CN103211085A - Efficient feed compound probiotics preparation and preparation method thereof - Google Patents
Efficient feed compound probiotics preparation and preparation method thereof Download PDFInfo
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- CN103211085A CN103211085A CN 201310141668 CN201310141668A CN103211085A CN 103211085 A CN103211085 A CN 103211085A CN 201310141668 CN201310141668 CN 201310141668 CN 201310141668 A CN201310141668 A CN 201310141668A CN 103211085 A CN103211085 A CN 103211085A
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Abstract
The invention discloses an efficient feed compound probiotics preparation and a preparation method of the probiotics preparation, belonging to the field of biotechnology. The compound probiotics preparation is prepared by mixing the respectively fermented bacillus licheniformis, streptococcus thermophilus, and saccharomyces cerevisiae hansen according to a ratio, and the number of effective microorganisms reaches 8-120*108CFU/mL. The compound probiotics preparation is scientific and reasonable in strain compatibility, can kill or inhibit pathogenic bacteria in an animal body, promotes feed digestion and absorption, and can greatly reduce the use of antibiotic in the feed.
Description
Technical field
The present invention relates to a kind of feed addictive, relate in particular to a kind of efficient compound probiotic agent of forming by complex microorganism.
Background technology
In modern animal husbandry, add antibiotic in the feed and start from the fifties, to disease preventing and treating, promote growth, save feed, improve the culturing economic benefit and play an important role.But, along with the feed midium or long term is added antibiotic and chemical synthetic drug in large quantities, pathogenic microorganism generation drug resistance in the animal body has appearred in breeding production, and effect reduces, and production cost strengthens; Produce pharmacological dependence in the animal body, the immunity function of self suppresses, and immunity and premunition reduce; Beneficial bacterium is suppressed in the animal body, and flora imbalance forms autogenous infection; Animal product and excreta medicament residue jeopardize health; The diffusion of drug-fast bacteria causes significant problems such as public safety in the animal body, needs to be resolved hurrily.
Therefore, from the eighties beginning, relevant probio is in the application study fast development of aquaculture.At present, probio research both at home and abroad relates to a kind surplus the row beneficial bacterium 80." the feed addictive kind catalogue (2006) " of China's promulgation granted totally 16 of use beneficial bacteriums.The bacillus preparation that promote the domestic market has cereobiogen, whole intestines life, antibacterial life, newborn Kang Sheng etc.; Lactobacillus preparation is based on genus lactubacillus and Bifidobacterium.Probio is as a kind of bioactive additive of new type natural, and is nontoxic, the no resistance to the action of a drug, and noresidue, and have enhancing immunity, and promote growth, improve the effect of efficiency of feed utilization, substitute antibiotic and use in the breeding production.
At present, probiotics preparation is applied to breeding production, but remains aspect technology of preparing in problems: (1) product standard disunity, and manufacturing enterprise sets up standard voluntarily, lacks scientific basis, and probio too high levels or low excessively influences result of use; (2) produce the bacterial classification source and fail to understand that most of enterprises produce and mutually transfer with bacterial classification, do not pass through isolation and selection, spawn degeneration is serious, and physiologically active reduces; (3) the bacterial classification compatibility lacks scientific basis, single bacterial strain, and two bacterial strains, even the random proportioning of many bacterial strains can not illustrate the reasonability of compatibility between bacterial strain not reach the bacterial strain merit that complementary by reasonable compatibility, concertedness is poor; (4) production technology confusion can not be cultivated by the different physiological property differences of bacterial classification, and many manufacturing enterprises adopt many bacterial classifications Mixed culture, and without fermentation tank, very the person adopts and heaps fermentation, causes the probio growth few, and assorted flora is given birth to.
Summary of the invention
In order to solve the not problem of science, synergy difference of compatibility that above probiotics preparation exists, the invention provides the good efficient composite probiotics preparations of scientific and reasonable, the mutual synergistic action effect of a kind of compatibility.
Another object of the present invention provides the preparation method of described efficient composite probiotics preparations.
Compound probiotic agent of the present invention be by bacillus licheniformis (
Bacillus licheniformis), Streptomyces thermophilus (
Streptococcus thermophilus), S. cervisiae (
Saccharomyces cerevisiae Hansen) mix in proportion after the fermented and cultured respectively.
Described bacillus licheniformis agent preparation method is as follows:
(1) bacillus licheniformis preparation
The bacillus licheniformis agent preparation method is as follows:
On the sterilized liquid culture medium, insert the bacillus licheniformis of activation by 2% inoculum concentration, 37 ℃ of aerobic fementations 24 hours can get the bacillus licheniformis preparation;
The mass volume ratio of described fluid nutrient medium nutritional labeling is: 2% starch, 1% soya-bean cake hydrolyzate, 2% corn steep liquor, 1% wheat bran, dipotassium hydrogen phosphate 0.25%, 0.1%, 121 ℃ of sterilization of potassium dihydrogen phosphate 30 minutes;
Described bacillus licheniformis, aerobic growth is cultivated 30 ~ 31 ℃ of thermophilics; Can secrete bacitracin, have broad spectrum antibiotic activity, can kill pathogens such as Escherichia coli, salmonella in the animal intestinal; Can extracellular proteinase, lipase, amylase etc., promote the decomposition of macromolecular substances such as protein in the feed to absorb; Can be heat-resisting, acidproof, under the feed process neutralizes the environment of hydrochloric acid in gastric juice, bile salt in animal body, can keep higher physiological activities, cultivate easily.
(2) Streptomyces thermophilus preparation
The preparation method is as follows in the Streptomyces thermophilus agent:
In the fluid nutrient medium of sterilization, insert Streptomyces thermophilus with mass percent 5% inoculum concentration, 37 ℃ of static cultivations 24 hours can get the Streptomyces thermophilus preparation;
Described fluid nutrient medium nutriment volume ratio is: 40% Fructus Hordei Germinatus hydrolyzate, and 10% soybean hydrolyzate, 10% inorganic salt solution II was sterilized 30 minutes for 121 ℃;
Described inorganic salt solution II is meant and contains sodium acetate 5g, ammonium citrate 2g, dipotassium hydrogen phosphate 2g, magnesium sulfate 0.58g, manganese sulfate 0.25g in the 1000ml solution;
Described Streptomyces thermophilus, anaerobism or facultative aerobic, 40 ~ 42 ℃ of growth temperatures, pH value 5.5 ~ 6.2; Utilize the breeding of nourishing and growing in the enteron aisle in animal body,, suppress the pathogen growth and breeding with pathogen competition nutriment; Can secrete nisalpin, suppress and kill pathogen; Metabolism produces lactic acid, and acetate promotes vitamin D in the feed, and the absorption of calcium, iron ion has additional nutrients.
(3) saccharomyces cerevisiae bacteria preparation
The S. cervisiae bacterial preparation process is as follows:
On the solid medium of sterilization, insert the S. cervisiae of activation with 10% inoculum concentration, 30 ~ 32 ℃ of solid fermentations 36 ~ 48 hours, middle stirring 2 ~ 3 times can get the saccharomycete preparation;
Described solid medium nutritional labeling percentage is:: 40% pomace, 10% wheat bran, 40% malt root, 10% inorganic salt solution I, an amount of water mixes, and sterilizes 30 minutes for 121 ℃;
Described inorganic salt solution I is: contain ammonium sulfate 40g, dipotassium hydrogen phosphate 0.2g, potassium dihydrogen sulfate 0.8g, calcium chloride 0.02g, magnesium sulfate 0.08g in the 1000ml solution;
Described S. cervisiae bacterium protein content height can tolerate hydrochloric acid in gastric juice and bile salt in the animal intestinal and survives; In animal intestinal, can secrete the sanguinariness factor, kill the pathogen in the enteron aisle.
(4) compound probiotic agent preparation
Compound probiotic agent preparation method is: 30 ~ 50 parts in bacillus licheniformis preparation, and 30 ~ 50 parts in Streptomyces thermophilus preparation, the saccharomyces cerevisiae bacteria preparation mixes the liquid-type microbial inoculum for 10 ~ 40 parts; With wheat bran or rice husk absorption, air-dry or 40 ~ 50 ℃ of dryings can get probio solid type microbial inoculum.
Effective microbe content is 8 ~ 120 * 10 in the described compound probiotic agent
9CFU/mL or 6 ~ 150 * 10
9CFU/g
Described compound probiotic agent can 0.1 ~ 0.5% dosage add in animal and fowl fodder or the drinking water and use.
The invention has the advantages that:
(1) the bacterial classification physiologically active height of separation screening, compatibility is scientific and reasonable between bacterial strain, the complementation by reasonable compatibility of bacterial strain merit, concertedness is good, make beneficial microbe colony balance in the animal body, can suppress and kill pathogen, the antibiotic that adds in the substitute feed gradually in the animal body;
(2) bacitracin, nisin and the sanguinariness factor of probio secretion in animal body are the peptide class of gene code, but not the secondary pollutant that mushroom is cultivated, and are decomposed into amino acid etc. voluntarily by metabolism in animal body and are absorbed nontoxic, noresidue;
(3) according to the physilogical characteristics of each bacterial strain, improve cultural method, in the incubation particularly bacillus licheniformis be not easy to infect bacteriophage, be suitable for suitability for industrialized production;
(4) promote animal digestion to absorb, alleviate the ight soil stink, improve feeding environment and ecological environment.
The specific embodiment
For a better understanding of the present invention, further specify below in conjunction with specific embodiment.
Embodiment 1: efficient compound probiotic agent-liquid-type
(1) preparation of bacillus licheniformis agent:
Fluid nutrient medium (W/V): 2% starch, 1% soya-bean cake hydrolyzate, 2% corn steep liquor, 1% wheat bran, dipotassium hydrogen phosphate 0.25%, 0.1%, 121 ℃ of sterilization of potassium dihydrogen phosphate 30 minutes.
On sterilising medium, 2% inoculum concentration inserts the bacillus licheniformis of activation, and 37 ℃ of aerobic fementations 24 hours can get the bacillus licheniformis preparation.
(2) preparation of Streptomyces thermophilus agent:
Fluid nutrient medium (W/V): 40% Fructus Hordei Germinatus hydrolyzate, 10% soybean hydrolyzate, 10% inorganic salt solution II (containing sodium acetate 5g, ammonium citrate 2g, dipotassium hydrogen phosphate 2g, magnesium sulfate 0.58g, manganese sulfate 0.25g in the 1000ml solution) was sterilized 30 minutes for 121 ℃.
In the culture medium of sterilization, with the 5%(percentage by weight) inoculum concentration inserts Streptomyces thermophilus, and 37 ℃ of static cultivations 24 hours can get the Streptomyces thermophilus preparation.
(3) preparation of saccharomyces cerevisiae bacteria agent:
Solid medium (W/W): 40% pomace, 10% wheat bran, 40% malt root, 10% inorganic salt solution I (containing ammonium sulfate 40g, dipotassium hydrogen phosphate 0.2g, potassium dihydrogen sulfate 0.8g, calcium chloride 0.02g, magnesium sulfate 0.08g in the 1000ml solution), an amount of water, mix, sterilized 30 minutes for 121 ℃.
On the culture medium of sterilization, insert the S. cervisiae of activation with 10% inoculum concentration, 30 ~ 32 ℃ of solid fermentations 36 ~ 48 hours, middle stirring 2 ~ 3 times can get the saccharomycete preparation.
With above-mentioned cultured active bacteria formulation, mix according to the ratio of 40 parts of bacillus licheniformis, 40 parts of Streptomyces thermophilus, 20 parts of S. cervisiaes, promptly get the liquid dosage form probiotics, living bacteria count reaches 74 * 10
8CFU/mL.
Embodiment 2: efficient compound probiotic agent-solid type
Make liquid-type compound probiotic microbial inoculum and wheat bran 1:3 mixing by embodiment 1 method, promptly get the solid type probiotics preparation after 45 ℃ of dryings, living bacteria count reaches 67 * 10
8CFU/mL.
Embodiment 3: the fryer feeding experiment
23158 of 1 age in days AA meat chicks are selected in experiment for use, are divided into two groups of test group, control groups at random.Chicken group basal diet is identical, presses the normal procedure immunity inoculation.11610 of control groups add antibiotic routinely in the basal diet of feeding, 11548 of test group are added the efficient probiotics preparation that 1wt% embodiment 1 makes in the basal diet.Whole test is in 21 ages in days, 42 ages in days difference weighing statistics.Result of the test is: test group chick livability 92.21%, control group are 90.99%, and survival rate improves 1.22%; Average material anharmonic ratio, test group 2.027, control group 2.161, the material anharmonic ratio descends 0.134, and the test group feed conversion rate improves.
Embodiment 4: the piggy feeding experiment
120 of Du * length that child care delivers for sale * Dasanyuan piggy are selected in experiment for use, according to sex ratio, principle that body weight is close, are divided into A, B, four groups of C, D.Wherein the A group is control group, and 30 of test pig are not established repetition, and B, C, D group are respectively established two repetitions for test group, 15 every group, and male and female half and half, numbering separately.Each organizes basal diet trophic level unanimity, 30 days experimental periods.A organizes and feeds basal diet and add antibiotic routinely, the B group is added the S. cervisiae that 0.2wt% embodiment 2 fermented and cultured obtain in basal diet, the C group is added the Streptomyces thermophilus agent that 0.4wt% embodiment 2 fermented and cultured obtain in basal diet, the D group is added the efficient probiotics preparation of 1% embodiment, 2 preparations in basal diet.Result of the test shows that average daily gain is respectively, A group 581 grams, and B group 604 grams, C group 617 grams, D group 658 grams, test group D, C, B group Duo 77 grams, 36 grams, 23 grams than the control group A group respectively, and the D group is organized significant difference (P＜0.05) to A.
Embodiment 5: the milk cow feeding experiment
83 of the holstein cows that age, parity are selected in test for use, the age in days of giving milk, the output of milk are close are divided into two groups at random, 42 of experimental group, and 41 of control groups, the basal diet unanimity, 7 days preliminary trial periods,, 42 days experimental periods,, the mode of feeding is consistent with management method.Control group fed basal diet, test group add the efficient probiotics preparation of 1wt% embodiment 1 preparation in feeding basal diet, lactation amount every day of every ox of weighing record.Result of the test shows, 30.20 kilograms of average every ox outputs of milk every day of full phase of control group, and test group is 31.27.
Claims (7)
1. feeding efficient compound probiotic agent and preparation method is characterized in that the compound probiotic agent is to be mixed in proportion after the fermented and cultured respectively by bacillus licheniformis (Bacillus licheniformis), Streptomyces thermophilus (Streptococcus thermophilus), S. cervisiae (Saccharomyces cerevisiae Hansen).
2. compound probiotic agent as claimed in claim 1 is characterized in that, described bacillus licheniformis formulation preparation method is as follows:
On the sterilized liquid culture medium, insert the bacillus licheniformis of activation by 2% inoculum concentration, 37 ℃ of aerobic fementations 24 hours can get the bacillus licheniformis preparation;
The mass volume ratio of described fluid nutrient medium nutritional labeling is: 2% starch, 1% soya-bean cake hydrolyzate, 2% corn steep liquor, 1% wheat bran, dipotassium hydrogen phosphate 0.25%, 0.1%, 121 ℃ of sterilization of potassium dihydrogen phosphate 30 minutes;
Described bacillus licheniformis, aerobic growth is cultivated 30 ~ 31 ℃ of thermophilics; Can secrete bacitracin, have broad spectrum antibiotic activity, can kill pathogens such as Escherichia coli, salmonella in the animal intestinal; Can extracellular proteinase, lipase, amylase etc., promote the decomposition of macromolecular substances such as protein in the feed to absorb; Can be heat-resisting, acidproof, under the feed process neutralizes the environment of hydrochloric acid in gastric juice, bile salt in animal body, can keep higher physiological activities, cultivate easily.
3. compound probiotic agent as claimed in claim 1 is characterized in that, described Streptomyces thermophilus formulation preparation method is as follows:
In the fluid nutrient medium of sterilization, insert Streptomyces thermophilus with mass percent 5% inoculum concentration, 37 ℃ of static cultivations 24 hours can get the Streptomyces thermophilus preparation;
Described fluid nutrient medium nutriment volume ratio is: 40% Fructus Hordei Germinatus hydrolyzate, and 10% soybean hydrolyzate, 10% inorganic salt solution II was sterilized 30 minutes for 121 ℃;
Described inorganic salt solution II is meant and contains sodium acetate 5g, ammonium citrate 2g, dipotassium hydrogen phosphate 2g, magnesium sulfate 0.58g, manganese sulfate 0.25g in the 1000ml solution;
Described Streptomyces thermophilus, anaerobism or facultative aerobic, 40 ~ 42 ℃ of growth temperatures, pH value 5.5 ~ 6.2; Utilize the breeding of nourishing and growing in the enteron aisle in animal body,, suppress the pathogen growth and breeding with pathogen competition nutriment; Can secrete nisalpin, suppress and kill pathogen; Metabolism produces lactic acid, and acetate promotes the absorption of vitamin D, calcium, iron ion in the feed, has additional nutrients.
4. compound probiotic agent as claimed in claim 1 is characterized in that, described S. cervisiae bacteria preparation Preparation Method is as follows:
On the solid medium of sterilization, insert the S. cervisiae of activation with 10% inoculum concentration, 30 ~ 32 ℃ of solid fermentations 36 ~ 48 hours, middle stirring 2 ~ 3 times can get the saccharomycete preparation;
Described solid medium nutritional labeling percentage is:: 40% pomace, 10% wheat bran, 40% malt root, 10% inorganic salt solution I, an amount of water mixes, and sterilizes 30 minutes for 121 ℃;
Described inorganic salt solution I is: contain ammonium sulfate 40g, dipotassium hydrogen phosphate 0.2g, potassium dihydrogen sulfate 0.8g, calcium chloride 0.02g, magnesium sulfate 0.08g in the 1000ml solution;
Described S. cervisiae bacterium protein content height can tolerate hydrochloric acid in gastric juice and bile salt in the animal intestinal and survives; In animal intestinal, can secrete the sanguinariness factor, kill the pathogen in the enteron aisle.
5. compound probiotic agent as claimed in claim 1, it is characterized in that, the probio ratio is in the described compound probiotic agent: 30 ~ 50 parts in bacillus licheniformis preparation, and 30 ~ 50 parts in Streptomyces thermophilus preparation, the saccharomyces cerevisiae bacteria preparation mixes the liquid-type microbial inoculum for 10 ~ 40 parts; With wheat bran or rice husk absorption, air-dry or 40 ~ 50 ℃ of dryings can get probio solid type microbial inoculum.
6. as claims 1 described compound probiotic agent, it is characterized in that effective microbe content is 8 ~ 120 * 10 in the described compound probiotic agent
8CFU/mL or 6 ~ 150 * 10
8CFU/g.
7. as claims 1 described compound probiotic agent, it is characterized in that the dosage that described compound probiotic agent can 0.1 ~ 0.5% adds in animal and fowl fodder or the drinking water and uses.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103468630A (en) * | 2013-10-09 | 2013-12-25 | 长沙理工大学 | Method for producing probiotics spore powder by means of solid state fermentation |
| CN103952336A (en) * | 2014-03-28 | 2014-07-30 | 内蒙古和美科盛生物技术有限公司 | Bacillus licheniformis-Bacillus subtilis-Lactobacillus casei preparation and preparation method thereof |
| CN108967693A (en) * | 2018-07-09 | 2018-12-11 | 中国农业科学院北京畜牧兽医研究所 | A kind of weanling pig mixed feed and its application containing compound probiotic |
| CN113769422A (en) * | 2021-07-19 | 2021-12-10 | 孙元力 | Plant fiber adsorption drying method for probiotics |
-
2013
- 2013-04-23 CN CN 201310141668 patent/CN103211085A/en active Pending
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103468630A (en) * | 2013-10-09 | 2013-12-25 | 长沙理工大学 | Method for producing probiotics spore powder by means of solid state fermentation |
| CN103468630B (en) * | 2013-10-09 | 2015-09-23 | 长沙理工大学 | A kind of solid state fermentation produces the method for probiotic bacterium spore powder |
| CN103952336A (en) * | 2014-03-28 | 2014-07-30 | 内蒙古和美科盛生物技术有限公司 | Bacillus licheniformis-Bacillus subtilis-Lactobacillus casei preparation and preparation method thereof |
| CN103952336B (en) * | 2014-03-28 | 2016-06-29 | 内蒙古和美科盛生物技术有限公司 | Bacillus licheniformis, bacillus subtilis and lactobacillus casei preparation and preparation |
| CN108967693A (en) * | 2018-07-09 | 2018-12-11 | 中国农业科学院北京畜牧兽医研究所 | A kind of weanling pig mixed feed and its application containing compound probiotic |
| CN113769422A (en) * | 2021-07-19 | 2021-12-10 | 孙元力 | Plant fiber adsorption drying method for probiotics |
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Application publication date: 20130724 |