CN101760431A - Compound microbial culture starter and application thereof - Google Patents
Compound microbial culture starter and application thereof Download PDFInfo
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Abstract
The invention discloses a compound microbial culture starter and application thereof, which belong to an application of a beneficial microbial preparation at low cost. The active microorganism of the compound microbial culture starter comprises saccharmyces cerevisiae, bacillus subtilis, bacillus natto and lactococcus lactis. The compound microbial culture starter can be used for livestock and poultry raising beds, feeds addictive and other aspects and is characterized by reducing the cost input of users on the culture starter and having stable effect.
Description
Technical field
The present invention relates to the application of beneficial microorganism starter in the livestock and poultry cultivation field, particularly a kind of compound microbial culture starter and application thereof.
Technical background
Microbial technique is widely used in the livestock birds health cultivation field, adopts little ecological engineering principle, filters out to be fit to different livestock and poultry needs beneficial microbe strains, technologies such as process is composite, make probiotics, add in drinking-water or the feed, play disease-resistant yield-increasing, deodorizing and improve the effect of meat.At present, the compound microbial preparation that closes 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) increases raising poultry nutritive, improves efficiency of feed utilization, improves 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 enhancing dam fowl. (5) can remove the fecaluria stench, purifying ecological environment, non-environmental-pollution.
Along with the breed scale with culture increasing sharply of quantity, the environmental pollution that large-scale cultivation brought, plant's environmental degradation, disease takes place frequently and to the problems such as grievous injury of animal welfare, perplexing vast breeding enterprise.Probiotics has obtained special concern in effect disease-resistant, that improve aspect the purifying aquaculture environment.The fermentation bed cultivation technology of rising in recent years is exactly the very cleverly application of little ecotechnology in aquaculture.
The principle of microecological fermentation bed to raise pig technology is that the utilization peculiar microorganism is made the microbial fermentation bedding and padding by raw materials such as fermentation sawdusts, be layered in the pigsty with certain thickness, pig all lived on this zymophyte bed in the whole growth stage, its movement is degraded rapidly, is digested by the microorganism in the bacterium bed, can make in the pig house and be destitute of smell, the non-flushing colony house of washing does not cause environmental pollution, economizes on resources.Beneficial microorganism environment in the pig house suppresses the breeding of harmful bacterium, strengthens the immunizing power of pig, reduces the epidemic prevention medication, increases the surviving rate and the rate of animals delivered to the slaughter-house, reaches the purpose that improves product quality and culture benefit.Bedding and padding in the breeding process and ight soil directly generate high-quality fertilizer, turn waste into wealth additional income.
Present this aquaculture model, all more and more paid attention in countries in the world, the agriculture production expert and the researcher of various places just drop into a large amount of work, research and development are suitable for little ecologically raising pigs pattern of national this area, through preliminary applying, all obtained certain success, the boundary that is subjected to raising pigs is praised highly.Though this pattern also is in the early stage of development now, and in the middle of constantly developing and improving, its economy, the feature of environmental protection fully manifest.
Advantages such as microecological fermentation bed cultural technique is energy-conservation owing to it, reduction of discharging, environmental protection, safety, welfare, extensively approved by government and raiser, but because relevant supporting technology is not in place, the input selling at exorbitant prices increases factors such as application cost, has limited applying of fermentation bed cultivation technology greatly.Present ecological fermentation bed cultural technique special leaven commercial prod is numerous, price is all higher, the starter cost of every square metre of bedding and padding increase reaches 30-60 unit not to be waited, and quality product is uneven, also usually there is unsettled phenomenon in effect, has a strong impact on the effect of fermentation bed cultivation and applies process.
Summary of the invention
Defective according to 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 aspects such as livestock and poultry cultivation bed, fodder additives, its characteristics have been to reduce the cost input of user on starter, and effect stability.
A kind of compound microbial culture starter, living microorganism wherein comprises yeast, subtilis (Bacillus subtilis), bacillus natto (Bacillus natto) and milk-acid bacteria, described yeast is yeast saccharomyces cerevisiae (Saccharomyces cerevisiae), and described milk-acid bacteria is lactic acid coccus (Lactococcuslactis).
Described yeast is 0.01% to insert the former bacterium of yeast by volume in the liquid medium within, cultivates down for 33 ℃～34 ℃ and makes bacterium liquid in 10～15 hours;
Described subtilis be in the liquid medium within by the Bacillus subtilis strain liquid of 1% inoculation secondary enlarged culturing, under 31 ℃～32 ℃, cultivate the bacterium liquid of 30～35 hours gained;
Described bacillus natto be in the liquid medium within by the bacillus natto strain liquid of 1% inoculation secondary enlarged culturing, under 31 ℃～32 ℃, cultivate the bacterium liquid of 30～35 hours gained;
Described milk-acid bacteria be in the liquid medium within by the 1% lactic acid bacteria culturers liquid that inserts the secondary enlarged culturing, under 36 ℃～37 ℃, the bacterium liquid of 30～40 hours gained of static cultivation.
The bacterium liquid of described yeast, subtilis, bacillus natto, four kinds of bacterium of milk-acid bacteria 3: 3: 3 by volume: 1 mixes.
The application of described compound microbial culture starter in livestock and poultry cultivation is produced is characterized in that earlier above-mentioned compound microbial culture starter being expanded numerous numerous microbial inoculum that obtains expanding, and the numerous step of described expansion is as follows:
(1) brown sugar was mixed than 1: 10 by weight with water and be stirred to abundant dissolving, be cooled to 34～36 ℃;
(2) add 1 part of above-mentioned compound microecological starter, fully stir.Place 30～35 ℃ of environment, stirred once, cultivated 45～50 hours in per 4～8 hours.
Described application refers to be used for livestock and fowl drinking water, it is characterized in that using preceding 10 days, add by 1% in drinking-water and expand numerous microbial inoculum, but add-on is 0.3～0.5% after the 10th day.
Described application refers to that the numerous microbial inoculum of described expansion adds in the complete feed by 0.5%, and described interpolation refers to mix thoroughly or spray on the complete diet pellet surface with complete diet pellet.
Described application refers to expand numerous microbial inoculum by 0.1% dilution, sprays livestock and poultry to carry out disinfection as the breeding environment that thimerosal carries out livestock and poultry.
Described application refers to make fermented feed, according to feed: water: expand numerous microbial inoculum: brown sugar=100: 60: 1: 1 volume ratio is mixed thoroughly, packing into, the lucifuge fermentation obtained described fermented feed in 5-7 days in the sealing bag, by 10% of feeding capacity fermented feed was joined in the complete diet pellet, mixed the livestock and poultry of feeding thoroughly.
Described application refers to make fermenting bed padding, the mass volume ratio 1kg of numerous microbial inoculum of described expansion and bedding and padding: 5M
3, described bedding and padding and microbial inoculum mixed fermentation at normal temperature are used for its feeding after the bedding and padding internal temperature is raised to more than 35 ℃, and described microbial inoculum is mixed into for expanding numerous microbial inoculum and water, and described microbial inoculum accounts for 30～40% of dunnage weight.
Described mixed fermentation is spread the rice husk, corn cob meal of 20 cm thicks earlier or/and roll short corn stalk at the circle end, spray microbial inoculum, the sawdust, rice husk that repave 10 centimeters are or/and Pericarppium arachidis hypogaeae, the spray microbial inoculum, with this circulate the shop to 80 centimetres dark, the 10 centimetres of sawdusts in last one deck shop, surface spray microbial inoculum, cover gunnysack or woven bag then, treat that the following 30 centimeters temperature of bedding and padding are raised to more than 35 ℃ to get final product.
A kind of compound microbial culture starter provided by the invention, for livestock and poultry cultivation provides a kind of practicality, starter is the mixed bacteria liquid of four kinds of probioticss efficiently, the present invention has selected bacillus natto, the subtilis of aerobic fermentation strong stress resistance by the contrast optimization experiment:
That bacillus natto has is acidproof, the probiotics of heat-resistant quality, four hours survival rates are 100% under hydrochloric acid in gastric juice, have powerful pathogenic bacteria simultaneously and suppress ability, be in the middle of the various beneficial bacterium, best to the environment tolerance, and one of the bacterial classification of small intestine that can go directly, can change animal intestinal flora ecology after oral, the function that helps digest normalizing so that defecation is smooth and easy, is kept the effect of body physiological equilibrated; Can produce acid, regulate intestinal microflora, the enhancing animal cell immunity is put should; And can generate multiple protein enzyme particularly Sumizyme MP, saccharifying enzyme, lipase, amylase, some complicated carbohydrate in the degrading plant forage, thereby the transformation efficiency of raising feed.
Subtilis can resistance to oxidation, anti-extruding is high temperature resistant, can long-term anti-60 ℃ of high temperature, under 120 ℃ of temperature, can survive 20 minutes, acid and alkali-resistance, in the acid stomach environment, can keep active, can anti-saliva and biliary attack, be can 100% through intestine and small intestine in the feed microbe viable bacteria; Can breed into the dominant population of high bacteria containing amount in a short time, promote the breeding of useful anaerobion, suppress the growth of unwanted bacteria (intestinal bacteria, Salmonella); In quick reproductive process, produce a large amount of multivitamins, organic acid, amino acid, proteolytic enzyme particularly Sumizyme MP, saccharifying enzyme, lipase, amylase, complicated organism in the energy degrading plant forage, thereby promoting digestion absorbs, improve efficiency of feed utilization, prevent that animal digestion is bad, situations such as " feed just " occurs and take place; Fly is driven in deodorizing, reduces and pollutes, the control bacteriosis, can reduce the excretion of nitrogen in the ight soil, phosphorus, calcium, reduce ight soil stink and noxious gas emission, show as the animal excrement stink and progressively alleviate, reduce feed protein and be decomposed into the ammonia waste, thereby play the effect that cleaning ambient pollutes.
Consider the dual-purpose of compound microbial culture starter of the present invention, keep the factor of the potential of hydrogen of bedding and padding to the prebiotic effect of animal and when being used to ferment bed, the present invention has added the lactic acid coccus and yeast saccharomyces cerevisiae carries out the compatibility symbiosis.The lactic acid coccus is the intravital bacterium of being everlasting of animal, pass through biological antagonist in animal body, reduce the pH value, stop and suppress intrusion and the field planting of pathogenic bacterium, degraded nitrous ammonia, ammonia, indoles, objectionable impuritiess such as skatole, keep the normal eubiosis in the enteron aisle, contain higher SOD in the viable bacteria body and in the meta-bolites, can strengthen humoral immunization and cellular immunization, the Nisin that Lactococcus lactis produces is to suis, staphylococcus, some kinds in the genus bacillus, clostridium and other milk-acid bacterias have restraining effect, to keeping the potential of hydrogen of fermentation bedding and padding, promote growth of animal aspect effect to be better than other milk-acid bacterias.The selected yeast saccharomyces cerevisiae of the present invention can be well and the symbiosis of lactic acid coccus, behind the compatibility to animal house deodorizing and more obvious to the benefit of the animal fruit of coming into force, can suppress curing the disease property intestinal bacteria K88, K99 and 987P, can metabolism produce immunostimulant, for animal provides foreign aid's property digestive tube probiotics, improve the resistibility of animal to disease.
The contained bacterial classification of this product all belongs to the bacterial classification that can be applied to the livestock-raising field of Ministry of Agriculture's regulation simultaneously, functional localization is clear and definite, and is with strong points, uses simple, both can be used as fermentation bed special leaven, can be applied to feed again and add, through simultaneous test and productivity viewing test, this product is used for fermentation bed decomposition efficiency height, be used for fodder additives animal is produced improvement obviously, improve the animal resistibility, significantly reduce sickness rate, effect is stable.
The present invention also provides a kind of starter that can reduce complex microorganism application cost in the livestock and poultry cultivation, its characteristics are, wherein various raw material bacterium are expands the lower bacterium liquid of numerous algebraically, in the starter for example of the present invention, yeast adopts the first-generation to expand numerous female bacterium bacterium liquid, other three kinds of bacterium all adopt the third generation with the numerous bacterium liquid of interior expansion, and strict various raw material bacterium fermentation times of control and temperature, the purpose of doing like this is to make the bacterium in the starter to have more stable proliferation activity, because microorganism many generations expand numerous or expand numerous time oversize after, expand numerous production that obtains microbial inoculum with it for female bacterium again, its activity and character are not difficult to keep efficient stable; And the microorganism that the present invention is to provide in the starter is expanded numerous mistake twice at most, the user can buy a spot of starter of the present invention and ferment voluntarily and expand numerous a large amount of can directly use and compound microbial culture starter that effect guarantees that obtain, and used nutrient solution is with low cost, and the raiser spends the cost on microbial starter culture almost can ignore like this.
In the preferred embodiments of the present invention, preferred fermentation time and the temperature of various raw material bacterium in the compound microbial culture starter is provided, the inventor is by a series of simultaneous test, the optimum blending ratio that obtains various raw material bacterium bacterium liquid that has also drawn under above-mentioned its preferred fermentation time and temperature is a yeast: subtilis: bacillus natto: four kinds of bacterium liquid of milk-acid bacteria 3: 3: 3 by volume: 1, under this ratio, can farthest bring into play the synergy between the various bacterium, what make compound microbial culture starter of the present invention has taken into account the decomposition efficiency height, being used for fodder additives improves the most apparent to animal production, improve the animal resistibility, significantly reduce sickness rate, advantage such as effect is stable.
The present invention also provides the application method of above-mentioned compound microbial culture starter in livestock and poultry cultivation invention, simple and clear, economical, with the supporting use of starter of the present invention, can under the prerequisite that guarantees the cultured product quality, reduce aquaculture cost to the utmost for numerous raisers.
Method when the present invention also further provides the application of compound microbial culture starter of the present invention in the livestock and poultry cultivation each side, but the application of starter of the present invention is not limited to these methods.
Embodiment
Microbial strains and source thereof that the present invention adopts:
The former bacterium of yeast saccharomyces cerevisiae (Saccharomyces cerevisiae CICC31011 Sichuan Food Fermentative Industry Design Academy).
Subtilis (separation of the Bacillus subtilis CICC10036 first fermentation industry Science Institute of Ministry of Light Industry)
Bacillus natto bacterial strain (Bacillus natto CICC20132 Shanghai City Industry Wei Biological Research Institute (B167))
Lactic acid coccus (key lab of the Lactococcus lactis CICC22861 dairy science the Ministry of Education separates (4.0610))
More than four bacterial strains, this experiment all has preservation, can provide to the public from 20 years applyings date, uses as research.
Embodiment 1
(1) culture of strains
1. cultivation method for saccharomycete
The preparation of nutrient solution: yeast extract paste 0.5%, peptone 0.5%, glucose 0.5% is produced in the jar, heats to 90 ℃, is keeping 2 hours more than 90 ℃.Be cooled to then about 33 ℃.Insert the former bacterium of 0.01% yeast saccharomyces cerevisiae (Saccharomycescerevisiae CICC 31011 Sichuan Food Fermentative Industry Design Academy).
Culture condition: under 33 ℃～34 ℃, stir culture 12 hours.
It is stand-by that the yeast liquid of producing is put into 4 ℃ of refrigerations.
2. subtilis cultural method
2.1 culture of strains
2.1.1 the preparation of nutrient solution
The flat-plate solid substratum: extractum carnis 0.5%, yeast extract paste 0.5%, glucose 0.5%, peptone 1%, Nacl 0.5%, 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%, triangular flask is put in the dilution of adding distil water or deionized water, 121 ℃ of autoclavings 20 minutes.Naturally cool to then about 30 ℃.
2.1.2 the enlarged culturing of bacterial classification
Flat-plate solid one-level enlarged culturing: under aseptic condition, getting a fritter freeze-drying lactobacillus (separation of Bacillus subtilis CICC 10,036 first fermentation industry Science Institutes of Ministry of Light Industry) dissolves with 0.5ml sterile purified water or deionized water dilution, then it is uniformly coated on the solid medium, puts into more than 31 ℃～32 ℃ thermostat containers to 48 hour.Under aseptic condition, it is stand-by that the bacterial classification of one-level cultivation is put into sterile chamber with sterile purified water or deionized water flushing.
Liquid secondary enlarged culturing: under aseptic condition, the bacterium liquid that one-level is cultivated inserts in the liquid nutrient medium by 0.5%, and under 31 ℃～32 ℃, 180～200rpm cultivated 33 hours.
2.2 produce 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 less than 80 order particles, puts into and produce jar, heat,, be cooled to then about 31 ℃ keeping 2 hours more than 90 ℃ to 90 ℃.
2.2.2 inoculum size inserts 1% secondary enlarged culturing strain liquid toward producing in the jar.
2.2.3 under 31 ℃～32 ℃ of the culture condition, stir culture 33 hours.
It is stand-by that the bacillus liquid of producing is put into 4 ℃ of refrigerations.
3. the cultural method of bacillus natto
The substratum of bacillus natto bacterial strain (Bacillus natto CICC20132 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. lactic acid coccus enlarged culturing method
4.1 culture of strains
4.1.1 the preparation of nutrient solution
Flat-plate solid substratum: casein peptone 1%, beef extract 1%, yeast extract 0.5%, glucose 0.5%, sodium acetate 0.5%, dihydrogen citrate 0.2%, K2HPO40.2%, MgSO4.7H2O 0.02%, MnSO4.H2O 0.05%, agar 15% gram, 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%, MgSO4.7H2O 0.02%, and MnSO4.H2O 0.05%, PH6.8.Sealable bottle is put in the dilution of adding distil water or deionized water, 121 ℃ of autoclavings 20 minutes.Naturally cool to then about 36 ℃.
4.1.2 the enlarged culturing of bacterial classification
Flat-plate solid one-level enlarged culturing is under aseptic condition, getting a fritter freeze-drying lactobacillus (Lactococcus lactis CICC 22861 key labs of the dairy science the Ministry of Education separate (4.0610)) dissolves with 0.5ml sterile purified water or deionized water dilution, then it is uniformly coated on the solid medium, puts into 36 ℃～37 ℃ anaerobic thermostat containers to 36 hour.Under aseptic condition, it is stand-by that the bacterial classification of one-level cultivation is put into sterile chamber with sterile purified water or deionized water flushing.
Liquid secondary enlarged culturing is under aseptic condition, and the bacterium liquid that one-level is cultivated inserts in the liquid nutrient medium by 1%, and container closure, under 36 ℃～37 ℃, anaerobism was cultivated 36 hours.
4.2 produce method with lactobacillus suspension
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%, MgSO4.7H2O 0.02%, and MnSO4.H2O 0.05%, PH6.8.Put into and produce jar, heat,, be cooled to then about 36 ℃ keeping 2 hours more than 90 ℃ to 90 ℃.
4.2.2 inoculum size inserts 1% secondary enlarged culturing strain liquid toward producing in the jar.
4.2.3 under 36 ℃～37 ℃ of the culture condition, static cultivation 36 hours
It is stand-by that the bacillus liquid of producing is put into 4 ℃ of refrigerations.
Embodiment 2 assembly compound microbial culture starters
Step 1. is mixed the bacterium liquid that makes among the embodiment 1 by following volume ratio:
Subtilis: bacillus natto: yeast: milk-acid bacteria=3: 3: 3: 1 mixed, than adding stablizer, coating-dividing sealing is preserved under the lucifuge condition according to ten thousand/volume mass.
Stablizer is a SODIUM PHOSPHATE, MONOBASIC: Sodium phosphate dibasic is pressed 2: 1 mixed preparing of mass ratio and is got.
Step 2. product detects
Yeast viable count 〉=10
8
Subtilis viable count 〉=10
7
Bacillus natto viable count 〉=10
7
Viable lactic acid bacteria number 〉=10
7
The application of embodiment 3, compound microbial culture starter
Step 1 expands numerous
1. get the container more than 10 kilograms, as water vat, plastic tank, stainless steel cask all can, clean up, do not have any greasy, chemical substance is residual.
2. 1 kilogram of brown sugar is poured in 10 kg of water, be stirred to abundant dissolving and boil, naturally cool to then about 35 ℃, add 1 kilogram of the compound microbial culture starter that embodiment 2 makes, after fully stirring, cultivation naturally.
3. in the culturing process, envrionment temperature remains on 30～35 ℃, stirs once, and just becomes probiotics after 48 hours in per 4～8 hours.
4. after cultured bacterium liquid stirred, available little mineral water bottle was scooped out one bottle and is left standstill observation, expands numerous effect bacterium liquid preferably, the skim white depositions can occur after a few hours at the bottom of bottle.
(2) precaution
1. in the culturing process, the most handy paper of vessel port or gauze cover to be tightened, not uncovered always, to reduce other living contaminants.
2. temperature is low, and incubation time will extend, if envrionment temperature is lower than 22 ℃, bacterial classification stops growing.
3. the microbial inoculum of probiotics stoste or enlarged culturing will be avoided solar radiation in preservation and culturing process, because of the ultraviolet ray in the sunlight can be killed probiotic bacterium.
4. cultured microbial inoculum is preferably in 3～7 days and uses up, and as toing many or too much for use temporarily, should seal cryopreservation, and preservation period surpasses 30 days, as stench occurs and do not use.The user can determine to cultivate quantity according to the breed needs of oneself, the long-time preservation of trying not.
Probiotics stoste can only enlarged culturing once, can not many generations expand numerous, in order to avoid influence result of use.
(3) using method
The microbial inoculum of enlarged culturing can be used for the ecologic breeding of pig, chicken.Before the use, fully shake up.
In the process of using probiotic bacterium, feed and drinking-water do not add any microbiotic, if ill domestic animal will be isolated for treatment separately.If need full group medication, temporarily stop using probiotic bacterium, until drug withdrawal.
1. drink water: the first use, the drinking-water dosage can maintain 0.3～0.5% after 1%, 10 day in preceding 10 days.
2. spice: bacterium liquid adds in the complete diet pellet by 0.5%, mixes thoroughly and feeds, and fed the same day, and mixed the same day.The direct charge level of also available diluent sprays.
3. spraying: by 0.1% dilution, carry out the sterilization of hen house band poultry, can play the effect of depositing dust and environment purification as thimerosal.
4. making fermented feed: according to feed: water: bacterium liquid: brown sugar=100: 60: 1: 1 ratio is mixed all raw materials thoroughly, pack in the sealing bag, place the warm place fermentation of lucifuge 5-7 days, it is salubrious to open bag sight material, nothing is gone mouldy, and has wine flavour to show and ferments successfully.Join in the complete diet pellet by 10% of feeding capacity, mix thoroughly and feed.Fermented feed uses up behind the Kaifeng as early as possible, is no more than two days winter, is no more than 1 day summer.
5. be used for fermenting bed padding: the bacterium liquid after 1 kilogram of expansion is numerous can be made 5 cubical bedding and padding, need not additionally add auxiliary materials such as Semen Maydis powder, wheat bran or brown sugar.Making method is as follows: adopt simple carvel built, spread 20 centimeters rice husk or corn cob meal or roll short corn stalk at the circle end earlier, spray bacterium liquid and water, repave 10 centimeters sawdust or rice husk or Pericarppium arachidis hypogaeae on it, water spray and bacterium liquid, and do agitation as appropriate, make moisture even.Up to being paved with circle body, 80 centimeters are dark by that analogy, 10 centimeters in last one deck shop sawdust, and water spray and bacterium liquid on it, stirring makes moisture distribution even, covers gunnysack or woven bag on it.The following 30 centimeters temperature of bedding and padding are raised to more than 35 ℃ and just can advance swine rearing after 3-7 days.Annotate: expand numerous microbial inoculum and add that the gross weight of the water of spray is 30%～40% of a dunnage weight.
6 above usages can be used separately, also can be used in combination.
The life-time service probiotic bacterium can reach strengthening immunity, suppresses the generation of bacteriosis, and it is extremely naughty to reduce, and improves breeding environment, reduces the animal house stench, and cultures medication by reducing, and reaches and improves meat quality and purpose of safety, thereby improve culture benefit.
Embodiment 4 application examples and effect are relatively
1 materials and methods
Selecting cotton stalk powder, sawdust, rice husk for use is that main raw material carries out proportioning, makes the fermentation bed to raise pig bedding and padding
Microbial inoculum: choose the market and sell preferably 2 kinds of fermentation bed special bacteria agents, be designated microbial inoculum No. 1 (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), the compound microbial culture starter that makes with the embodiment of the invention 2 expand the microbial preparation (No. 3 microbial inoculums) that obtains after numerous through the method for embodiment 3 and compare research.
The usage ratio of three kinds of microbial inoculums and method are with embodiment 3＜using method〉5 described methods.
Experimental animal: greatly enhance, grow 107 of white two-way cross pigs, 68 kilograms of mean body weights, feeding period 30 days
Test design: need 6 swinerys, completely random design, 3 kinds of microbial inoculum * 2 repetitions, totally six colony houses are adopted in the evaluation test of bacterial classification.
Grouping sees the following form 1:
| Numbering | ??1# | ??2# | ??3# | ??4# | ??5# | ??6# |
| The 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 is by hurdle record feed amount every day.Automatic drinking bowl is freely drunk water.Every test pig all will be carried out the overbit mark, and list is only weighed.Feed nutrition level unanimity, immune programme for children is identical.
3 testing indexs
Production performance index: day weight gain, day feed consumption, material anharmonic ratio
The padding bed operation conditions of fermenting:
Temperature measuring: measure the temperature of fermentation bed surface and degree of depth 10cm, 30cm, 50cm weekly respectively, every circle is chosen 5 points; Measure colony house internal and external temperature, humidity simultaneously.
Bedding and padding water content: gather simultaneously with the sample of carbon-nitrogen ratio, adopt the moisture content in National Standard Method " measuring method of moisture in the feed " the detection bedding and padding.
Total plate count: gather simultaneously with the sample of carbon-nitrogen ratio, adopt blood cell plate counting process to measure total viable count pH value
4 data analyses
Adopt the spss10.0 statistical analysis software, the data that test is gathered are carried out statistical treatment.
5 results of study and analysis
5.1 padding bed production performance research is measured
Testing data shows, there is notable difference in the padding bed production performance influence to growing and fattening pigs that different microbial inoculums are made, studies selected 3 kinds of commodity microbial inoculums, and wherein No. 3 microbial inoculums have production performance preferably, its day weight gain and day feed consumption and material anharmonic ratio index all obviously are better than other two kinds of bacterium, see Table 2.The material anharmonic ratio is respectively than No. 1 and No. 2 bacterium reductions by 14.2% and 9.0%.
Table 2 uses the influence of the fermentation bed of different fermentations agent making to the pig production performance
5.2 the Changing Pattern of total viable count in the bedding and padding operational process
Changing Pattern to total viable count in the various bedding and padding in the breeding process has carried out tracking and measuring, and measuring method is a blood cell plate counting.
Sampling method: in each hurdle house according to the principle collected specimens of 5 point samplings, directly get the 0-15 centimetre of abundant mixing in top layer after, according to quartering, get 1 part of pack, be used in reference to target mensuration.
Data analysis shows, and sees Table 3, and total count is along with the prolongation of feeding time presents the trend that increases gradually in the bedding and padding, and on average every weekly gram bedding and padding increase 1.5-2.5 hundred million viable bacterias.
Total viable count changes measured value (hundred million/gram) in time in the bedding and padding that the agent of table 3 different fermentations is made
Data presentation: the Changing Pattern basically identical of the bedding and padding total viable count that different microbial inoculums are made, but the bacterium number there are differences, and microbial inoculum 3 bedding and padding viable bacteria contents are apparently higher than microbial inoculum 2 and 1, and microbial inoculum 2 viable counts are minimum.Illustrate that different microbial inoculums there are differences, microbial inoculum 3 activity are stronger, and reproductivity resistance is all obviously because other two kinds of microbial inoculums.
The huge fluctuation of later stage total viable count is the testing error that causes because of the measuring method difference, non-padding bed practical situation.5.3 padding bed variation of temperature rule in the bedding and padding breeding process
Each degree of depth bedding and padding temperature-averaging value of padding bed each position that table 4 different strain is made (℃)
The padding bed identical change rule of following of different microbial inoculums, 1# and 3# microbial inoculum bed tempertaure measured value basically identical, and No. 2 microbial inoculum bed temperatures are obviously on the low side, the full phase average on the low side 2.7 is spent.The padding bed difference that truly has of different microbial inoculums making is described.Padding bed temperature accumulated temperature reflects the active degree of the breeding of microbial reproduction wherein, and the higher fermentation bed of bed temperature is higher to the glycolysis ability of swine excrement, and this conforms to the measurement result of total count in the bedding and padding.The active degree that microbial inoculum of the present invention is described is higher, and ight soil glycolysis ability is stronger.
5.4 the variation of pH value in the bedding and padding application process
Test has been carried out tracking monitor to the variation of bedding and padding pH value in the bedding and padding breeding process, and sampling weekly takes by weighing 1.0 grams in the sample, place the 100ml beaker, adds the 40ml deionized water.Measure the pH value of solution value with Thermo Orion acidometer behind the dipping 30min.The result is as follows:
The fermentation bed pH that the different microbial inoculums of table 5 are made changes
Acid-basicity in the pH value reflection bedding and padding of bedding and padding, the acid mistake exceeds low breeding and the growth that all is unfavorable for bacterium in the bedding and padding, different bacterial classification combinations may form symbiosis homeostasis preferably in bedding and padding, keep suitable acid-basicity, guarantee self normal growth and breeding.
Data presentation from table 5 get rid of to be measured or sampling error, and the pH of 3 kinds of microbial inoculums does not have notable difference about 60 days, but along with the prolongation of feeding time, No. 2 microbial inoculum pH are apparently higher than No. 3 bacterium and No. 1 bacterium, and the glycolysis ability that reflects No. 2 bacterium is not as No. 3 bacterium.
5.5 the variation of moisture content in the bedding and padding application process
The water content of bedding and padding is determining the success or not of padding bed application to a great extent, and water content is too low, can cause not only that microbial inoculum is active to be reduced, and a large amount of dust can increase pig and suffers from respiratory tract disease.And moisture is too high, can be subjected to severe inhibition by bacterial reproduction again, and the bed surface supporting capacity reduces rapidly, finally causes padding bed routed comprehensively.Therefore the definite suitable water content of hydrolysis and fermentation bed operating is most important for the maintenance of bed surface.
The padding bed change of soil water content (%) of table 6 different strain
Discover, padding bed making initial stage water content about 60%, prolongation along with feeding time, moisture in the bedding and padding constantly evaporates, water content descends gradually, and 4-5 is after week, and water content maintains 40%-50% substantially, the fecaluria moisture of pig has reached in a basic balance with fermentation bed transpiring moisture, and the bed surface situation is better.
There is certain influence in different microbial inoculums to the water content of bedding and padding, and under same stocking density, No. 2 the bacterium group has one to iris out existing moisture rising, the bearing capacity that bedding and padding are described obviously descends, be in the edge of collapse, and No. 1 bacterium and No. 3 bacterium moisture are more suitable, do not have the risk of dead bed.
Conclusion
Show 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 can keep the normal fermentation state of fermenting bed padding, guarantee the efficient glycolysis of fecaluria, reduce the maintenance and the use cost of bedding and padding.This has crucial meaning in production application.
Claims (10)
1. compound microbial culture starter, living microorganism wherein comprises yeast, subtilis (Bacillus subtilis), bacillus natto (Bacillus natto) and milk-acid bacteria, described yeast is yeast saccharomyces cerevisiae (Saccharomyces cerevisiae), and described milk-acid bacteria is lactic acid coccus (Lactococcus lactis).
2. compound microbial culture starter according to claim 1, described yeast are 0.01% to insert the former bacterium of yeast by volume in the liquid medium within, cultivate down for 33 ℃～34 ℃ and make bacterium liquid in 10～15 hours;
Described subtilis be in the liquid medium within by the Bacillus subtilis strain liquid of 1% inoculation secondary enlarged culturing, under 31 ℃～32 ℃, cultivate the bacterium liquid of 30～35 hours gained;
Described bacillus natto be in the liquid medium within by the bacillus natto strain liquid of 1% inoculation secondary enlarged culturing, under 31 ℃～32 ℃, cultivate the bacterium liquid of 30～35 hours gained;
Described milk-acid bacteria be in the liquid medium within by the 1% lactic acid bacteria culturers liquid that inserts the secondary enlarged culturing, under 36 ℃～37 ℃, the bacterium liquid of 30～40 hours gained of static cultivation.
3. compound microbial culture starter according to claim 2, described yeast, subtilis, bacillus natto, four kinds of bacterium liquid of milk-acid bacteria 3: 3: 3 by volume: 1 mixes.
4. claim 1 or the 2 or 3 described compound microbial culture starters application in livestock and poultry cultivation is produced is characterized in that earlier described compound microbial culture starter being expanded numerous numerous microbial inoculum that obtains expanding, and the numerous step of described expansion is as follows:
(1) brown sugar was mixed than 1: 10 by weight with water and be stirred to abundant dissolving, be cooled to 34～36 ℃;
(2) add 1 part of described compound microecological starter, fully stir, place 30～35 ℃ of environment, stirred once, cultivated 45～50 hours in per 4～8 hours.
5. application according to claim 4 refers to be used for livestock and fowl drinking water, it is characterized in that using preceding 10 days, add by 1% in drinking-water and expand numerous microbial inoculum, but add-on is 0.3～0.5% after the 10th day.
6. application according to claim 4 refers to that the numerous microbial inoculum of described expansion adds in the complete diet pellet by 0.5%, and described adding refers to mix thoroughly or spray on the complete diet pellet surface with complete diet pellet.
7. application according to claim 4 refers to the numerous microbial inoculum of described expansion by 0.1% dilution, sprays in the breeding environment of livestock and poultry to carry out disinfection.
8. application according to claim 4 refers to according to feed: water: expand numerous microbial inoculum: brown sugar=100: 60: 1; 1 volume ratio is mixed thoroughly, and lucifuge fermentation obtained fermented feed in 5-7 days in the sealing bag of packing into, by 10% of feeding capacity described fermented feed is joined in the complete diet pellet, mixes the livestock and poultry of feeding thoroughly.
9. application according to claim 4 refers to make fermenting bed padding, the mass volume ratio 1kg of numerous microbial inoculum of described expansion and bedding and padding raw material: 5M
3, described bedding and padding raw material and microbial inoculum mixed fermentation at normal temperature are used for its feeding after bedding and padding stockpile internal temperature is raised to more than 35 ℃, and described microbial inoculum is mixed into for expanding numerous microbial inoculum and water, and described microbial inoculum accounts for 30～40% of dunnage weight.
10. application according to claim 9, described mixed fermentation refers to spread at the circle end earlier that the rice husk, corn cob meal of 20 cm thicks are or/and roll short corn stalk, spray microbial inoculum, the sawdust, rice husk that repave 10 centimeters be or/and Pericarppium arachidis hypogaeae, the spray microbial inoculum, with this shop of circulating to 80 cm thicks, spread the sawdust of 10 cm thicks at last, spray microbial inoculum in surface covers gunnysack or woven bag then, treats that the following 30 centimeters temperature of bedding and padding are raised to more than 35 ℃ to get final product.
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