CN1806659A - Method for preparing microbial feed addictive by multiple bacteria strain mixed culture - Google Patents
Method for preparing microbial feed addictive by multiple bacteria strain mixed culture Download PDFInfo
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- CN1806659A CN1806659A CNA200610031230XA CN200610031230A CN1806659A CN 1806659 A CN1806659 A CN 1806659A CN A200610031230X A CNA200610031230X A CN A200610031230XA CN 200610031230 A CN200610031230 A CN 200610031230A CN 1806659 A CN1806659 A CN 1806659A
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Abstract
The invention relates to a method of producing microbe feed stuff addition agent by multi-bacteria, among which, the bacteria is: plant lactobacillus, Lactobacillus acidophilus, fibroin-producing candida, Saccharomyces cerevisiae; make fibroin-producing candida and Saccharomyces cerevisiae into saccharomycete mixing seed liquid; make plant lactobacillus and Lactobacillus acidophilus into lactobacillus mixing seed liquid; inoculate said saccharomycete mixing seed liquid and lactobacillus mixing seed liquid to saccharomycete and lactic acid bacteria mixing seed culture medium to get saccharomycete and lactic acid bacteria mixing seed liquid; inoculate saccharomycete and lactic acid bacteria mixing seed liquid at the quantity of 0.2-0.3% to the completed product produced through grading mixing fermentation in mixing fermentation culture medium; The mixing fermentation culture of saccharomycete and lactic acid bacteria is divided into two stages, the earlier stage is aerobic culture, and the later stage is anaerobic culture. The invention is characterized on low cost of production and low energy consumption; the bacteria could cooperate with each other well which enhances the phylactic power of animals and the availability ratio of feed stuff; the method relates to the multi-bacterial strain mixing culture of microbe feed stuff addition agent which accelerates the health growth of animals
Description
Technical field
The present invention relates to a kind of production method of probiotics, particularly adopt the method for multi-strain bacteria kind Mixed culture probiotic feed additive.
Background technology
Bread is the staff of life.Food-safety problem is the significant problem that is related to people ' s health and national economy, and particularly after China's WTO, the agricultural product security problem has become the key factor that influences the agricultural products in China industrial competitiveness.Pork is the main animal food that people absorb, and safe feed is the basis of animal food safety.Develop safe in utilizationly, the feed additive of favourable pig growth is the key of development " nuisanceless pig raising technology ", and the microorganism fodder preparation is the good method of the first-selection of relatively generally acknowledging at present.Along with extensively carrying out of microorganism application technology, in the application facet of active bacteria formulation, people wish that a kind of product can exercise multiple function, solve many-sided problem, and many products all are made up of multiple microorganism.At present, on the production form of this series products, the general employing separately cultivated single culture, and the culture with multiple bacterium mixes then, forms many bacterial classifications multifunctional product.This mode of production requires every kind of bacterium to use the one time fermentation jar, requires investment of production equipment bigger, and energy consumption is higher, has improved production cost greatly.Present for the bacterial strain screening with synergy relation and combination or a random process, lack effective theoretical direction, and can not coordinate relation between bacterium effectively for the Mixed culture system of having used, make it reach best ecological level, the performance ceiling effect.This has seriously hindered development and application that Mixed Microbes is cultivated.
Summary of the invention
Purpose of the present invention aims to provide and a kind ofly can reduce production costs well, energy consumption, can act synergistically well mutually between bacterial classification, improve the immunity of animal, improve efficiency of feed utilization, promote the method for the multi-strain bacteria kind Mixed culture production additive for microbe feedstuff of animal health growth.
The objective of the invention is to realize by following manner:
Employed bacterial classification is: Lactobacillus plantarum (Lactobacillus plantarum), lactobacillus acidophilus (Lactobacillus acidophilus), candida utili (Candida tail.s), saccharomyces cerevisiae (Saccharomyces cerevisiae); Its technology is: saccharomyces cerevisiae and candida utili are made saccharomycete seed mixture liquid; Lactobacillus plantarum, lactobacillus acidophilus are made lactic acid bacteria seed mixture liquid; Above-mentioned resulting saccharomycete seed mixture liquid, lactobacillus acidophilus seed mixture liquid are inoculated in saccharomycete and the lactic acid bacteria seed mixture culture medium, obtain saccharomycete and lactic acid bacteria seed mixture liquid; Saccharomycete and lactic acid bacteria seed mixture liquid is inoculated in the mixed fermentive culture medium by the inoculum concentration of 0.2-0.3% obtains finished product after the segmentation mixed culture fermentation; Described mixed fermentive culture medium is: 1-3% brown sugar, 1.5% glucose, 0.1-0.3% peptone, 0.5-1.5% dregs of beans, 0.1-0.6% radish Juice, 0.1% K
2HPO
4, 0.1% MgSO
4.7H
2O, water 1000mL, pH 6.4-6.6; The segmentation mixing fermentation culture of described saccharomycete and lactic acid bacteria is carried out in two stages, aerobic cultivation: cultivation temperature 28-30 ℃, throughput 300L/min whenever led to 1h and stopped 10min, speed of agitator 40-50r/min early stage, every 3h 30min stops 30min, cultivates 20-30h; Later stage anaerobic fermentation: cultivation temperature 32-34 ℃, throughput 0L/min, speed of agitator 40-50r/min, every 30min stops 30min, cultivates 20-30h.
Also add the 0.0-0.3% vegetable oil in the described mixed fermentive culture medium.
Best results when mixed fermentive culture medium is following component: 2% brown sugar, 1.5% glucose, 0.1% peptone, 1% dregs of beans, 0.5% radish Juice, 0.1% K
2HPO
4, 0.1% MgSO
4.7H
2O, 0.15% vegetable oil, water 1000mL transfer pH 6.4-6.6.
Saccharomycete seed mixture liquid, lactobacillus acidophilus seed mixture liquid are inoculated in saccharomycete and the lactic acid bacteria seed mixture culture medium by the inoculum concentration of 1-2%, 0.5-1.5% respectively.
Saccharomycete seed mixture liquid, lactobacillus acidophilus seed mixture liquid are inoculated in saccharomycete and the lactic acid bacteria seed mixture culture medium by 1.5%, 1% inoculum concentration respectively; Saccharomycete and lactic acid bacteria seed mixture liquid is inoculated in by 0.2% inoculum concentration carries out mixed culture fermentation, best results in the mixed fermentive culture medium.
The present invention selects with bacterial classification by repeatedly studying and test for producing, and, Lactobacillus plantarum (Lactobacillus plantarum), lactobacillus acidophilus (Lactobacillus acidophilus), candida utili (Candida tails), saccharomyces cerevisiae (Saccharomyces cerevisiae) carried out research, test by being produced with characteristic and the performance of bacterium for 4 kinds, test, the inventor finds that effective combination of above-mentioned 4 kinds of bacterial classifications has effect well for product of the present invention.And bacterial classification used in the present invention is the feed grade probio bacterial classification that the Ministry of Agriculture allows use.The present invention utilizes saccharomycete to be facultative aerobe, lactic acid bacteria is microaerophilic characteristics, advanced person's gas cultivation of acting charitably, make saccharomycete produce a large amount of thalline and metabolite, this condition of culture does not damage the survival of lactic acid bacteria again simultaneously, after cultivating early stage, stopping ventilation cultivating, make saccharomycete can carry out alcoholic fermentation, produce a certain amount of alcohol and carbon dioxide, for the generation of ester perfumery matter and little aerobic growth of lactic acid bacteria provide condition, lactic acid bacteria can utilize saccharomycetic metabolite to grow simultaneously, further reduces the pH value, for the pollution that reduces the assorted bacterium of product creates conditions, and reduced energy consumption.
The present invention utilizes saccharomycetic medium component simple, metabolite is nutritious, and the medium component complexity of lactic acid bacteria, and price is more expensive, saccharomycetic metabolite or dead cell autolysate can provide the nutritional requirement of lactobacter growth, and research optimization has obtained Lactobacillus plantarum (Lactobacillus plantarum), lactobacillus acidophilus (Lactobacillus acidophilus), candida utili (Candida tails), the culture medium prescription of 4 kinds of bacterium Mixed culture of saccharomyces cerevisiae (Saccharomyces cerevisiae) (i.e. 2% brown sugar, 1.5% glucose, 0.1% peptone, 1% dregs of beans, 0.5% radish Juice, 0.1% K
2HPO
4, 0.1%MgSO
4.7H
2O, 0.15% vegetable oil, water 1000mL, pH 6.4-6.6.Greatly reduce the peptone content in the general lactic acid bacteria culture medium, saved dusty yeast (cream) composition, and adopted low price, abundant bean cake powder to do nitrogenous source, and adopt the red pool to replace part sucrose or glucose to do carbon source, reduce about 1510.9 yuan/ton of raw materials for production costs.
The advantage of maximum of the present invention is, the present invention proposes Lactobacillus plantarum (Lactobacillusplantarum), lactobacillus acidophilus (Lactobacillus acidophilus), candida utili (Candida tails), saccharomyces cerevisiae (Saccharomyces cerevisiae) are carried out Mixed culture, the stepwise fermentation technology has creatively been proposed, mixed culture fermentation main aerobic culture yeasts bacterium in early stage of the present invention obtains thalline and metabolite, and main anaerobism of later stage is cultivated lactic acid bacteria and obtained thalline and make saccharomycetes to make fermentation produce wine flavour.And worked out well at four kinds of bacterium mixed fermentive culture mediums of the present invention simultaneously, four kinds of bacterium mixing fermentation culture conditions, these all do not have the pertinent literature report.And the crux that product of the present invention can successfully prepare also is determining of above-mentioned condition.
Product of the present invention is used for feeding of pig and chicken to be had growth promotion, improve feedstuff-meat ratio, improves immunity, reduces functions such as the environmental pollution and the generation that wards off disease animal; Can improve the vigor of boar sperm, reduce rate of teratosperm; There are not hormone and antibiotic residue in the meat product, the resistance to the action of a drug and medicament residue have been avoided, of the present inventionly applying the competitiveness in the international market that can improve China's pork product, guarantee the quality and the safe and sanitary of pork product, is the new generation product after antibiotic.Thereby the present invention be a kind of effectively utilize 4 kinds of bacterium well mixed culture fermentation produce the production method of this excellent product of additive for microbe feedstuff, the present invention can reduce production costs greatly, promotes applying of this series products.
Product of the present invention in the ChangSha, Hunan Province, part districts and cities such as Liuyang, Ningxiang, Lanshan County, Zhuzhou and large-scale pig farm promote, and delivers 260,000 bull pigs for sale, the raiser on average increases income more than 50 yuan/head, direct economic benefit reaches more than 1,500 ten thousand yuan.
The specific embodiment
Embodiment 1
For example the present invention is done more detailed description below:
Bacterial classification: candida utili [ACC20060], saccharomyces cerevisiae [ACC20065], Lactobacillus plantarum [ACC10644], lactobacillus acidophilus [ACC10637] (being the purchase of Chinese agriculture microorganism fungus kind preservation center obtains).
Respectively above-mentioned four kinds of bacterial classifications are carried out actication of culture, candida utili, saccharomyces cerevisiae are inoculated in respectively on the PDA slant medium, 20-30 ℃, cultivate 30-40h; After the activation, cultivate respectively and produce candida utili, saccharomyces cerevisiae liquid seeds continuously; And Lactobacillus plantarum, lactobacillus acidophilus are inoculated in the MRS slant medium respectively, 30-40 ℃, cultivate 30-40h; After the activation, cultivate respectively and produce Lactobacillus plantarum, lactobacillus acidophilus liquid seeds continuously.
With saccharomyces cerevisiae and candida utili combined inoculation in the YPD seed culture medium, under 26-30 ℃, 160-180r/min, cultivate 20-30h, obtain the saccharomycete seed mixture, and then be inoculated in the saccharomycete seed mixture culture medium in 26-30 ℃ 160-180r/min, cultivate 20-30h, get saccharomycete seed mixture liquid.Being prepared as of saccharomycete seed mixture culture medium wherein: peptone 10.0g, glucose 20.0g, yeast extract 5.0g, distilled water 1000mL, adjust pH are 6.8～7.0, sterilization treatment is stand-by.The inventor finds also in the saccharomycete seed mixture liquid that it is 1 that saccharomyces cerevisiae is counted ratio with the bacterium of candida utili: during 1-3, can optimize product of the present invention.
Lactobacillus plantarum, lactobacillus acidophilus combined inoculation in the MRS seed culture medium, under 30-34 ℃, are cultivated 20-30h, obtain the lactic acid bacteria seed mixture, and then be inoculated in the lactic acid bacteria seed mixture culture medium under 30-34 ℃, cultivate 20-30h, lactic acid bacteria seed mixture liquid.Wherein lactic acid bacteria seed mixture medium preparation is: peptone 10g, beef extract 10g, yeast extract 5g, dipotassium hydrogen phosphate 2g, dibasic ammonium citrate 2g, sodium acetate 5g, glucose 20g, Tween 80 1mL, epsom salt 0.58g, four water manganese sulfate 0.25g, water 1000mL, adjust pH 6.2-6.4, sterilization treatment, stand-by.The inventor finds that also it is can optimize product of the present invention at 1: 12 o'clock that the bacterium of Lactobacillus plantarum, lactobacillus acidophilus in the lactic acid bacteria seed mixture liquid is counted ratio.
Above-mentioned resulting saccharomycete seed mixture liquid, lactobacillus acidophilus seed mixture liquid are inoculated in saccharomycete and the lactic acid bacteria seed mixture culture medium by the inoculum concentration of 1-2%, 0.5-1.5% respectively, under 30-34 ℃, 160-180r/min, cultivate 30-40h, obtain saccharomycete and lactic acid bacteria seed mixture liquid.Described saccharomycete and lactic acid bacteria seed mixture medium preparation are: 2% brown sugar, 1.5% glucose, 0.5% peptone, 0.5% radish Juice, 0.1% K
2HPO
4, 0.1% MgSO
4.7H
2O, 0.15% vegetable oil, water 1000mL, adjust pH 6.4-6.6; Sterilization treatment, stand-by.
Saccharomycete and lactic acid bacteria seed mixture liquid is inoculated in by the inoculum concentration of 0.2-0.3% carries out mixed culture fermentation in the mixed fermentive culture medium, mixed fermentive culture medium is by the preparation of following component: 1-3% brown sugar, 1.5% glucose, 0.1-0.3% peptone, 0.5-1.5% dregs of beans, 0.1-0.6% radish Juice, 0.1% K
2HPO
4, 0.1%MgSO
4.7H
2O, 0.0-0.3% vegetable oil, water 1000mL, pH6.4-6.6; Sterilization treatment, stand-by.
The segmentation mixing fermentation culture condition of saccharomycete and lactic acid bacteria is: early stage aerobic cultivation: cultivation temperature 28-30 ℃, throughput 300L/min, whenever logical 1h stops 10min, speed of agitator 40-50r/min, every 3h 30min stops 30min, cultivates 20-30h; Later stage anaerobic fermentation: cultivation temperature 32-34 ℃, throughput 0L/min, speed of agitator 40-50r/min, every 30min stops 30min, cultivates 20-30h.
Last multiparity product quality detects, and can, packing are checked and accepted.
With the prepared product of the present invention, in feed, to add product and directly feed by 0.3% of daily ration, its effect is obvious, and is simple, is worthy to be popularized.But feed after adding 0.3% microorganism fodder preparation fermentation feed, also may can play feeding effect preferably, save cost.
The product of the application of the invention proves that the resulting complex microorganism preparations of the present invention makes the pig colour of skin ruddy, and growth potential is good, and the physique of pig strengthens; Daily gain improves 15.4%; Efficiency of feed utilization improves 11.7%; Coliform quantity in the ight soil reduces, and pernicious gases such as ammonia reduce; The excrement apparent digestibility significant difference of protein and fat; Total phosphorus content reduces in the ight soil.
The product the key technical indexes that the present invention produces:
Living bacteria count: 〉=2.5 * 10
8CFU/mL
Wherein: lactic acid bacteria 〉=2.0 * 10
8CFU/mL
Saccharomycete 〉=0.5 * 10
8CFU/mL
Coliform group count :≤3/100mL.
Salmonella: must not detect.
PH value: 3.0-5.0.
Shelf-life: must not be lower than 6 months (〉=6 months).
Embodiment 2
Saccharomycete seed mixture liquid, lactobacillus acidophilus seed mixture liquid are inoculated in saccharomycete and the lactic acid bacteria seed mixture culture medium by 1.5%, 1% inoculum concentration respectively; Saccharomycete and lactic acid bacteria seed mixture liquid is inoculated in by 0.2% inoculum concentration carries out mixed culture fermentation in the mixed fermentive culture medium.
Mixed fermentive culture medium can reach best by the preparation of following component: 2% brown sugar, 1.5% glucose, 0.1% peptone, 1% dregs of beans, 0.5% radish Juice, 0.1% K
2HPO
4, 0.1% MgSO
4.7H
2O, 0.15% vegetable oil, water 1000mL transfer pH6.4-6.6; Sterilization treatment, stand-by.Other step is with embodiment 1.
Claims (8)
1, the method for preparing microbial feed addictive by multiple bacteria strain mixed culture, employed bacterial classification is: Lactobacillus plantarum, lactobacillus acidophilus, candida utili, saccharomyces cerevisiae; Its technology is: saccharomyces cerevisiae and candida utili are made saccharomycete seed mixture liquid; Lactobacillus plantarum, lactobacillus acidophilus are made lactic acid bacteria seed mixture liquid; Above-mentioned resulting saccharomycete seed mixture liquid, lactobacillus acidophilus seed mixture liquid are inoculated in saccharomycete and the lactic acid bacteria seed mixture culture medium, obtain saccharomycete and lactic acid bacteria seed mixture liquid; Saccharomycete and lactic acid bacteria seed mixture liquid is inoculated in the mixed fermentive culture medium by the inoculum concentration of 0.2-0.3% obtains finished product after the segmentation mixed culture fermentation;
Described mixed fermentive culture medium is: 1-3% brown sugar, 1.5% glucose, 0.1-0.3% peptone, 0.5-1.5% dregs of beans, 0.1-0.6% radish Juice, 0.1%K
2HPO
4, 0.1%MgSO
4.7H
2O, water 1000mL, pH 6.4-6.6;
Described saccharomycete and lactic acid bacteria segmentation mixing fermentation culture are then carried out in two stages, aerobic cultivation: cultivation temperature 28-30 ℃, throughput 300L/min whenever led to 1h and stopped 10min, speed of agitator 40-50r/min early stage, every 3h 30min stops 30min, cultivates 20-30h; Later stage anaerobic fermentation: cultivation temperature 32-34 ℃, throughput 0L/min, speed of agitator 40-50r/min, every 30min stops 30min, cultivates 20-30h.
2, the method for preparing microbial feed addictive by multiple bacteria strain mixed culture according to claim 1 adds the 0.0-0.3% vegetable oil in the mixed fermentive culture medium.
3, the method for preparing microbial feed addictive by multiple bacteria strain mixed culture according to claim 1 is inoculated in saccharomycete and lactic acid bacteria seed mixture liquid and carries out mixed culture fermentation in the mixed fermentive culture medium by 0.2% inoculum concentration.
4, the method for preparing microbial feed addictive by multiple bacteria strain mixed culture according to claim 2, described mixed fermentive culture medium is: 2% brown sugar, 1.5% glucose, 0.1% peptone, 1% dregs of beans, 0.5% radish Juice, 0.1%K
2HPO
4, 0.1%MgSO
4.7H
2O, 0.15% vegetable oil, water 1000mL transfer pH 6.4-6.6.
5, the method for preparing microbial feed addictive by multiple bacteria strain mixed culture according to claim 1, saccharomycete seed mixture liquid, lactobacillus acidophilus seed mixture liquid are inoculated in saccharomycete and the lactic acid bacteria seed mixture culture medium by the inoculum concentration of 1-2%, 0.5-1.5% respectively.
6, the method for preparing microbial feed addictive by multiple bacteria strain mixed culture according to claim 5 is inoculated in saccharomycete seed mixture liquid, lactobacillus acidophilus seed mixture liquid in saccharomycete and the lactic acid bacteria seed mixture culture medium by 1.5%, 1% inoculum concentration respectively.
7, the method for preparing microbial feed addictive by multiple bacteria strain mixed culture according to claim 1, to count ratio with the bacterium of candida utili be 1 to saccharomyces cerevisiae in the saccharomycete seed mixture liquid: 1-3.
8, the method for preparing microbial feed addictive by multiple bacteria strain mixed culture according to claim 1, in the lactic acid bacteria seed mixture liquid bacterium of Lactobacillus plantarum, lactobacillus acidophilus to count ratio be 1: 1-2.
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| CN101946853A (en) * | 2010-08-25 | 2011-01-19 | 天津农学院 | Method for preparing silage feed by fermenting straw through composite microbes |
| CN101703169B (en) * | 2009-11-02 | 2011-12-07 | 许民强 | Method for preparing live lactobacillus planterum preparation and application of live lactobacillus planterum preparation in preventing and treating piglet diarrhoea |
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| CN1135835A (en) * | 1995-05-12 | 1996-11-20 | 洪再发 | Method for breeding chickens |
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| CN109593666A (en) * | 2017-09-30 | 2019-04-09 | 河南牧动物药业有限公司 | A kind of compound micro-ecological preparation and its preparation method and application |
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