CN1067720C - Bacterial strain for producing fishing microbial preparation and cultivating method thereof - Google Patents

Abstract

The present invention relates to a microbial preparation used for aquaculture animals. A Rhodopseudomonas CCTCC M96018 strain and a Lysobacter brunescens CTCC M96020 strain are used for culturing and preparing a lake-light preparation suitable for aquaculture animals in fresh water and brackish water; a Chromatium sp. CCTCC M96017 strain and a Flexibacter chimensis CCTCC M96019 strain are used for culturing and preparing a sea-light preparation suitable for aquaculture animals in brackish water and seawater. The preparation has the advantages of rich nutrients, fast growth and high stress resistance, and the effective period is as long as a half year when the preparation is stored under indoor natural light; the preparation performs obvious functions of promoting the growth and the development of various aquaculture animals, increasing the survival rate and the yield, improving the water quality and preventing diseases.

Description

The production method of fishing microbial preparation

The present invention relates to the preparation method of the microbial preparation that a kind of aquatic animal uses.

The most important condition of microbial preparation production is strain excellent and the optimum combination thereof that adapts to production requirement.In existing bibliographical information, microbial preparation relevant with photosynthetic bacterium or test belong to the single kind or the two or more mixed cultures of Rhodopseudomonas (Rhodopseudomonas), rhodospirillum (Rhodospirillum), Erythrobacillus (Rhodobacter) more with culture.These bacterial strains all belong to the Purple Non-sulfur photosynthetic bacterium, and are lower to sulfide tolerance in the environment to utilize less molecule organism, and general growth is all slower.More existing application tests show that some strain culture validity period is very short.

1、Pradal.M,1992.The?photosynthetic?bacteria?Rhodobactercapsulatus?as?the?food?complement?in?the?rainbow?trout?nutritioneffects?on?coloration?and?growth.ESGEC.313.

When France Pradal (1992) uses Rhodobacter capsulatus as rainbow trout, carp class additive of bait, must use prepared fresh, outmoded culture is then invalid.

2, Li Guangyou, Yu Yide, Jiang Yuxiang, fourth beauty, 1993. photosynthetic bacteriums are as the additive of bait test of prawn seed-rearing phase.The ocean material is learned (1): 52-54.

Li Guangyou etc. (1993) use Rhodopseudomonas bacterial strain (Rhodopseudomonas sp.) and find that in the prawn seed-rearing test cultivation person on the 7th is effective, surviving rate is significantly higher than control group (48.8%), 30 days persons only are higher than contrast 4%, and 45 days persons and contrast are as good as.

Light treasured (PSB) preparation that light precious Scientific and Technical Industry Co., Ltd in Hubei produces, the maintenance phase is 3 months, requires to preserve below 30 ℃.

Thailand produces crane and marks photosynthetic ferment (PSBIO) and at room temperature deposited less than 2 months, and assorted bacterium, assorted algae are many, require refrigeration, and resistance is poor, and validity period is short.These all are the limiting factors on this type of microbial preparation is used.

The objective of the invention is to the seed selection excellent species, and carry out optimum combination, very fast to obtain growth, adapt to fresh water, the different water bodys of seawater and culture object, be convenient to preserve the fishing microbial preparation that validity period is long.

For achieving the above object, its technical scheme characteristics are:

1, in conjunction with ecological Studies, by photosynthetic bacterium strain and the heterotrophic organism strain that sampling separates in the different water surroundings, seed selection has the production application potentiality.

2, on each bacterial strain biological characteristics basis of research, carry out the optimum combination of different strains, comprise that photosynthetic bacterium is with photosynthetic bacterial strain (photosynthetic bacterium has the bacterial strain of rhodopseudomonas, rhodospirillum, Chromatium); Photosynthetic bacterium is with the mixed culture of various combination between heterotrophic organism strain (heterotrophic organism has the dissolving bacillus, subdues the bacterial strain of genus such as bacillus).With the complementation of making the most of the advantage of different strains characteristic.CCTCC M96017-Chromatium (Chromafinm sp.) HQ strain; The mixed culture of CCTCC M96018-Rhodopseudomonas (Rhodopseudomonas) and rhodospirillum (Rhodospirillum), code name DL1-3.Bacillus (Flexibacterchinensis) FCA strain is subdued by CCTCC M96019-China.CCTCC M96020-browning dissolving bacillus (Lysobacter brunescens) M-4 bacterial strain.

Above bacterial strain is all delivered Chinese typical culture collection center (ChinaCenter for Type Culture Collection is called for short CCTCC) on October 30th, 1996, and detection finished on November 5th, 1996, proves all survivals.

3, be applicable to fresh water---brackish water respectively, brackish water by seed selection and combined preparation---two quasi-microorganism preparations of sea farming object and environment.

4, adopt three grades of training methods, guarantee the quality of the pharmaceutical preparations.Be first class inoculum, second class inoculum, three grades of enlarged culturing.

The present invention compared with prior art has following advantage and effect:

1, selected photosynthetic bacterium strain protein content height (60%), humans and animals indispensable amino acid are complete, and abundant carotenoid is arranged.

2, growth is very fast.The photosynthetic bacterium strain of selecting for use can reach 109/ml in 3-5 days under optimum conditions; When with different heterotrophic organism strain mixed culture, can improve its biomass 1.34-5.6 doubly; The mixed culture biomass is general equal between different photosynthetic bacterium strains improves 10%.

3, strong stress resistance, validity period is long.Lake-light, the used bacterial classification of sea light two class preparations were preserved respectively 6 years and 4 years at ambient temperature, still can survive.Preserve half a year---the preparation in 2 years also can be used as bait or does the additive of bait use, and general effective preservation period " lake-light " preparation 2 years lasts 15 days under 40 ℃ of dark conditions, still maintain vigour, and far the way is far away defeated; " sea light " preparation 1 year.

4, application effect.

Aquatic animal to grow such as A, promotion fish, shrimp are grown, and improve surviving rate, rate of body weight gain.The lake-light preparation that adds 1%-1.5% in the Macrobrachium rosenbergii breeding water body, its surviving rate generally all are higher than contrast 20%-25%, and the soprano reaches 4 times more than, and distortion ratio also is higher than contrast 22-49%.The sea light preparation is used for tigar prawn, japonicus and new prawn seed-rearing, and the highest seedling rate reaches 68.2%, 81.3% and 68.21% respectively, and shortens nursery stage 1-2 days.The Macrobrachium rosenbergii mature stage uses as additive of bait, improves output 21.5%, and individual weight in average is higher than control group 10.6%, and feed coefficient descends 22.2%.

In fresh water high-quality fingerling is raised, add 0.5% lake-light preparation in bait, grass carp kind day weight gain rate contrasts high by 36%, and the total augment weight rate is 216% of a control group.

B, promotion food organism propagation.When cultivating transparent flea, add 1-2% lake-light preparation, at 10 days internal breeding 90-100 doubly, contrast is then only bred 6-8.4 doubly.

C, improve water quality.After using this preparation, water colour is good, and water correction can reduce quantity of exchanged water and even not change water.After adding 6-7% immobilization preparation, in 96 hours, ammonia nitrogen, nitrite nitrogen and phosphoric acid salt descend 56.45%, 100% and 56.4% respectively in the water body, and the contrast water body rises 356.6%, 217% and 37.6% respectively.The effect that the prevention fish disease is arranged.

Implement method of the present invention and step:

Embodiment 1, and the lake-light preparation is applicable to fresh water and brackish water aquaculture water body.

A, lake-light preparation are produced bacterial strain and are formed through mixed culture by the rhodopseudomonas (Rhodopseudomonas sp.) of photosynthetic bacterium, the DL1-3 hybrid bacterial strain CCTCCM96018 and heterotrophic organism dissolving bacillus (Lysobacter brunescens) the M-4 bacterial strain CCTCCM96020 of rhodospirillum (Rhodospirillum sp.).

B, substratum:

The mixed culture of CCTCC M96018-Rhodopseudomonas (Rhodopseudomonas) and rhodospirillum (Rhodospirillum), code name DL1-3 bacterial strain RCVBN substratum.

CCTCC M96020-browning dissolving bacillus (Lysobacter brunescens) M-4 bacterial strain tryptone substratum, it is formed and proportioning is:

White peptone 0.1-1% of pancreas dawn

Yeast extract 0.1-0.5%

Beef extract 0.2-0.5%

Sodium-acetate 0.1-0.2%

pH 7-7.5

15 pounds of steam high-voltage sterilizings 20 minutes.

C, culture condition:

The mixed culture of CCTCC M96018-Rhodopseudomonas (Rhodopseudomonas) and rhodospirillum (Rhodospirillnm), code name DL1-3 bacterial strain requires illumination, illumination 1700-2000Lux, little good gas or anaerobism are cultivated, 30 ℃ ± 2 ℃ of temperature.

D, preparation production stage:

Get the mixed culture of CCTCC M96018-Rhodopseudomonas (Rhodopseudomonas) and rhodospirillum (Rhodospirillnm), code name DL1-3 bacterial strain, CCTCC M.96020-browning dissolving bacillus (Lysobacter brunescens) M-4 bacterial strain cultivates respectively, mixed culture with CCTCC M96018-Rhodopseudomonas (Rhodopsendomonas) and rhodospirillum (Rhodospirillum), code name DL1-3 inoculation is gone in the RCVBN substratum, 30 ℃ ± 2 ℃ illumination cultivation 72-120 hour.Get CCTCC M96020-browning dissolving bacillus (Lysobacter brunescens) M-4 inoculation and go in the tryptone substratum, cultivated 48 hours for 30 ℃ ± 2 ℃, then both cultures are connect 1: 1 mixed of capacity amplification cultivation again, results.

Embodiment 2, and the sea light preparation is applicable to brackish water and marine culture water.

A, preparation bacterial strain and culture condition:

Subdue bacillus (Flexibacter chinensis) FCA bacterial strain CCTCC M96019-China by the HQ hybrid bacterial strain CCTCC M96017 of photosynthetic bacterium Chromatium (Chromatium) and heterotrophic organism and subdue bacillus (Flexibacter chinensis) FCA strain, form through mixed culture.

CCTCC M96017-Chromatium (Chromatium sp.) HQ bacterial strain is cultivated with RCVBN.But require salinity, need increase the Nacl of 0.5-7% in the substratum.Illumination 1700-2000Lux, 30 ℃ ± 2 ℃ of temperature, anaerobism is cultivated.

Bacillus (Flexibacter chinensis) FCA strain bacterial strain is subdued by CCTCC M96019-China, uses the tryptone substratum, 30 ℃ ± 2 ℃ cultivations.

B, preparation production step gather:

Get CCTCC M96017-Chromatium (Chromatium sp.) HQ bacterial strain, CCTCC M96019-China subdues bacillus (Flexibacter chinensis) FCA strain bacterial strain and cultivates respectively, in the RCVBN substratum that increases 0.5-7%Nacl, 30 ℃ ± 2 ℃ illumination anaerobism were cultivated 72-120 hour with CCTCC M96017-Chromatium (Chromatium sp.) HQ inoculation.CCTCC M96019 China subdues bacillus (Flexibacter chinensis) FCA inoculation and goes in the tryptone substratum 30 ℃ ± 2 ℃ and cultivated 48 hours, then with 1: 1 by measure mixed amplification cultivation of both cultures, and results.

Application process, dosage: nursery stage can directly be sprayed in the nursery pond, presses the aquaculture water volume calculation, and amount of application is 5-10ppm, adds 5-10ml in the water promptly per ton.Time of application is preferably in the morning.Mature stage can be used with reference to aforesaid method dosage, also can add in the bait by the 0.5%-1% proportional and throw something and feed.Addition means can adopt to spray and be adsorbed on the granular bait for fish, perhaps adds in the Preblend, makes granular bait for fish then and uses.

Fishing microbial preparation is applicable to various aquatic animals, has the growth of promotion, grows, and improves surviving rate and output and improves water quality, and prophylactic effect also has effect same to the aquarium fish raising.This preparation is preserved under the room temperature natural light, and effectively the preservation period lake-light reaches 2 years; Sea light reaches 1 year, and nontoxic, has no side effect.

Claims (1)

1, the preparation method of fishing microbial preparation is characterized in that:

A, lake-light preparation, get CCTCC M96018 (mixed culture of Rhodopseudomonas Rhodopseudomonas and rhodospirillum Rhodospirillum) bacterial strain, CCTCCM96020 (browning dissolving bacillus Lysobacter brunescensv M-4) bacterial strain is cultivated respectively, CCTCCM96018 (mixed culture of Rhodopseudomonas Rhodopseudomonas and rhodospirillum Rhodospirillum) inoculation is gone in the RCVBN substratum 30 ℃ of illumination cultivation 72-120 of temperature hours.Getting CCTCC M96020 (browning dissolving bacillus Lysobacter brunescensvM-4) inoculation goes in the tryptone substratum, cultivated 48 hours for 30 ℃, then with 1: 1 by measure mixed amplification cultivation of both cultures;

B, the sea light preparation, get CCTCC M96017 (Chromatium Chromatium sp.HQ) bacterial strain, CCTCC M96019 (bacillus Flexibacterchinensis FCA subdues in China) bacterial strain is cultivated respectively, with CCTCC M96017 (Chromatium Chromatium sp.HQ) inoculation in the RCVBN substratum that increases 0.5-7%Nacl, 30 ℃ of illumination anaerobism were cultivated 72-120 hour, CCTCC M96019 (bacillus Flexibacter chinensis FCA subdues in China) inoculation is gone in the tryptone substratum 30 ℃ and was cultivated 48 hours, then 1: 1 by measure example of both cultures is mixed amplification cultivation.