CN104450579A - Microbial preparation for ecological breeding of penaeus vannamei boone and preparation method of microbial preparation - Google Patents

Abstract

The invention relates to a microbial preparation for ecological breeding of penaeus vannamei boone and a preparation method of the microbial preparation. The microbial preparation is formed by compounding bacillus subtilis with a water purification function, lactobacillus acidophilus and beneficial algae. The beneficial algae used in the invention comprise any one of cyanophyta, bacillariophyta, chlorophyta and chrysophyta.

Description

A kind of microbial preparation for Penaeus vannamei ecologic breeding and preparation method thereof

Technical field

The invention belongs to agricultural application microbial technology field.

Background technology

Shrimp culture industry occupies critical role in culture fishery.Shrimp culture industry once peaked in the early 1990s, but perplexed by disease afterwards, and output zhang width declines.In the world, the greatest danger that the development of prawn culture fishery and cultivation ability thereof face is day by day serious ecotope and pathologic problems, high-density breeding technology creation has gone out highly manual environment, this environment promotes bacterial growth, the selection of bacterium is evolved and creates disadvantageous effect: dissimilar water treatment, as mocromembrane filters and uviolizing, make seawater before being used to larya culturing, change microorganism species composition: the high-density young caused due to a large amount of Live food of throwing something and feeding, dead young remains, the high loading of organism and bacterium, have stimulated selective action, make the conditioned pathogen raised growth in aquaculture water.If now immunity of prawn weakens, disease will break out.The factors such as the continuous deterioration of shrimp disease and breeding environment, antibiotic abuse, irrational high-density breeding, prawn germ plasm resource is damaged, shrimp body disease resistance weakens have direct relation. and wherein environmental factors plays an important role.Disease of prawn break out be more cause disruption of ecological balance due to the destruction of the ecosystem and the increase of breeding density composite factor caused by.For shrimp culture industry, key improves breeding environment, improves prawn to the resistibility of disease.And probiotics both can purify water, environmental protect, prawn immunological competence can be improved again, there is good prospect in prawn culturing.

Summary of the invention

The object of this invention is to provide " a kind of microbial preparation for Penaeus vannamei ecologic breeding and preparation method thereof ", will there is the subtilis of water purification function, Lactobacterium acidophilum and the composite composition of useful algae.

Technical scheme

A kind of microbial preparation for Penaeus vannamei ecologic breeding of the present invention and preparation method thereof, will have the subtilis of water purification function, Lactobacterium acidophilum and the composite composition of useful algae; Useful algae in the present invention comprises, in Cyanophyta, Bacillariophyta, Chlorophyta, Chrysophyta 4 any one.

Its step is as follows:

(1) be inoculated in substratum by the inoculum size of 2%-10% by the subtilis activated and carry out shaking table cultivation, this substratum is: glucose 10-20g/l, peptone 2-5g/l, sodium-chlor 5-10g/l, PH=7.0-7.2.Culture condition is: 150-200r/min, temperature are 35-37 degree, air flow is 1:1; Subtilis viable bacteria pulvis is made after cultivating after 24-48 hour concentrated, drying;

(2) Lactobacillus acidophilus species activated is inoculated in substratum by the inoculum size of 5%-10% carries out shaking table cultivation, this substratum is as follows: molasses 20-30g/l, yeast extract paste 4-8g/l, potassium primary phosphate 5-10g/l, ammonium sulfate 10-20g/l, magnesium sulfate 0.5-2g/l, to regulate after PH=6-8 121 degree of sterilizings 20 minutes with tap water after dissolving; Culture condition is: culture temperature is that 30-37 degree quiescent culture is concentrated after 36-72 hour, live lactobacillus acidophilus pulvis is made in drying;

(3) by NaN03, KH2P04 and ironic citrate composite, the concentration of N, P and Fe is made to reach 10 ~ 14mg/L, 2 ~ 5mg/L and 0.14 ~ 0.3mg/L respectively, add pure water and put into 500mL Erlenmeyer flask, Erlenmeyer flask, through 120 degree of high-temperature sterilizations, is cooled to room temperature for subsequent use.Useful algae kind be inoculated in Erlenmeyer flask and cultivate, culture temperature is 25 ~ 35 DEG C, intensity of illumination 3000 ~ 4000Lx, and Light To Dark Ratio is 12:12, and salinity is 15, and every 3 h shake algae liquid once, and experimental period is 6 ~ 12 d.Then centrifugal, useful algae lyophilized powder is made in freeze-drying;

(4) by bacillus subtilis bacteria powder, live lactobacillus acidophilus pulvis, useful algae lyophilized powder is in (1 ~ 3): (1 ~ 3): the ratio of (1 ~ 3) carries out compositely namely obtaining microbial preparation.

Embodiment:

Embodiment 1:

(1) by the subtilis activated by 10% inoculum size be inoculated in substratum and carry out shaking table cultivation, this substratum is: glucose 20g/l, peptone 5g/l, sodium-chlor 10g/l, PH=7.0.Culture condition is: 200r/min, temperature are 35 degree, air flow is 1:1; Subtilis viable bacteria pulvis is made after cultivating after 24 hours concentrated, drying;

(2) by the Lactobacillus acidophilus species activated by 10% inoculum size be inoculated in substratum and carry out shaking table cultivation, this substratum is as follows: molasses 30g/l, yeast extract paste 8g/l, potassium primary phosphate 5g/l, ammonium sulfate 20g/l, magnesium sulfate 2g/l, to regulate after PH=6 121 degree of sterilizings 20 minutes with tap water after dissolving; Culture condition is: culture temperature is that 34 degree of quiescent culture are concentrated after 36 hours, live lactobacillus acidophilus pulvis is made in drying;

(3) by NaN03, KH2P04 and ironic citrate composite, make the concentration of N, P and Fe reach 14mg/L, 2mg/L and 0.14mg/L respectively, add pure water and put into 500mL Erlenmeyer flask, Erlenmeyer flask, through 120 degree of high-temperature sterilizations, is cooled to room temperature for subsequent use.Diatom algae kind be inoculated in Erlenmeyer flask and cultivate, culture temperature is 28 DEG C, intensity of illumination 4000Lx, and Light To Dark Ratio is 12:12, and salinity is 15, and every 3 h shake algae liquid once, and experimental period is 12 d.Then centrifugal, diatom lyophilized powder is made in freeze-drying;

(4) by bacillus subtilis bacteria powder, live lactobacillus acidophilus pulvis, diatom lyophilized powder carries out compositely namely obtaining microbial preparation in the ratio of 1:1:1.

Claims (3)

1. for a microbial preparation for Penaeus vannamei ecologic breeding, it is characterized in that: by subtilis, Lactobacterium acidophilum with useful algae is composite forms.

2. a kind of microbial preparation for Penaeus vannamei ecologic breeding according to claim 1, is characterized in that: useful algae comprises, in Cyanophyta, Bacillariophyta, Chlorophyta, Chrysophyta 4 any one.

3., for a preparation method for the microbial preparation of Penaeus vannamei ecologic breeding, it is characterized in that following steps:

(1) be inoculated in substratum by the inoculum size of 2%-10% by the subtilis activated and carry out shaking table cultivation, this substratum is: glucose 10-20g/l, peptone 2-5g/l, sodium-chlor 5-10g/l, PH=7.0-7.2;

Culture condition is: 150-200r/min, temperature are 35-37 degree, air flow is 1:1; Subtilis viable bacteria pulvis is made after cultivating after 24-48 hour concentrated, drying;

(2) Lactobacillus acidophilus species activated is inoculated in substratum by the inoculum size of 5%-10% carries out shaking table cultivation, this substratum is as follows: molasses 20-30g/l, yeast extract paste 4-8g/l, potassium primary phosphate 5-10g/l, ammonium sulfate 10-20g/l, magnesium sulfate 0.5-2g/l, to regulate after PH=6-8 121 degree of sterilizings 20 minutes with tap water after dissolving; Culture condition is: culture temperature is that 30-37 degree quiescent culture is concentrated after 36-72 hour, live lactobacillus acidophilus pulvis is made in drying;

(3) by NaN03, KH2P04 and ironic citrate composite, the concentration of N, P and Fe is made to reach 10 ~ 14mg/L, 2 ~ 5mg/L and 0.14 ~ 0.3mg/L respectively, add pure water and put into 500mL Erlenmeyer flask, Erlenmeyer flask, through 120 degree of high-temperature sterilizations, is cooled to room temperature for subsequent use;

Useful algae kind be inoculated in Erlenmeyer flask and cultivate, culture temperature is 25 ~ 35 DEG C, intensity of illumination 3000 ~ 4000Lx, and Light To Dark Ratio is 12:12, and salinity is 15, and every 3 h shake algae liquid once, and experimental period is 6 ~ 12 d;

Then centrifugal, useful algae lyophilized powder is made in freeze-drying;

(4) by bacillus subtilis bacteria powder, live lactobacillus acidophilus pulvis, useful algae lyophilized powder is in (1 ~ 3): (1 ~ 3): the ratio of (1 ~ 3) carries out compositely namely obtaining microbial preparation.