodiment of the present invention, when described a kind of with the leavening for inhibiting the biological agent of vibrios growth in the shrimp pool
When including bacteriophagic Bdellovibrio, Bdellovibrio starrii, bacillus, lactic acid bacteria, ocean rhodotorula, the bacteriophagic Bdellovibrio content is accounted for
The percentage of total bacteria count is 5%~15%；The percentage that the Bdellovibrio starrii content accounts for total bacteria count is 5%~15%；Institute
Stating bacillus content and accounting for the percentage of total bacteria count is 20%~40%；The percentage that the content of lactic acid bacteria accounts for total bacteria count is
20%~40%；The percentage that the ocean rhodotorula content accounts for total bacteria count is 10%~30%.
In a preferred embodiment, when described a kind of with the hair for inhibiting the biological agent of vibrios growth in the shrimp pool
When ferment agent includes bacteriophagic Bdellovibrio, Bdellovibrio starrii, bacillus, lactic acid bacteria, ocean rhodotorula, the bacteriophagic Bdellovibrio contains
The percentage that amount accounts for total bacteria count is 10%；The percentage that the Bdellovibrio starrii content accounts for total bacteria count is 10%；The gemma
The percentage that bacillus content accounts for total bacteria count is 30%；The percentage that the content of lactic acid bacteria accounts for total bacteria count is 30%；The ocean
The percentage that rhodotorula content accounts for total bacteria count is 20%.
In an embodiment of the present invention, leavening activation activation medium used includes the group of following parts by weight
Point: 1000 parts of cultivating pool water, 20 parts of brown sugar, 0.5 part of ammonium sulfate, 0.05 part of epsom salt, 0.5 part of dipotassium hydrogen phosphate, carbon
14.29 parts of sour calcium, 2 parts of sodium chloride, 1 part of brine, 0.02 part of a water calcium chloride, 0.05 part of ferrous sulfate heptahydrate, cupric sulfate pentahydrate
0.05 part.
Third aspect present invention provides a kind of with the preparation method for inhibiting the biological agent of vibrios growth in the shrimp pool, packet
Include following steps:
(1) pretreatment of fermentation raw material: dregs of beans and wheat bran that weight proportion is 1:1 are crushed, the first mixing is obtained
Material；
(2) activation of leavening: compound bacteria A and compound bacteria B are added into activation medium respectively, must be fermented after culture activation
Liquid A and fermentation liquid B；
(3) it ferments: step (2) is added toward step (1) resulting first mixture and prepare resulting fermentation liquid A, obtain second
Mixture, aerobic fermentation are added fermentation liquid B, obtain third mixture, sealed fermenting.
(4) low molecule fulvic acid is added toward step (3) resulting third mixture, stirs and evenly mixs, can there must be inhibition
The biological agent that vibrios grows in the shrimp pool.
In an embodiment of the present invention, in the step (2), activation medium is comprised the following components in parts by weight: cultivation
1000 parts of pond water, 20 parts of brown sugar, 0.5 part of ammonium sulfate, 0.05 part of epsom salt, 0.5 part of dipotassium hydrogen phosphate, calcium carbonate
14.29 parts, 2 parts of sodium chloride, 1 part of brine, 0.02 part of a water calcium chloride, 0.05 part of ferrous sulfate heptahydrate, cupric sulfate pentahydrate 0.05
Part.
In an embodiment of the present invention, in the step (2), the activation culture temperature of compound bacteria A is 27-30 DEG C, culture
Time is 24-48 hours, is activated by the way of having oxygen activation.
In an embodiment of the present invention, in the step (2), the activation culture temperature of compound bacteria B is 27-30 DEG C, culture
Time is 24-48 hours, is activated by the way of sealing activation.
In an embodiment of the present invention, in the step (3), the weight proportion of the first mixture and fermentation liquid A are 1:2.
In an embodiment of the present invention, in the step (3), the time of the second mixture aerobic fermentation is 2-3 days.
In an embodiment of the present invention, in the step (3), the time of third mixture sealed fermenting is 5-7 days.
In an embodiment of the present invention, in the step (4), the additional amount of low molecule fulvic acid is the 1 of third mixture
~2%.
Fourth aspect present invention provides the biology with vibrios growth in the inhibition shrimp pool as described in the first aspect of the invention
The leavening of preparation, the biological agent or such as present invention with vibrios growth in the inhibition shrimp pool as described in respect of the second aspect of the invention
There is the preparation method for inhibiting the biological agent of vibrios growth in the shrimp pool to regulate and control to lead in aquaculture water quality described in the third aspect
Application in domain.
The present invention has the beneficial effect that
One, by reasonably combined various probiotics, using all kinds of fermentation raw materials, and meanwhile it is living by some biologies of fermentation generation
Sex factor can be used as effective carbon source in environment, and the probiotics in directive breeding environment can fully demonstrate hair used
Synergistic effect between yeast-like fungi kind, the reduction ammonia nitrogen harm of highly effective and safe, improves water body safety guarantee, improves the various physics and chemistry of water body
Index improves water quality and substrate.
Two, it is big to introduce amino acid, vitamin, microelement of the biological organic matter rich in easily absorbed and balance etc.
Active material and a large amount of beneficial viable bacterias are measured, various nutrition are complete, and proportion rationally, can obviously increase cultivation after applying on prawn culturing
Water body active organic matter content provides good organic debris for prawn, improves prawn immunity of organisms.
Three, whole simple process, the requirement to equipment is not also high, strong operability, easily in scale raising or
It is to be promoted the use of in aquaculture cooperative society.
Specific embodiment
The following is a preferred embodiment of the present invention, it is noted that for those skilled in the art
For, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also considered as
Protection scope of the present invention.
Below by a preferred embodiment of the present invention will be described in detail.The experiment of actual conditions is not specified in embodiment
Method, usually according to normal condition.In the embodiment of the present invention unless otherwise noted, agents useful for same and consumptive material are commercial goods.
Embodiment 1
1, the preparation of leavening
1) preparation of activation medium
Leavening activation activation medium used comprises the following components in parts by weight: 1000 parts of cultivating pool water, brown sugar
20 parts, 0.5 part of ammonium sulfate, 0.05 part of epsom salt, 0.5 part of dipotassium hydrogen phosphate, 14.29 parts of calcium carbonate, 2 parts of sodium chloride, halogen
1 part of water, 0.02 part of a water calcium chloride, 0.05 part of ferrous sulfate heptahydrate, 0.05 part of cupric sulfate pentahydrate.
2) compound bacteria A is activated:
1) addition prepares resulting activation medium by the way of having oxygen activation, 30 DEG C of cultures are activated for 36 hours,
Obtain fermentation liquid A.
3) activation of compound bacteria B:
1) addition prepares resulting activation medium by the way of sealing activation, 30 DEG C of cultures are activated for 36 hours,
Obtain fermentation liquid B.
2, with the preparation for inhibiting the biological agent of vibrios growth in the shrimp pool
1) pretreatment of fermentation raw material
By the raw material for needing to ferment, (each 50%) of dregs of beans, wheat bran is crushed, and obtains the first mixture；
2) it ferments
The fermentation liquid A of 2 times of quality is added toward resulting first mixture of step 1), stirs evenly, obtains the second mixture, 30
DEG C, aerobic fermentation 3 days, be then added with the fermentation liquid B of fermentation liquid equivalent, obtain third mixture, sealed fermenting 7 days.
3) 1.5% low molecule fulvic acid is added, stirs and evenly mixs, is in Umber pulp-water shape organic liquid, under have fermentation heavy
It forms sediment, stirring is in suspension, has fermenting aroma, as has the biological agent for inhibiting vibrios growth in the shrimp pool.Resulting biology system
10 times are diluted with the pool water when agent uses, full pool spilling head.
Embodiment 2
1, the preparation of leavening
1) preparation of activation medium
Leavening activation activation medium used comprises the following components in parts by weight: 1000 parts of cultivating pool water, brown sugar
20 parts, 0.5 part of ammonium sulfate, 0.05 part of epsom salt, 0.5 part of dipotassium hydrogen phosphate, 14.29 parts of calcium carbonate, 2 parts of sodium chloride, halogen
1 part of water, 0.02 part of a water calcium chloride, 0.05 part of ferrous sulfate heptahydrate, 0.05 part of cupric sulfate pentahydrate.
2) compound bacteria A is activated:
1) addition prepares resulting activation medium by the way of having oxygen activation, 27 DEG C of cultures are activated for 48 hours,
Obtain fermentation liquid A.
3) activation of compound bacteria B: 1) addition prepares resulting activation medium by the way of sealing activation, 27 DEG C of cultures
It is activated within 48 hours, obtains fermentation liquid B.
2, with the preparation for inhibiting the biological agent of vibrios growth in the shrimp pool
1) pretreatment of fermentation raw material
By the raw material for needing to ferment, (each 50%) of dregs of beans, wheat bran is crushed, and obtains the first mixture；
2) it ferments
The fermentation liquid A of 2 times of quality is added toward resulting first mixture of step 1), stirs evenly, obtains the second mixture, 30
DEG C, aerobic fermentation 3 days, be then added with the fermentation liquid B of fermentation liquid equivalent, obtain third mixture, sealed fermenting 7 days.
3) be added 2% low molecule fulvic acid, stir and evenly mix, be in Umber pulp-water shape organic liquid, under have fermentation precipitate,
Stirring is in suspension, has fermenting aroma, as has the biological agent for inhibiting vibrios growth in the shrimp pool.Resulting biological agent makes
Used time dilutes 15 times with pool water, full pool spilling head.
Embodiment 3
1, the preparation of leavening
1) preparation of activation medium
Leavening activation activation medium used comprises the following components in parts by weight: 1000 parts of cultivating pool water, brown sugar
20 parts, 0.5 part of ammonium sulfate, 0.05 part of epsom salt, 0.5 part of dipotassium hydrogen phosphate, 14.29 parts of calcium carbonate, 2 parts of sodium chloride, halogen
1 part of water, 0.02 part of a water calcium chloride, 0.05 part of ferrous sulfate heptahydrate, 0.05 part of cupric sulfate pentahydrate.
2) compound bacteria A is activated:
1) addition prepares resulting activation medium by the way of having oxygen activation, 30 DEG C of cultures are activated for 24 hours,
Obtain fermentation liquid A.
3) activation of compound bacteria B:
1) addition prepares resulting activation medium by the way of sealing activation, 30 DEG C of cultures are activated for 24 hours,
Obtain fermentation liquid B.
2, with the preparation for inhibiting the biological agent of vibrios growth in the shrimp pool
1) pretreatment of fermentation raw material
By the raw material for needing to ferment, (each 50%) of dregs of beans, wheat bran is crushed, and obtains the first mixture；
2) it ferments
The fermentation liquid A of 4 times of quality is added toward resulting first mixture of step 1), stirs evenly, obtains the second mixture, 30
DEG C, aerobic fermentation 2 days, be then added with the fermentation liquid B of fermentation liquid equivalent, obtain third mixture, sealed fermenting 5 days.
3) be added 1% low molecule fulvic acid, stir and evenly mix, be in Umber pulp-water shape organic liquid, under have fermentation precipitate,
Stirring is in suspension, has fermenting aroma, as has the biological agent for inhibiting vibrios growth in the shrimp pool.Resulting biological agent makes
Used time dilutes 20 times with pool water, full pool spilling head.
Embodiment 4
1, with the biological agent stability experiment for inhibiting vibrios growth in the shrimp pool:
After biological agent is kept in dark place 0 month, 2 months, 4 months at room temperature, in separately sampled activation detection microbial inoculum
Various bacterial contents, unit is (CFU/mL), as a result as follows:
Application Example
Application test of the biological agent provided by the invention in prawn culturing
1, test products: the biological agent of embodiment 1, total viable count is in 1,000,000,000/milliliter or more.
2, it tests prawn: throwing seedling to the prawn whole process formed and use.If 3 experiment pools and 3 control ponds.
3, test site: Guangdong Province Yangdong prawn culturing field.
4, test method: when daily raising, 1 biological agent of Example closes pond and uniformly splashes.It uses once, often within every five days
Mu breeding water body dosage is 1~2L, when water quality deterioration double using.And control group does not use biological agent.
5, test result:
The using effect for the biological agent that embodiment 1 provides, which is mainly reflected in, to be improved water quality, control water vibrios number and prevents
Shrimp disease etc. is controlled, specifically:
1) improve water quality, when water quality aging, composite bacteria agent of splashing, visible water colour is salubrious, tender, living within second day, validity period
Between, water colour is preferable, and pH stable, algal grown is vigorous, has no down that algae etc. occurs；
2) biological agent periodically provided using embodiment 1, water vibrios maintain reduced levels, and breeding process is substantially not
Use disinfectant；
3) cultivation is whole, and 3 experiment pool prawns of the biological agent provided using embodiment 1 have no disease substantially,
And often there is disease in 3 control ponds, using more disinfectant, occur because that disease seriously causes prawn massive mortality is final
Arrange the pool.When the final harvest of prawn, experiment pool increases production about 1000~1500kg/ mus than control pond.
The above embodiments are merely illustrative of the technical solutions of the present invention rather than limiting the scope of the invention, although ginseng
The present invention is explained in detail according to preferred embodiment, those skilled in the art should understand that, it can be to of the invention
Technical solution is modified or replaced equivalently, without departing from the spirit and scope of technical solution of the present invention.
Claims (10)
1. a kind of with the leavening for inhibiting the biological agent of vibrios growth in the shrimp pool, which is characterized in that including compound bacteria A and again
Combined bacteria B；Wherein, the compound bacteria A includes one of bacteriophagic Bdellovibrio, Bdellovibrio starrii or a variety of；The compound bacteria B packet
Include one of bacillus, lactic acid bacteria, ocean rhodotorula or a variety of.
2. leavening as described in claim 1, which is characterized in that bacteriophagic Bdellovibrio content accounts for total bacteria count in the leavening
Percentage is 5%~15%；The percentage that the Bdellovibrio starrii content accounts for total bacteria count is 5%~15%；The gemma bar
The percentage that bacterial content accounts for total bacteria count is 20%~40%；The content of lactic acid bacteria account for total bacteria count percentage be 20%~
40%；The percentage that the ocean rhodotorula content accounts for total bacteria count is 10%~30%.
3. a kind of with the biological agent for inhibiting vibrios growth in the shrimp pool, which is characterized in that mainly by as described in claim 1
The activated post-fermentation raw material preparation gained of leavening, wherein the raw material includes the dregs of beans and wheat bran that weight proportion is 1:1.
4. biological agent as claimed in claim 3, which is characterized in that leavening activation activation medium packet used
Include the component of following parts by weight: 800-1200 parts of cultivating pool water, 10-30 parts of brown sugar, 0.1-1 parts of ammonium sulfate, epsom salt
0.01-0.1 parts, 0.2-0.8 parts of dipotassium hydrogen phosphate, 10-20 parts of calcium carbonate, 1-3 parts of sodium chloride, 0.5-2 parts of brine, a water chlorination
0.01-0.05 parts of calcium, 0.01-0.1 parts of ferrous sulfate heptahydrate, 0.01-0.1 parts of cupric sulfate pentahydrate.
5. a kind of with the preparation method for inhibiting the biological agent of vibrios growth in the shrimp pool, which comprises the following steps:
(1) pretreatment of fermentation raw material: dregs of beans, wheat bran that weight ratio is 1:1 are crushed, the first mixture is obtained；
(2) activation of leavening: compound bacteria A and compound bacteria B are added into activation medium respectively, obtain fermentation liquid A after culture activation
And fermentation liquid B；
(3) it ferments: step (2) is added toward step (1) resulting first mixture and prepare resulting fermentation liquid A, obtain the second mixing
Material, aerobic fermentation are added fermentation liquid B, obtain third mixture, sealed fermenting；
(4) low molecule fulvic acid is added toward step (3) resulting third mixture, stirs and evenly mixs, can must have and inhibit the shrimp pool
The biological agent of middle vibrios growth.
6. preparation method as claimed in claim 5, which is characterized in that in the step (2), the activation culture temperature of compound bacteria A
Degree is 27-30 DEG C, and incubation time is 24-48 hours, is activated by the way of having oxygen activation.
7. preparation method as claimed in claim 5, which is characterized in that in the step (2), the activation culture temperature of compound bacteria B
Degree is 27-30 DEG C, and incubation time is 24-48 hours, is activated by the way of sealing activation.
8. preparation method as claimed in claim 5, which is characterized in that in the step (3), the first mixture, fermentation liquid A and
The weight proportion of fermentation liquid B is 1:2:2.
9. preparation method as claimed in claim 5, which is characterized in that in the step (3), the second mixture aerobic fermentation
Time is 2-3 days, and the time of third mixture sealed fermenting is 5-7 days.
10. the leavening with the biological agent for inhibiting that vibrios grows in the shrimp pool as described in claim 1, such as claim 3
The biological agent or such as claimed in claim 5 with vibrios in the inhibition shrimp pool with vibrios growth in the inhibition shrimp pool
Application of the preparation method of the biological agent of growth in aquaculture water quality regulation and control field.
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