CN101088362B - Fish and shrimp phagostimulant of fermented product adhesion protein and its preparation process - Google Patents

Abstract

The fish and shrimp phagostimulant of fermented product and adsorbed protein is prepared through screening beneficial bacteria seed from cultivated animal body, culturing in composite culture medium to proliferate, concentrating and adding plant protein with phagostimulant effect. It is added into fish and shrimp feed to raise their feed intake and improve their digesting function. The production process has combined beneficial microbe fermenting technology and low temperature drying technology to ensure the fast growth of beneficial bacteria, the well preservation of metabolic product and the special cultivating function. The fish and shrimp phagostimulant is one kind of green product capable of replacing traditional phagostimulant and antioxidant.

Description

Fish and shrimp phagostimulant and preparation method that a kind of tunning adhesion protein is produced

Technical field

The present invention relates to fish and shrimp phagostimulant and preparation method that a kind of tunning adhesion protein is produced; Through in fish feed, adding; The peat-reek and the profitable strain that utilize microbial fermentation to produce improve aquatic livestock enteron aisle and digestive function; Improve feed intake and the daily gain of animal, belong to the agro-ecology product technique field.

Background technology

Holding time for pursuing the fastest culture efficiency and improving feed in the aquaculture now; Anti-oxidants such as a large amount of amino acid, betaine and ethoxyquinoline in feed, have been added; Objectively reached and improved the effect of cultured output and food calling, but made feed produce strong peculiar smell simultaneously, and betaine and anti-oxidant all are Chemical Manufacture at present; Enrichment in culturing the fish body is for breeding production has been buried serious potential safety hazard.

At present; Disclosed patent, publication number is CN92111616.0, is entitled as the mussel protein powder that Production by Microorganism Fermentation has the prawn luring function; It is as carrier with various plants albumen; Animal mussel slurry is continuously fermented through two warm solids by multiple microorganism as the important protein source, produces a kind of animal protein feed.Mainly be used for phagostimulant and the protein source of aquatic products (prawn, fish) and poultry, domestic animal (chicken, ox etc.).What it was selected for use is special high protein culture medium and fermentate, does not carry out high-temperature sterilization and microbiological indicator detection simultaneously in the sweat, and the animal safety of its production is difficult to guarantee.

Summary of the invention

The purpose of this invention is to provide fish and shrimp phagostimulant and preparation method that the tunning adhesion protein is produced; Can substitute the use of betaine, anti-oxidant in the feed fully, utilize the peat-reek of microbial fermentation generation, reach the effect of food calling; Adsorb special vegetable protein simultaneously; When the nutrition of breed fish body is provided, cover the peculiar smell in the feed, reach the culture efficiency of green safety.

For realizing above purpose; Technical scheme of the present invention provides the fish and shrimp phagostimulant that a kind of tunning adhesion protein is produced; It is characterized in that, formulated by following percentage by weight: Lactobacillus casei 30-40%, Bifidobacterium 30-40%, false gastral cavity Candida 20-40%.

The preparation method of the fish and shrimp phagostimulant that a kind of tunning adhesion protein is produced is characterized in that, adopts fluid enlargement culture, in concentration process, adopts cryogenic separation and dry technology, in concentrating, adds vegetable protein simultaneously, and its preparation process is:

The first step: the separation of probiotics, purifying and preparation

A. separate: getting chyme in the crucian enteron aisle of health is the bacterium source; Utilize the lactic acid bacteria culture medium to carry out strain separating; Be separated to two kinds of lactic acid bacterias; The lactic acid bacteria that this product adopts has two kinds; For: Lactobacillus casei and licheniformis (13123), Bifidobacterium (Bifidobacterium ATCC7469), in the prawn enteron aisle, utilize yeast extract wort agar culture medium to separate a kind of saccharomycete, the saccharomycete that the present invention adopts is maternity leave gastral cavity Candida (Candida utilis CBCC5638);

B. purifying: the Lactobacillus casei that separation is obtained adopts the lactic acid bacteria culture medium to rule, separate with licheniformis, Bifidobacterium (Bifidobacterium), and false gastral cavity Candida (Candida utilis) utilizes yeast extract wort agar culture medium to rule, separate;

C. the production production of hybrid seeds: the bacterial classification inoculation of purifying to the eggplant bottle that fills nutrient agar, was cultivated 28-30 hour under 30-35 ℃ of temperature, observed when lawn is yellow-white and is covered with eggplant bottle inclined-plane in the eggplant bottle, put into 4 ℃ of refrigerators and keep;

Second step: the Liquid Culture of bacterial classification

A. take by weighing Lactobacillus casei, Bifidobacterium and the false gastral cavity Candida of the step 1 production production of hybrid seeds by weight ratio, inoculating proportion of the present invention is: Lactobacillus casei 30-40%, Bifidobacterium 30-40%, false gastral cavity Candida 20-40%;

B. with the Lactobacillus casei that takes by weighing, Bifidobacterium, false gastral cavity Candida in 1: the ratio of 20-33 is then in 120-130 ℃, the complex medium of 30-40min sterilization; Put into fermentation tank then and stir 30min; Its rotating speed is 200r/min; Put into sterilization back Plastic Drum after stirring and left standstill 10-13 days, leave standstill in the process and to measure Ph value, fermentation ends when Ph arrival 3.0-4.0 every day;

The 3rd step: concentrate absorption with vegetable protein

A. after the fermentation ends, being the screw settling centrifuge of 2700-3200r/min through rotating speed earlier with zymotic fluid, is that the butterfly centrifugal machine of 6800-7300r/min concentrates through rotating speed again, and concentration time is 4-8 hour;

B. concentrate the back and collect centrifugal thing, add corn protein powder in the 1-3% ratio, the mixture after the interpolation stirs in conical agitator, and control moisture is at 50-65%;

The 4th step: dry and pulverizing

A. adopt the Clothoid type drying machine; Steam Heating is carried out drying, and the control temperature is at 40-45 ℃, time 60-80min in drying, and moisture is detected in the dry back of finishing; Require dried material moisture≤12%; Color and luster is faint yellow or yellowish-brown, makes dry deposits yields Maillard reaction, and smell fragrance is strong;

B. crushing screening: dried material is pulverized on pulverizer, and the material after the pulverizing is crossed 50 eye mesh screens, packs into behind the sieve to have in the bucket or bag of inner bag, tightens sack, fastens Sign Board, changes warehouse for finished product over to and stacks in order;

Described complex medium is at least three kinds of mixtures that sterilized water adds bean cake powder, urea, clear chaff, ammonium sulfate, dusty yeast, corn flour, molasses, potassium dihydrogen phosphate, sodium hydrogen phosphate, magnesium phosphate, magnesium sulfate again, and its weight percent proportioning is: bean cake powder 2-5%, molasses 9-15%, dusty yeast 2-5%, potassium dihydrogen phosphate 0-0.6%, sodium dihydrogen phosphate 0.5-0.8%, magnesium sulfate 0-0.1%, sterilized water 73.6-85.5%.

The bacterial classification that the present invention adopts mainly is lactic acid bacteria and saccharomycete; Normal flora has material impact to the health of its body in the cultivated animals body, and the microecological balance between they and host has guaranteed the eubolism of host animal, has improved host's immune disease-resistance power; Also abundant nutriments such as vitamin are provided simultaneously for growing of host; Therefore can improve microbial population of animal intestinal tract, promote that the probiotics preparation of animal health growth is generally paid attention to, development is rapid, and it has mainly acted on:

(1) suppresses the harmful intestinal tract microorganism; After probio gets into alimentary canal; A large amount of breedings form dominant microflora in alimentary canal, thereby suppress harmful microbe propagation; Olsson (1992) confirms can suppress Vibrio anguillarum from the isolated bacterium of turbot enteron aisle with experiment in vitro, and it has stronger adhesive capacity than Vibrio anguillarum in enteron aisle mucus;

(2) supplement the nutrients; Improve organism metabolism, reduce feed coefficient, just contain a large number of nutrients as the probiotics preparation of additive itself; Like multiple proteins, vitamin, trace element, ubiquinone etc.; Probio also can produce many digestive enzymeses simultaneously, assists feed digestion, improves feed efficiency;

(3) stimulating immune system improves immunity, is good immunoactivator as the probiotics preparation of feed addictive, excites immunologic function, enhancing body immunity and premunition through producing the nospecific immunity regulatory factor;

(4) after the bacterial classification of the present invention metabolism products such as profitable strain and enzyme, vitamin, polyphenoils that in liquid fermentation, produce high concentration get into and culture the fish body; Can in fish body enteron aisle, breed rapidly; Improve fish body microecological balance, improve the human body immunocompetence, realize the breed purpose of green safety.

The phagostimulant that this patent product can be used as aquiculture animal adds; Its addition is 1: 800-1000, the bacterial classification in this patent product all from the aquatic livestock enteron aisle seed selection cultivate and to form, be that feedstuff industry 813 bulletins allow to add bacterial classifications; Pass through fluid enlargement culture; Produce profitable strain and a large amount of metabolite of high concentration, in concentration process, adopt cryogenic separation and dry technology, guaranteed that profitable strain and enzyme, amino acid, coenzyme, vitamin fully preserves; In concentrating, add special vegetable protein simultaneously, nutrient protein in the palatability of raising product and the supplement feed.

The invention has the advantages that:

1. adopt beneficial microbe fermentation technique and low temperature drying technology to combine, both guaranteed the high-speed rapid growth of probiotics, guaranteed that again its metabolite keeps fully, guaranteed that to greatest extent product is to culturing the special efficacy of body;

2, used bacterial classification all screens from cultivated animals, guarantees its metabolism security.Lactic acid bacteria is cultured body enteron aisle propagation, keeps the animal body microecological balance, regulates body immunity, and saccharomycete decomposes feed, produces a large amount of single cell proteins, satisfies and cultures the bulk-growth needs;

3, the special fermenting aroma food calling that utilizes microbial fermentation and produced, fragrant odour is strong, lasting, need not add phagostimulant and anti-oxidant, has guaranteed the food security in the breeding process, has reduced aquaculture cost;

4, through adding special vegetable protein, increase the food calling effect of product, replenished the nutrient protein of feed simultaneously;

5, production process is few, and not high to equipment and technological requirement, use amount is few, is easy in aquaculture, promote.

The specific embodiment

Below in conjunction with embodiment the present invention is described further.

Embodiment 1

The first step: the separation of probiotics, purifying and preparation

A. separate: getting chyme in the crucian enteron aisle of health is the bacterium source; Utilize MRS lactic acid bacteria culture medium to carry out strain separating; Be separated to two kinds of lactic acid bacterias; The lactic acid bacteria that this product adopts has two kinds, for: Lactobacillus casei and licheniformis (13123), Bifidobacterium (BifidobacteriumATCC7469).In the prawn enteron aisle, utilize yeast extract wort agar culture medium to separate maternity leave gastral cavity yeast, (Candidautilis CBCC5638);

B. purifying: adopt the lactic acid bacteria culture medium to rule, separate with licheniformis, Bifidobacterium (Bifidobacterium) Lactobacillus casei that is separated to; False gastral cavity Candida (Candida utilis) utilizes yeast extract wort agar culture medium to rule, separate, and the bacterial classification colony characteristics that requirement separates is obvious, colonial morphology is neat;

C. the production production of hybrid seeds: the bacterial classification that will rule, separates is inoculated into respectively in the eggplant bottle that fills nutrient agar, cultivates 28 hours under 32 ℃ of temperature, observes when lawn is yellow-white and is covered with eggplant bottle inclined-plane in the eggplant bottle, puts into 4 ℃ of refrigerators and keeps;

MRS lactic acid bacteria culture medium: yeast extract 7.5g, peptone 7.5g, glucose) 10g, potassium dihydrogen phosphate 2g, the 100ml of Tomato juice, Tween (tween) 800.5ml, Disti distilled water) 900ml pH7.0.

Yeast extract wort agar culture medium: malt flour 3g, yeast extract 0.1g, agar 15-20g, distilled water 1000ml.

Nutrient agar: beef extract 3g, peptone 10g, sodium chloride 5g, agar 15-20g, water 1000ml, pH7.0-7.2.

These culture mediums all can be found in the relevant operation in laboratory book.

Second step: the Liquid Culture of bacterial classification

A. take by weighing each bacterial classification of the step 1 production production of hybrid seeds by following weight proportion: Lactobacillus casei 30%, Bifidobacterium 30% and false gastral cavity Candida 40%;

B. with the Lactobacillus casei that takes by weighing, Bifidobacterium, false gastral cavity Candida in 1: 20 ratio then in 121 ℃, the fluid nutrient medium of 30min sterilization; Put into fermentation tank and stir 30min; Its rotating speed is 200r/min; Plastic Drum left standstill 10 days after putting into the sterilization of 100L after stirring, and left standstill in the process to measure Ph value, fermentation ends when Ph arrives 3.0-4.0 every day;

Its weight percent proportioning of complex medium is: bean cake powder 2%, molasses 9%, dusty yeast 3%, potassium dihydrogen phosphate 0.2%, sodium dihydrogen phosphate 0.5%, sterilized water 85.3%;

The 3rd step: concentrate absorption with vegetable protein

A. after the fermentation ends, being the screw settling centrifuge of 2700r/min through rotating speed earlier with zymotic fluid, is that the butterfly centrifugal machine of 6800r/min concentrates through rotating speed again, and concentration time is 5 hours;

B. concentrate the back and collect centrifugal thing, add corn protein powder in 1% ratio, the mixture after the interpolation stirs in conical agitator, and control moisture is 50%;

The 4th step: dry and pulverizing

A. adopt the Clothoid type drying machine, Steam Heating is carried out drying, and the control temperature is at 40 ℃, time 60min in drying; Make the suitable Maillard reaction of dry deposits yields; Make product color be milk yellow, smell fragrance is strong, further improves palatability, moisture is detected in the dry back of finishing; Require dried material moisture≤12%, color and luster is faint yellow or yellowish-brown;

B. crushing screening: dried material is pulverized on pulverizer, and the material after the pulverizing is crossed 50 eye mesh screens, packs into behind the sieve to have in the bucket or bag of inner bag, tightens sack, fastens Sign Board, changes warehouse for finished product over to and stacks in order.

Embodiment 2

The first step: with embodiment 1;

Second step: the Liquid Culture of bacterial classification

A. take by weighing each bacterial classification of the step 1 production production of hybrid seeds by following weight proportion: Lactobacillus casei 40%, Bifidobacterium 40% and false gastral cavity Candida 20%;

B. with the Lactobacillus casei that takes by weighing, Bifidobacterium, false gastral cavity Candida in 1: 33 ratio then in 121 ℃, the liquid complex medium of 40min sterilization; Put into fermentation tank and stir 30min; Its rotating speed is 200r/min; Plastic Drum left standstill 13 days after putting into the sterilization of 100L after stirring, and left standstill in the process to measure Ph value, fermentation ends when Ph arrives 3.0-4.0 every day;

Complex medium weight percent proportioning is: bean cake powder 5%, molasses 15%, dusty yeast 5%, potassium dihydrogen phosphate 0.5%, sodium dihydrogen phosphate 0.8%, magnesium sulfate 0.1%, sterilized water 73.6%.

The 3rd step: concentrate absorption with vegetable protein:

A. after the fermentation ends, being the screw settling centrifuge of 3200r/min through rotating speed earlier with zymotic fluid, is that the butterfly centrifugal machine of 7300r/min concentrates through rotating speed again, and concentration time is 7 hours;

B. concentrate the back and collect centrifugal thing, add corn protein powder in 3% ratio, the mixture after the interpolation stirs in conical agitator, and control moisture is 65%;

The 4th step: dry and pulverizing

A. adopt the Clothoid type drying machine, Steam Heating is carried out drying, and the control temperature is at 45 ℃, time 80min in drying; Make the suitable Maillard reaction of dry deposits yields; Make product color be milk yellow, smell fragrance is strong, further improves palatability, moisture is detected in the dry back of finishing; Require dried material moisture≤12%, color and luster is faint yellow or yellowish-brown;

B. with embodiment 1;

Add in fish, the shrimp feed in 0.8-1.0 ‰ ratio during use and stir, this product needs to seal up for safekeeping in that dry ventilation is airtight, should use as early as possible behind the Kaifeng.This product uses the back to culture fish body food ration every day will obviously be increased; Daily gain obviously improves, and in the popularization experiment that carry out at Spring in Pig House of Shanghai Fengxian aquatic products technical advice station, the per mu yield on each pool of test pond is 151.8 jin; And the average product on the contrast pool is merely 70 jin; The output of test pond has improved 116.9%, and difference is quite obvious, and feed coefficient has on average reduced by 17.1%.And the individuality of each test pond freshwater shrimp is bigger, body surface smooth, and body colour is blue or green, and selling price is higher, and each pool freshwater shrimp body surface of the contrast pool is crude, drift is more, and body colour is whiter, and selling price is not high.

Claims (1)

1. the preparation method of the fish and shrimp phagostimulant produced of a tunning adhesion protein, its preparation process is: the first step: the separation of probiotics, purifying and preparation

A. separate: in the crucian enteron aisle of health, getting chyme is the bacterium source; Utilize the lactic acid bacteria culture medium to carry out strain separating, be separated to two kinds of lactic acid bacterias, for: Lactobacillus casei and Bifidobacterium; Utilize yeast extract wort agar culture medium to separate a kind of saccharomycete in the prawn enteron aisle, for producing the gastral cavity Candida;

B. purifying: the Lactobacillus casei that separation is obtained adopts the lactic acid bacteria culture medium to rule, separate with Bifidobacterium, produces the gastral cavity Candida and utilizes yeast extract wort agar culture medium to rule, separate;

C. the production production of hybrid seeds: the bacterial classification inoculation that will rule, separates was cultivated 28-30 hour under 30-35 ℃ of temperature to the eggplant bottle that fills nutrient agar, observed when lawn is yellow-white and is covered with eggplant bottle inclined-plane in the eggplant bottle, put into 4 ℃ of refrigerators and kept;

Second step: the Liquid Culture of bacterial classification

A. take by weighing Lactobacillus casei, Bifidobacterium and the product gastral cavity Candida of the first step production production of hybrid seeds by weight ratio, proportioning is: Lactobacillus casei 30-40%, Bifidobacterium 30-40%, product gastral cavity Candida 20-40%;

B. with the Lactobacillus casei that takes by weighing, Bifidobacterium, product gastral cavity Candida in 1: the ratio of 20-33 is inoculated in 120-130 ℃, the complex medium of 30-40min sterilization; Put into fermentation tank then and stir 30min; Its rotating speed is 200r/min; Put into sterilization back Plastic Drum after stirring and left standstill 10-13 days, leave standstill in the process and to measure pH value, fermentation ends when pH arrival 3.0-4.0 every day; Described complex medium weight percent proportioning is: bean cake powder 5%, molasses 15%, dusty yeast 5%, potassium dihydrogen phosphate 0.5%, sodium dihydrogen phosphate 0.8%, magnesium sulfate 0.1%, sterilized water 73.6%;

The 3rd step: concentrate the absorption with vegetable protein, adopt cryogenic separation in the concentration process

A. after the fermentation ends, being the screw settling centrifuge of 2700-3200r/min through rotating speed earlier with zymotic fluid, is that the butterfly centrifugal machine of 6800-7300r/min concentrates through rotating speed again, and concentration time is 4-8 hour;

B. concentrate the back and collect centrifugal thing, add corn protein powder in the 1-3% ratio, the mixture after the interpolation stirs in conical agitator, and control moisture is at 50-65%;

The 4th step: dry and pulverizing

A. adopt the Clothoid type drying machine; Steam Heating is carried out drying, and the control temperature is at 40-45 ℃, time 60-80min in drying, and moisture is detected in the dry back of finishing; Require dried material moisture≤12%; Color and luster is faint yellow or yellowish-brown, makes dry deposits yields Maillard reaction, and smell fragrance is strong;

B. crushing screening: dried material is pulverized on pulverizer, and the material after the pulverizing is crossed 50 eye mesh screens, packs into behind the sieve to have in the bucket or bag of inner bag, tightens sack, fastens Sign Board, changes warehouse for finished product over to and stacks in order.