CN101760431B - A kind of compound microbial culture starter and application thereof - Google Patents

A kind of compound microbial culture starter and application thereof Download PDF

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Publication number
CN101760431B
CN101760431B CN201010033723.3A CN201010033723A CN101760431B CN 101760431 B CN101760431 B CN 101760431B CN 201010033723 A CN201010033723 A CN 201010033723A CN 101760431 B CN101760431 B CN 101760431B
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microbial inoculum
padding
application
yeast
numerous
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CN201010033723.3A
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CN101760431A (en
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王海宏
潘裕华
刘华贵
耿爱莲
张剑
初芹
肖银花
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北京市农林科学院
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Abstract

The present invention's " a kind of compound microbial culture starter and application thereof ", belongs to the cost degradation application of effective microorganism preparation.A kind of compound microbial culture starter, does living microorganism wherein comprise yeast, subtilis (Bacillus? subtilis), bacillus natto (Bacillus? and milk-acid bacteria natto), does is described yeast yeast saccharomyces cerevisiae (Saccharomyces? cerevisiae), does is described milk-acid bacteria Lactococcus (Lactococcus? lactis).Can be used for the aspects such as livestock and poultry cultivation bed, fodder additives, its feature is that reducing the cost of user on starter drops into, and effect stability.

Description

A kind of compound microbial culture starter and application thereof
Technical field
The present invention relates to the application of beneficial microorganism starter in livestock and poultry cultivation field, particularly a kind of compound microbial culture starter and application thereof.
Technical background
Microbial technique is widely used in livestock birds health cultivation field, adopts Tiny ecosystem engineering philosophy, filters out applicable different livestock and poultry needs beneficial microbe strain, through the technique such as composite, make probiotics, add in drinking-water or feed, play disease-resistant yield-increasing, deodorizing and improve the effect of meat.At present, compound is closed microbial preparation and is widely used in almost all livestock and poultry cultivations such as pig, ox, rabbit, dog, chicken, duck, goose, dove, fish, soft-shelled turtle, shrimp.
For complex microorganism role on livestock and poultry cultivation can sort out following some: (1) increase raising poultry nutritive, improve efficiency of feed utilization, improve the speed of growth.(2) obviously strengthen fowl poultry immune ability and disease resistance, improve surviving rate.(3) improve the quality of meat, improve the quality of products.(4) reproductive capability of dam fowl is strengthened. (5) can remove fecaluria stench, purifying ecological environment, non-environmental-pollution.
Along with cultivation scale and cultivation the increasing sharply of quantity, the environmental pollution that large-scale cultivation brings, plant's environmental degradation, disease take place frequently and to problems such as the grievous injuries of animal welfare, annoying vast breeding enterprise.Probiotics obtains special concern in disease-resistant, the effect improved in purifying aquaculture environment.To be exactly micro ecological technology apply very cleverly of aquaculture for the fermentation bed cultivation technology of rising in recent years.
The principle of microecological fermentation bed to raise pig technology uses peculiar microorganism to make fermentable bedding and padding by raw materials such as fermentation sawdusts, be layered in pigsty with certain thickness, pig all lives on this zymophyte bed at whole growth phase, its movement is degraded rapidly by the microorganism in bacterium bed, is digested, can make to be destitute of smell in pig house, non-flushingly wash colony house, do not cause environmental pollution, economize on resources.Beneficial microorganism environment in pig house, suppresses the breeding of harmful bacteria, strengthens the immunizing power of pig, reduces epidemic prevention medication, increases surviving rate and the rate of animals delivered to the slaughter-house, reach the object improving product quality and culture benefit.Bedding and padding in breeding process and ight soil directly generate high quality organic fertilizer, turn waste into wealth, additional income.
This aquaculture model at present, all more and more paid attention in countries in the world, the agriculture production expert of various places and researcher just drop into a large amount of work, the Tiny ecosystem that research and development are suitable for national this area is raised pigs pattern, apply through preliminary, all obtained certain success, the boundary that is subject to raising pigs is praised highly.Although this pattern is also in the early stage of development now, and development and improve in the middle of, its economy, the feature of environmental protection fully manifest.
Microecological fermentation bed cultural technique is due to advantages such as it is energy-conservation, reduction of discharging, environmental protection, safety, welfares, extensively approved by government and raiser, but because relevant supporting technology is not in place, input selling at exorbitant prices increases the factors such as application cost, significantly limit applying of fermentation bed cultivation technology.Current ecological fermentation bed cultural technique special leaven commercial prod is numerous, price is all higher, the starter cost that every square metre of bedding and padding increase does not reach 30-60 unit not etc., and quality product is uneven, also usually there is unstable phenomenon in effect, has a strong impact on the effect of fermentation bed cultivation and apply process.
Summary of the invention
According to the defect in above-mentioned field, the invention provides a kind of compound microbial culture starter and application thereof, this starter is formed by the assembly of four kinds of beneficial microorganism bacterium liquid, can be used for the aspects such as livestock and poultry cultivation bed, fodder additives, its feature is that reducing the cost of user on starter drops into, and effect stability.
A kind of compound microbial culture starter, living microorganism wherein comprises yeast, subtilis (Bacillussubtilis), bacillus natto (Bacillusnatto) and milk-acid bacteria, described yeast is yeast saccharomyces cerevisiae (Saccharomycescerevisiae), and described milk-acid bacteria is Lactococcus (Lactococcuslactis).
Described yeast is 0.01% access the former bacterium of yeast by volume in liquid medium within, cultivates 10 ~ 15 hours obtained bacterium liquid at 33 DEG C ~ 34 DEG C;
Described subtilis is the Bacillus subtilis strain liquid by 1% inoculation secondary enlarged culturing in liquid medium within, at 31 DEG C ~ 32 DEG C, cultivates the bacterium liquid of 30 ~ 35 hours gained;
Described bacillus natto is the bacillus natto strain liquid by 1% inoculation secondary enlarged culturing in liquid medium within, at 31 DEG C ~ 32 DEG C, cultivates the bacterium liquid of 30 ~ 35 hours gained;
Described milk-acid bacteria is the lactic acid bacteria culturers liquid by 1% access secondary enlarged culturing in liquid medium within, at 36 DEG C ~ 37 DEG C, and the bacterium liquid of static gas wave refrigerator 30 ~ 40 hours gained.
Bacterium liquid 3: 3: 3: 1 mixing by volume of described yeast, subtilis, bacillus natto, milk-acid bacteria four kinds of bacterium.
The application of described compound microbial culture starter in livestock and poultry cultivation is produced, it is characterized in that first carrying out numerous the obtaining of expansion to above-mentioned compound microbial culture starter expands numerous microbial inoculum, the numerous step of described expansion is as follows:
(1) brown sugar and water mixed and are stirred to abundant dissolving by weight at 1: 10, being cooled to 34 ~ 36 DEG C;
(2) add above-mentioned compound microecological starter 1 part, fully stir.Be placed in 30 ~ 35 DEG C of environment, within every 4 ~ 8 hours, stir once, cultivate 45 ~ 50 hours.
Described application refers to for livestock and fowl drinking water, it is characterized in that first 10 days that use, and adding by 1% in drinking-water and expand numerous microbial inoculum, can add-on be 0.3 ~ 0.5% after the 10th day.
Described application refers to that the numerous microbial inoculum of described expansion adds in complete feed by 0.5%, and described interpolation refers to mix thoroughly with complete diet pellet or spray on complete diet pellet surface.
Described application refers to expand numerous microbial inoculum by 0.1% dilution, sprays the breeding environment carrying out livestock and poultry as thimerosal livestock and poultry and carries out disinfection.
Described application refers to make fermented feed, according to feed: water: expand numerous microbial inoculum: brown sugar=100: the volume ratio of 60: 1: 1 is mixed thoroughly, load lucifuge fermentation in sealing bag and obtain described fermented feed in 5-7 days, by 10% of feeding capacity, fermented feed is joined in complete diet pellet, mix livestock and poultry of feeding thoroughly.
Described application refers to make fermenting bed padding, mass volume ratio 1kg: 5M of the numerous microbial inoculum of described expansion and bedding and padding 3, described bedding and padding and microbial inoculum mixed fermentation at normal temperature, be raised to for its feeding after more than 35 DEG C to bedding and padding internal temperature, and described microbial inoculum is that the numerous microbial inoculum of expansion and water are mixed into, and described microbial inoculum accounts for 30 ~ 40% of dunnage weight.
Described mixed fermentation first spreads the rice husk of 20 cm thicks, corn cob meal or/and roll short corn stalk at the circle end, spray microbial inoculum, repave the sawdust of 10 centimeters, rice husk or/and Pericarppium arachidis hypogaeae, spray microbial inoculum, dark to 80 centimetres with this paving of circulating, last one deck spreads 10 centimetres of sawdusts, surface spray microbial inoculum, then cover gunnysack or woven bag, treat that the lower 30 centimeters of temperature of bedding and padding are raised to more than 35 DEG C.
A kind of compound microbial culture starter provided by the invention, for livestock and poultry cultivation provides a kind of practical, efficient starter is the mixed bacteria liquid of four kinds of probioticss, the present invention, by contrast optimization experiment, have selected the bacillus natto of aerobic fermentation strong stress resistance, subtilis:
Bacillus natto has probiotics that is acidproof, heat-resistant quality, under hydrochloric acid in gastric juice, four hours survival are 100%, there is powerful pathogenic bacteria rejection ability simultaneously, in the middle of various beneficial bacterium, best to environment resistance, and one of bacterial classification of the small intestine that can go directly, animal intestinal flora can be changed ecological after oral, help digest function normalizing, to make defecation smooth and easy, maintain the effect of body physiological balance; Can produce acid, regulating intestinal canal flora, enhancing animal cell immunity is put should; And multiple protein enzyme particularly Sumizyme MP, saccharifying enzyme, lipase, amylase can be generated, the carbohydrate of some complexity in degrading plant feed, thus improve the transformation efficiency of feed.
Subtilis, can resistance to oxidation, anti-extrusion is high temperature resistant, can resistance to 60 DEG C of high temperature for a long time, to survive at 120 DEG C of temperature 20 minutes, acid and alkali-resistance, can keep active in acidic stomach environment, can the attack of resistance to saliva and bile, be can the viable bacteria of 100% through intestine and small intestine in feed microbe; The dominant population of high bacteria containing amount can be multiplied in a short time, promote the breeding of useful anaerobion, suppress the growth of unwanted bacteria (intestinal bacteria, Salmonella); In Fast-propagation process, produce a large amount of multivitamin, organic acid, amino acid, proteolytic enzyme particularly Sumizyme MP, saccharifying enzyme, lipase, amylase, organism complicated in energy degrading plant feed, thus promoting digestion absorbs, improve efficiency of feed utilization, prevent animal digestion bad, occur that situations such as " feed just " occurs; Fly is driven in deodorizing, decreasing pollution, controls bacteriosis, the excretion of nitrogen in ight soil, phosphorus, calcium can be reduced, reduce stool odor and noxious gas emission, show as animal excrement stink and progressively alleviate, reduce feed protein and be decomposed into ammonia waste, thus play the effect of cleaning ambient pollution.
Consider the dual-purpose of compound microbial culture starter of the present invention, to the prebiotic effect of animal and the factor for the potential of hydrogen that maintains bedding and padding during fermentation bed, present invention adds Lactococcus and yeast saccharomyces cerevisiae carries out compatibility symbiosis.Lactococcus is the bacterium of being everlasting in animal body, pass through biological antagonist in animal body, reduce pH value, stop and suppress intrusion and the field planting of pathogenic bacterium, degraded nitrous ammonia, ammonia, indoles, the objectionable impuritiess such as skatole, maintain the normal eubiosis in enteron aisle, higher SOD is contained in viable bacteria body and in meta-bolites, humoral immunization and cellular immunization can be strengthened, the Nisin that Lactococcus lactis produces is to suis, staphylococcus, some in genus bacillus are planted, clostridium and other milk-acid bacterias have restraining effect, to the potential of hydrogen maintaining fermentation bedding and padding, promote that growth of animal aspect effect is better than other milk-acid bacterias.Yeast saccharomyces cerevisiae selected by the present invention can well with Lactococcus symbiosis, to animal house deodorizing and more obvious to the probiotic effects of animal after compatibility, therapeutic E.coli K88, K99 and 987P can be suppressed, metabolism can produce immunostimulant, for animal provides foreign aid's property digestive tube probiotics, improve animal to the resistibility of disease.
Contained by this product, bacterial classification all belongs to the bacterial classification that can be applied to livestock-raising field of Ministry of Agriculture's regulation simultaneously, functional localization is clear and definite, with strong points, and application is simple, both can be used as fermentation bed special leaven, can be applied to again feed to add, through simultaneous test and productivity viewing test, it is high that this product is used for fermentation bed decomposition efficiency, improve obviously for fodder additives to animal productiong, improve animal resistibility, significantly reduce sickness rate, effect is stablized.
Present invention also offers a kind of starter that can reduce complex microorganism application cost in livestock and poultry cultivation, its feature is, wherein various raw material bacterium is and expands the lower bacterium liquid of numerous algebraically, such as, in starter of the present invention, yeast adopts the first-generation to expand numerous female bacterium bacterium liquid, other three kinds of bacterium all adopt the numerous bacterium liquid of expansion within the third generation, and the various raw material bacterium fermentation time of strict control and temperature, the object done like this is to make the bacterium in starter have more stable proliferation activity, because microorganism many generations expand numerous or expand numerous time oversize after, again with its be female bacterium expand numerous production microbial inoculum obtained, its activity and character are not difficult to keep efficient stable, and the microorganism that the present invention is to provide in starter expands at most numerous mistake twice, user can buy a small amount of starter of the present invention and carry out voluntarily fermenting expanding and numerously obtain a large amount of can directly use and the compound microbial culture starter that ensures of effect, and nutrient solution used is with low cost, such raiser spends the cost on microbial starter culture almost negligible.
In the preferred embodiments of the present invention, provide preferred fermentation time and the temperature of various raw material bacterium in compound microbial culture starter, inventor is by a series of simultaneous test, also show that the optimum blending ratio obtaining various raw material bacterium bacterium liquid under its preferred fermentation time above-mentioned and temperature is yeast: subtilis: bacillus natto: milk-acid bacteria four kinds of bacterium liquid by volume 3: 3: 3: 1, under this ratio, farthest can play the synergy between various bacterium, make compound microbial culture starter of the present invention to have taken into account decomposition efficiency high, improve the most aobvious for fodder additives to animal productiong, improve animal resistibility, remarkable reduction sickness rate, the advantages such as effect is stable.
The application method that the present invention also provides above-mentioned compound microbial culture starter to invent at livestock and poultry cultivation, simple and clear, economical, support the use with starter of the present invention, under the prerequisite of guarantee cultured product quality, aquaculture cost can be reduced for numerous raisers to the utmost.
The present invention still further provides the method for compound microbial culture starter of the present invention when the application of livestock and poultry cultivation each side, but the application of starter of the present invention is not limited to these methods.
Embodiment
The microbial strains that the present invention adopts and source thereof:
The former bacterium of yeast saccharomyces cerevisiae (SaccharomycescerevisiaeCICC31011 Sichuan Food Fermentative Industry Design Academy).
Subtilis (separation of fermentation industry Science Institute of BacillussubtilisCICC10036 first Ministry of Light Industry)
Bacillus natto bacterial strain (BacillusnattoCICC20132 Shanghai City Industry Wei Biological Research Institute (B167))
Lactococcus (key lab of the LactococcuslactisCICC22861 dairy science the Ministry of Education is separated (4.0610))
Above four bacterial strains, this experiment all has preservation, within 20 years from the applying date, can provide to the public, uses as research.
Embodiment 1
(1) cultivation of bacterial classification
1. cultivation method for saccharomycete
The preparation of nutrient solution: yeast extract paste 0.5%, peptone 0.5%, glucose 0.5%, produces in tank, heats to 90 DEG C, keeps 2 hours more than 90 DEG C.Then about 33 DEG C are cooled to.Access the former bacterium of 0.01% yeast saccharomyces cerevisiae (SaccharomycescerevisiaeCICC31011 Sichuan Food Fermentative Industry Design Academy).
Culture condition: at 33 DEG C ~ 34 DEG C, stir culture 12 hours.
It is stand-by that the yeast liquid produced puts into 4 DEG C of refrigerations.
2. subtilis cultural method
The cultivation of 2.1 bacterial classifications
2.1.1 the preparation of nutrient solution
Solid media for plates: extractum carnis 0.5%, yeast extract paste 0.5%, glucose 0.5%, peptone 1%, Nacl0.5%, agar 2%, adding distil water or deionized water dilution, PH7.2.
Liquid nutrient medium: soy peptone 2%, sucrose 2%, Sodium phosphate dibasic 0.2%, SODIUM PHOSPHATE, MONOBASIC 0.1%, manganous sulfate 0.05%, sodium-chlor 0.2%, adding distil water or deionized water dilution, put into triangular flask, 121 DEG C of autoclavings 20 minutes.Then about 30 DEG C are naturally cooled to.
2.1.2 the enlarged culturing of bacterial classification
Flat-plate solid one-level enlarged culturing: aseptically, get a fritter freeze-drying lactobacillus (separation of fermentation industry Science Institute of BacillussubtilisCICC10036 first Ministry of Light Industry) 0.5ml sterile purified water or deionized water dilution dissolving, then it is uniformly coated on solid medium, puts into more than 31 DEG C ~ 32 DEG C thermostat containers to 48 hour.Aseptically, the bacterial classification sterile purified water of one-level cultivation or deionized water flushing are put into sterile chamber stand-by.
Liquid two stage enlarged culturing: aseptically, the bacterium liquid that one-level is cultivated is accessed in liquid nutrient medium by 0.5%, and at 31 DEG C ~ 32 DEG C, 180 ~ 200rpm cultivates 33 hours.
2.2 produce the method with subtilis liquid
2.2.1 preparation 5% dregs of beans of nutrient solution, sucrose 2%, Semen Maydis powder 0.5%, sodium-chlor 0.2%, Sodium phosphate dibasic 0.2%, SODIUM PHOSPHATE, MONOBASIC 0.1, manganous sulfate 0.05%.With colloidal mill above-mentioned substance is milled to and is less than 80 order particles, put into and produce tank, heat to 90 DEG C, keep 2 hours more than 90 DEG C, be then cooled to about 31 DEG C.
2.2.2 inoculum size accesses 1% secondary enlarged culturing strain liquid in production tank.
2.2.3 at culture condition 31 DEG C ~ 32 DEG C, stir culture 33 hours.
It is stand-by that the bacillus liquid produced puts into 4 DEG C of refrigerations.
3. the cultural method of bacillus natto
The substratum of bacillus natto bacterial strain (BacillusnattoCICC20132 Shanghai City Industry Wei Biological Research Institute (B167)) bacillus natto: extractum carnis 0.5%, yeast extract paste 0.5%, glucose 0.5%, peptone 1%, Nacl0.5%, PH7.2
The cultural method of bacillus natto is with the production sequence of subtilis.
4. Lactococcus enlarged culturing method
The cultivation of 4.1 bacterial classifications
4.1.1 the preparation of nutrient solution
Solid media for plates: casein peptone 1%, beef extract 1%, yeast extract 0.5%, glucose 0.5%, sodium acetate 0.5%, dihydrogen citrate 0.2%, K2HPO40.2%, MgSO4.7H2O0.02%, MnSO4.H2O0.05%, 15% gram, agar, PH6.8.
Liquid nutrient medium: casein peptone 1%, beef extract 1%, yeast extract 0.5%, glucose 0.5%, sodium acetate 0.5%, dihydrogen citrate 0.2%, K2HPO40.2%, MgSO4.7H2O0.02%, MnSO4.H2O0.05%, PH6.8.Adding distil water or deionized water dilution, put into sealable bottle, 121 DEG C of autoclavings 20 minutes.Then about 36 DEG C are naturally cooled to.
4.1.2 the enlarged culturing of bacterial classification
Flat-plate solid one-level enlarged culturing aseptically, get a fritter freeze-drying lactobacillus (key lab of the LactococcuslactisCICC22861 dairy science the Ministry of Education is separated (4.0610)) 0.5ml sterile purified water or deionized water dilution dissolving, then it is uniformly coated on solid medium, puts into 36 DEG C ~ 37 DEG C anaerobic thermostat containers to 36 hour.Aseptically, the bacterial classification sterile purified water of one-level cultivation or deionized water flushing are put into sterile chamber stand-by.
Liquid two stage enlarged culturing aseptically, accesses the bacterium liquid that one-level is cultivated in liquid nutrient medium by 1%, container closure, at 36 DEG C ~ 37 DEG C, and Anaerobic culturel 36 hours.
The method of 4.2 production lactobacillus suspensions
4.2.1 the preparation Tryptones 1% of nutrient solution, yeast extract 0.5%, glucose 0.5%, sodium acetate 0.5%, dihydrogen citrate 0.2%, K2HPO40.2%, MgSO4.7H2O0.02%, MnSO4.H2O0.05%, PH6.8.Put into and produce tank, heat to 90 DEG C, keep 2 hours more than 90 DEG C, be then cooled to about 36 DEG C.
4.2.2 inoculum size accesses 1% secondary enlarged culturing strain liquid in production tank.
4.2.3 at culture condition 36 DEG C ~ 37 DEG C, static gas wave refrigerator 36 hours
It is stand-by that the bacillus liquid produced puts into 4 DEG C of refrigerations.
Embodiment 2 assembly compound microbial culture starter
Step 1. mixes bacterium liquid obtained in embodiment 1 by following volume ratio:
Subtilis: bacillus natto: yeast: milk-acid bacteria=3: 3: 3: 1 ratio mixing, add stablizer, coating-dividing sealing according to ten thousand/volume mass ratio, preserve under lucifuge condition.
Stablizer is SODIUM PHOSPHATE, MONOBASIC: Sodium phosphate dibasic in mass ratio 2: 1 mixed preparing obtains.
Step 2. Product checking
Yeast viable count>=10 8
Subtilis viable count>=10 7
Bacillus natto viable count>=10 7
Viable count of lactobacillus>=10 7
The application of embodiment 3, compound microbial culture starter
Step 1 expands numerous
1. get the container of more than 10 kilograms, as water vat, plastic tank, stainless steel cask, clean up, do not have any greasy, chemical residue.
2. 1 kilogram of brown sugar is poured in 10 kg of water, be stirred to and fully dissolve and boil, then naturally cool to about 35 DEG C, add the compound microbial culture starter 1 kilogram that embodiment 2 is obtained, after fully stirring, naturally cultivate.
3. in culturing process, envrionment temperature remains on 30 ~ 35 DEG C, within every 4 ~ 8 hours, stirs once, just becomes probiotics after 48 hours.
4., after cultured bacterium liquid stirs, available little mineral water bottle is scooped out one bottle and is left standstill observation, expands the good bacterium liquid of numerous effect, can occur skim white depositions after a few hours at the bottom of bottle.
(2) precaution
1. in culturing process, the best paper using of vessel port or gauze cover to be tightened, always not uncovered, to reduce other living contaminants.
2. temperature is low, and incubation time will lengthen, if envrionment temperature is lower than 22 DEG C, bacterial classification stops growing.
3. the microbial inoculum of probiotics stoste or enlarged culturing will avoid solar radiation, because the ultraviolet in sunlight can kill probiotic bacterium in preservation and culturing process.
4. cultured microbial inoculum was preferably finished in 3 ~ 7 days, and as cannotd be used up temporarily, should seal cryopreservation, preservation period does not exceed 30 days, and as occurred, stench does not use.User can need to determine to cultivate quantity according to the cultivation of oneself, long-time preservation of trying not.
5. probiotics stoste can only enlarged culturing once, can not many generations expand numerous, in order to avoid affect result of use.
(3) using method
The microbial inoculum of enlarged culturing can be used for the ecologic breeding of pig, chicken.Before using, fully shake up.
In the process using probiotic bacterium, feed and drinking-water do not add any microbiotic, if there is ill domestic animal, isolate for treatment separately.If need full group's medication, temporarily stop using probiotic bacterium, until drug withdrawal.
1. drink water: use for the first time, within first 10 days, drinking-water dosage can maintain 0.3 ~ 0.5% after 1%, 10 days.
2. spice: bacterium liquid adds in complete diet pellet by 0.5%, mixes thoroughly and feeds, the same day feeds, and the same day mixes.Also can spray with the direct charge level of diluent.
3. spray: by 0.1% dilution, carry out the sterilization of hen house band poultry as thimerosal, the effect of depositing dust and environment purification can be played.
4. make fermented feed: according to feed: water: bacterium liquid: brown sugar=100: all raw materials are mixed thoroughly by the ratio of 60: 1: 1, load in sealing bag, be placed in the place fermentation 5-7 days that lucifuge is warm, open bag sight material salubrious, without going mouldy, wine flavour is had to show to ferment successfully.Join in complete diet pellet by 10% of feeding capacity, mix thoroughly and feed.Fermented feed is finished behind Kaifeng as early as possible, is no more than two days winter, is no more than 1 day summer.
5. for fermenting bed padding: the bacterium liquid after 1 kilogram of expansion is numerous can make the bedding and padding of 5 cubes, need not additionally add the auxiliary materials such as Semen Maydis powder, wheat bran or brown sugar.Making method is as follows: adopt simple carvel built, first spreads rice husk or the corn cob meal of 20 centimeters at the circle end or rolls short corn stalk, sprays bacterium liquid and water, it repaves the sawdust of 10 centimeters or rice husk or Pericarppium arachidis hypogaeae, water spray and bacterium liquid, and do agitation as appropriate, make uniform moisture.By that analogy until be paved with circle body, 80 centimeters dark, and last one deck paving 10 centimeters, sawdust, water spray and bacterium liquid on it, stir and make moisture distribution even, it covers gunnysack or woven bag.After 3-7 days, the lower 30 centimeters of temperature of bedding and padding are raised to more than 35 DEG C and just can enter swine rearing.Note: expand numerous microbial inoculum and add that the gross weight of the water of spray is 30% ~ 40% of dunnage weight.
More than 6 usages can be used alone, and also can combinationally use.
Life-time service probiotic bacterium, can reach strengthening immunity, the generation of anti-bacteria disease, and it is extremely naughty to reduce, and improves breeding environment, reduces animal house stench, and by reducing cultivation medication, reaches the object improving meat quality and safety, thus improves culture benefit.
Embodiment 4 application example and effectiveness comparison
1 materials and methods
Select cotton stalk powder, sawdust, rice husk to be that main raw material carries out proportioning, make fermentation bed to raise pig bedding and padding
Microbial inoculum: choose market and sell good fermentation bed special bacteria agent 2 kinds, be designated No. 1, microbial inoculum (main bacteria seed consists of: lactobacillus, genus bacillus, yeast, photosynthetic bacteria), No. 2, microbial inoculum (main component is: yeast, milk-acid bacteria, photosynthetic bacteria, actinomycetes, mould), with the obtained compound microbial culture starter of the embodiment of the present invention 2 through the method for embodiment 3 expand numerous after the microbial preparation (No. 3 microbial inoculums) that obtains carry out comparative study.
The usage ratio of three kinds of microbial inoculums and method are with the method described by embodiment 3< using method >5.
Experimental animal: greatly enhance, grow white bi-crossbreeding 107, mean body weight 68 kilograms, feeding period 30 days
Test design: need 6 swinerys, the evaluation test of bacterial classification adopts completely randomized design, and 3 kinds of microbial inoculum × 2 are repeated, totally six colony houses.
Grouping sees the following form 1:
Numbering 1# 2# 3# 4# 5# 6#
Pig's head number 10 10 10 10 10 15
Microbial inoculum 1 1 2 2 3 3
2 animal rearings and bedding and padding management expectancy
Free blanking crib, free choice feeding, by hurdle record feed quantity every day.Automatic drinking bowl is freely drunk water.Every test pig all will carry out overbit mark, and list is only weighed.Dietary nutritional standard is consistent, and immune programme for children is identical.
3 testing indexs
Production performance index: day weight gain, day feed consumption, feed-weight ratio
To ferment padding bed operation conditions:
Temperature measuring: weekly respectively measure fermentation bed surface and degree of depth 10cm, 30cm, 50cm temperature, often enclose choose 5 points; Simultaneously test colony house internal and external temperature, humidity.
Bedding and padding water content: simultaneously gather with the sample of carbon-nitrogen ratio, adopts the National Standard Method measuring method of moisture " in the feed " to detect moisture content in bedding and padding.
Total plate count: simultaneously gather with the sample of carbon-nitrogen ratio, adopts blood cell plate counting process to measure total viable count pH value
4 data analyses
Adopt spss10.0 statistical analysis software, statistical treatment is carried out to the data that test gathers.
5 results of study and analysis
5.1 padding bed production performance researchs measure
Testing data shows, there is notable difference in the padding bed impact of the production performance on growing and fattening pigs that different microbial inoculum makes, study selected 3 kinds of commodity microbial inoculums, wherein No. 3 microbial inoculums have good production performance, its day weight gain and day feed consumption and feed-weight ratio index be all obviously better than other two kinds of bacterium, in table 2.Feed-weight ratio respectively than No. 1 and No. 2 bacterium reduce by 14.2% and 9.0%.
The fermentation bed that table 2 uses different fermentations agent to make is on the impact of pig production performance
The Changing Pattern of total viable count in 5.2 bedding and padding operational processs
Carried out tracking and measuring to the Changing Pattern of total viable count in bedding and padding various in breeding process, measuring method is blood cell plate counting.
Sampling method: according to the principle collected specimens of 5 point samplings in each column home, directly gets after 0-15 centimetre of top layer fully mix, according to quartering, gets 1 part of pack, be used in reference to target and measure.
Data analysis shows, and in table 3, in bedding and padding, total count presents the trend increased gradually along with the prolongation of feeding time, and on average every gram of bedding and padding increase 1.5-2.5 hundred million viable bacteria weekly.
Total viable count change detection value (hundred million/gram) in time in the bedding and padding that the agent of table 3 different fermentations makes
Data presentation: the Changing Pattern of the bedding and padding total viable count that different microbial inoculum makes is basically identical, but bacterium number there are differences, and microbial inoculum 3 bedding and padding viable bacteria content is apparently higher than microbial inoculum 2 and 1, and microbial inoculum 2 viable count is minimum.Illustrate that different microbial inoculum there are differences, microbial inoculum 3 activity is stronger, and reproductivity resistance is all obvious due to other two kinds of microbial inoculums.
The huge fluctuation of later stage total viable count is because the different testing error caused of measuring method, non-padding bed practical situation.The Changing Pattern of padding bed temperature in 5.3 bedding and padding breeding process
The padding bed each position each degree of depth bedding and padding temperature averages (DEG C) that table 4 different strain makes
Different the padding bed of microbial inoculum follows identical change rule, 1# and 3# microbial inoculum bed tempertaure measured value is basically identical, and No. 2 microbial inoculum bed temperatures are obviously on the low side, the full phase on average 2.7 degree on the low side.Illustrate that the making of different microbial inoculum is padding bed and truly have difference.Padding bed temperature accumulated temperature reflects the active degree of wherein microbial reproduction breeding, and the glycolysis ability of fermentation bed to swine excrement that bed temperature is higher is higher, and this conforms to the measurement result of total count in bedding and padding.Illustrate that the active degree of microbial inoculum of the present invention is higher, ight soil glycolysis ability is stronger.
The change of pH value in 5.4 bedding and padding application processes
Test has carried out tracking monitor to the change of bedding and padding pH value in bedding and padding breeding process, samples weekly, takes 1.0 grams in sample, be placed in 100ml beaker, add 40ml deionized water.Solution ph is measured with ThermoOrion acidometer after dipping 30min.Result is as follows:
The fermentation bed pH change that the different microbial inoculum of table 5 makes
Acid-basicity in the pH value reflection bedding and padding of bedding and padding, the acid too high too low Growth and Reproduction being all unfavorable for bacterium in bedding and padding, different strain combinations may form good symbiosis homeostasis in bedding and padding, maintains comparatively suitable acid-basicity, ensures self normal growth and breeding.
Data presentation from table 5, get rid of and measure or sampling error, the pH of about 60 days 3 kinds of microbial inoculums does not have notable difference, but along with the prolongation of feeding time, No. 2 microbial inoculum pH, apparently higher than No. 3 bacterium and No. 1 bacterium, reflect the glycolysis ability of No. 2 bacterium not as No. 3 bacterium.
The change of moisture content in 5.5 bedding and padding application processes
The water content of bedding and padding decides the success or not of padding bed application to a great extent, and water content is too low, microbial inoculum activity not only can be caused to reduce, and a large amount of dust can increase pig and suffers from respiratory tract disease.And moisture is too high, can be subject to serious suppression by bacterial reproduction again, bed surface supporting capacity reduces rapidly, finally causes padding bed comprehensively routed.Therefore definitely understand the appropriate aqueous amount that fermentation bed is run, the maintenance for bed surface is most important.
The padding bed change of soil water content (%) of table 6 different strain
Research finds, padding bed making initial moisture about 60%, along with the prolongation of feeding time, moisture in bedding and padding constantly evaporates, water content declines gradually, and after 4-5 week, water content maintains 40%-50% substantially, fecaluria moisture and the fermentation bed transpiring moisture of pig reach in a basic balance, and bed surface situation is better.
There is certain impact in the water content of different microbial inoculums on bedding and padding, under same stocking density, No. 2 bacterium groups have one to iris out the rising of existing moisture, illustrate that the bearing capacity of bedding and padding obviously declines, be in the edge of collapse, and No. 1 bacterium and No. 3 bacterium moisture more suitable, there is no the risk of dead bed.
Conclusion
Shown by experimental study, adopt the compound microbial culture starter of the present invention's development, not only can significantly improve the production performance of pig, and the normal fermentation state of fermenting bed padding can be maintained, ensure the efficient glycolysis of fecaluria, reduce maintenance and the use cost of bedding and padding.This tool in production application is of great significance.

Claims (8)

1. a compound microbial culture starter, living microorganism wherein comprise yeast, subtilis ( bacillussubtilis), bacillus natto ( bacillusnatto) and milk-acid bacteria, described yeast be yeast saccharomyces cerevisiae ( saccharomycescerevisiae), described milk-acid bacteria be Lactococcus ( lactococcuslactis), described yeast is 0.01% access the former bacterium of yeast by volume in liquid medium within, cultivates 10 ~ 15 hours obtained bacterium liquid at 33 DEG C ~ 34 DEG C;
Described subtilis is the Bacillus subtilis strain liquid by 1% inoculation secondary enlarged culturing in liquid medium within, at 31 DEG C ~ 32 DEG C, cultivates the bacterium liquid of 30 ~ 35 hours gained;
Described bacillus natto is the bacillus natto strain liquid by 1% inoculation secondary enlarged culturing in liquid medium within, at 31 DEG C ~ 32 DEG C, cultivates the bacterium liquid of 30 ~ 35 hours gained;
Described milk-acid bacteria is the lactic acid bacteria culturers liquid by 1% access secondary enlarged culturing in liquid medium within, at 36 DEG C ~ 37 DEG C, the bacterium liquid of static gas wave refrigerator 30 ~ 40 hours gained, the 3:3:3:1 mixing by volume of described yeast, subtilis, bacillus natto, milk-acid bacteria four kinds of bacterium liquid.
2. the application of compound microbial culture starter according to claim 1 in livestock and poultry cultivation is produced, it is characterized in that first carrying out numerous the obtaining of expansion to described compound microbial culture starter expands numerous microbial inoculum, the numerous step of described expansion is as follows:
(1) by brown sugar and water by weight 1:10 mix and be stirred to abundant dissolving, be cooled to 34 ~ 36 DEG C;
(2) add described compound microecological starter 1 part, fully stir, be placed in 30 ~ 35 DEG C of environment, within every 4 ~ 8 hours, stir once, cultivate 45 ~ 50 hours.
3. application according to claim 2, refers to for livestock and fowl drinking water, it is characterized in that first 10 days that use, and adding by 1% in drinking-water and expand numerous microbial inoculum, can add-on be 0.3 ~ 0.5% after the 10th day.
4. application according to claim 2, refers to that the numerous microbial inoculum of described expansion adds in complete diet pellet by 0.5%, described in add refer to complete diet pellet mix thoroughly or spray complete diet pellet surface.
5. application according to claim 2, refers to that the numerous microbial inoculum of described expansion is by 0.1% dilution, sprays and carries out disinfection in the breeding environment of livestock and poultry.
6. application according to claim 2, refer to according to feed: water: expand numerous microbial inoculum: the volume ratio of brown sugar=100:60:1:1 is mixed thoroughly, load lucifuge fermentation in sealing bag and obtain fermented feed in 5-7 days, by 10% of feeding capacity, described fermented feed is joined in complete diet pellet, mix livestock and poultry of feeding thoroughly.
7. application according to claim 2, refers to make fermenting bed padding, the mass volume ratio 1kg:5M of the numerous microbial inoculum of described expansion and bedding and padding raw material 3, described bedding and padding raw material and microbial inoculum mixed fermentation at normal temperature, be raised to for its feeding after more than 35 DEG C to bedding and padding stockpile internal temperature, and described microbial inoculum is that the numerous microbial inoculum of expansion and water are mixed into, and described microbial inoculum accounts for 30 ~ 40% of dunnage weight.
8. application according to claim 7, described mixed fermentation refers to first to spread at the circle end rice husk of 20 cm thicks, corn cob meal or/and roll short corn stalk, spray microbial inoculum, repave the sawdust of 10 centimeters, rice husk or/and Pericarppium arachidis hypogaeae, spray microbial inoculum, with this paving of circulating to 80 cm thicks, finally spread the sawdust of 10 cm thicks, surface spray microbial inoculum, then covers gunnysack or woven bag, treats that the lower 30 centimeters of temperature of bedding and padding are raised to more than 35 DEG C.
CN201010033723.3A 2010-01-05 2010-01-05 A kind of compound microbial culture starter and application thereof CN101760431B (en)

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