CN101836688B - Preparation method of antibiotic-free microbial fermentation feed - Google Patents
Preparation method of antibiotic-free microbial fermentation feed Download PDFInfo
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- CN101836688B CN101836688B CN200910064844A CN200910064844A CN101836688B CN 101836688 B CN101836688 B CN 101836688B CN 200910064844 A CN200910064844 A CN 200910064844A CN 200910064844 A CN200910064844 A CN 200910064844A CN 101836688 B CN101836688 B CN 101836688B
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Abstract
The invention discloses a preparation method of antibiotic-free microbial fermentation feed, which is used for producing green fermentation feed added with no chemical. The strain formula of a lactobacillus fermenting agent thereof comprises 30 percent of issatchenkia orientalis, 30 percent of abnormal pichia pastoris and 40 percent of pediococcus pentosaceus; the preparation method thereof comprises the following steps: carrying out amplification and culture for test tube strains respectively through a triangular flask and a fermentation tank, then mixing according to a certain proportion to prepare a liquid lactobacillus fermenting agent and a solid lactobacillus fermenting agent, and then adding the lactobacillus fermenting agents into the feed and fermenting into the antibiotic-free microbial fermentation feed. Because acidic substances are generated by adding beneficial strains capable of excreting a large amount of various digestive enzymes, the invention decreases the PH value, provides an anaerobic environment for lactobacilli, is beneficial to the field planting of the lactobacilli in intestines, can effectively improve the microeubiosis of animal intestines and thins nutritional factors, thereby improves the immunity of animals and has the actions of increasing the meat-feed ratio, having no medicament residue and improving the meat quality.
Description
Technical field: the present invention relates to a kind of animal and fowl fodder, specifically is to produce a kind of preparation method who does not contain antibiotic fermentative feedstuff of microbe who does not add the green fermented feed of chemicals.
Background technology: the Along with people's growth in the living standard, people more and more pay attention to food-safety problem.Especially a series of food-safety problems of melamine incident initiation make people recognize the importance of food security more.As the source of animal food, feed safety is exactly a food security.In pig feed and chicken feed, generally be added with antibiotic and hormone at present, cause the pork produced and chicken and egg mouthfeel poor, antibiotic content exceeds standard, and it is healthy to influence people.In order to guarantee the quality of meat, many producers also attempt producing and do not add antibiotic fermented feed, but because the bacterial classification of fermentation usefulness and the restriction of zymotechnique make this antibiotic-free feed be difficult to develop in a healthy way.
Summary of the invention: the invention provides a kind of immunity and growth performance that can improve live pig, milk cow, chicken, a kind of preparation method who does not contain antibiotic fermentative feedstuff of microbe of simple and easy low-cost production.Its lactic acid bacteria fermenting agent bacterial classification prescription is: Issatchenkia orientalis bacterium (Latin formal name used at school: Issatchenkia orientalis; Depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center; Preservation date: on December 18th, 2008; CGMCC No:2813) 30%, unusual Pichia yeast (Latin formal name used at school: Pichia anomala depositary institution is called for short and numbering:; Depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center; Preservation date: on December 18th, 2008; Depositary institution is called for short and numbering: CGMCC No:2814) 30% and Pediococcus pentosaceus (Latin formal name used at school: Pediococcus pento saceus; Depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center; Preservation date: on December 18th, 2008; Depositary institution is called for short and numbering: CGMCC No:2815) 40%; Its preparation method is to use triangular flask and fermentation tank amplification cultivation to above-mentioned test tube strains respectively; Be mixed with into by a certain percentage liquid lactic acid bacteria fermenting agent and solid lactic acid bacterium leavening agent again, add lactic acid bacteria fermenting agent to be fermented in the feed to then and do not contain antibiotic fermentative feedstuff of microbe.Because the present invention utilizes the beneficial microbe fermented feed, through adding the Issatchenkia orientalis bacterium that can secrete various digestive ferments in a large number, the unusual Pichia yeast that protein content is high produces acidic materials and reduces pH value; For Pediococcus pentosaceus provides anaerobic environment, help lactic acid bacteria field planting in enteron aisle, can effectively improve the animal intestinal microecological balance; The refinement trophic factors, thus the immunity of animal improved, and have the raising meat feed ratio; Drug residue free improves the effect of meat.
The present invention is used for three kinds of probios of fermented feed and produces with the method for fermentation tank amplification cultivation respectively, produces the lactic acid bacteria fermenting agent inoculating proportion of fermentative feedstuff of microbe and does suitable adjustment according to different animals or with the different growing stages of a kind of animal.It is carrier that the used solid lactic acid bacterium leavening agent of the present invention adopts corn, wheat bran etc., adheres in order to microorganism fungus kind, and its useful viable bacteria content can reach 1,000,000,000/gram.Beneficial bacterium content can reach 2,000,000,000/milliliter in the used liquid lactic acid bacteria fermenting agent of the present invention.Result of the test through in recent years shows that fermentative feedstuff of microbe improves non-specific immunity, the positive effect of having increased income to improving the live pig meat feed ratio.Live pig searches for food behind the feed that ferments, and fur is smooth, the colour of skin is ruddy, the strong stern of the back of the body is fertile, the state of mind is good, and lactic acid bacteria fermenting agent can substitute the antibiotic in the feed fully.Compare with using antibiotic traditional feeding manner, use the live pig average daily gain of the fermentative feedstuff of microbe group of antibiotic-free to improve 5 to 6 percentage points.From Economic and Efficiency Analysis, the fermentative feedstuff of microbe of antibiotic-free is increased income about 15 yuan than average every the pig of traditional antibiotic mixed fodder group.
Culture presevation brief description in the lactic acid bacteria fermenting agent prescription of the present invention:
1, Issatchenkia orientalis bacterium: Latin formal name used at school: Issatchenkia orientalis;
Depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center;
Depositary institution address: Datun Road, Chaoyang District, Beijing City;
Preservation date: on December 18th, 2008;
Depositary institution is called for short and numbering: CGMCC No:2813.
2, unusual Pichia yeast: Latin formal name used at school: Pichia anomala;
Depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center;
Depositary institution address: Datun Road, Chaoyang District, Beijing City;
Preservation date: on December 18th, 2008;
Depositary institution is called for short and numbering: CGMCC No:2814.
3, Pediococcus pentosaceus: Latin formal name used at school: Pediococcus pento saceus;
Depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center;
Depositary institution address: Datun Road, Chaoyang District, Beijing City;
Preservation date: on December 18th, 2008;
Depositary institution is called for short and numbering: CGMCC No:2815.
The specific embodiment: embodiment 1: the bacterial classification prescription of lactic acid bacteria fermenting agent is: Issatchenkia orientalis bacterium (CGMCC No:2813) 30%, unusual Pichia yeast (CGMCC No:2814) 30% and Pediococcus pentosaceus (CGMCC No:2815) 40%; Its preparation method is with triangular flask and fermentation tank culture amplification the test tube seed; Be mixed with by a certain percentage again and become not contain antibiotic liquid lactic acid bacteria fermenting agent, solid lactic acid bacterium leavening agent, add lactic acid bacteria fermenting agent to be fermented in the feed to then and do not contain antibiotic fermentative feedstuff of microbe.
Embodiment 2: the triangular flask culture medium prescription and the cultural method of lactic acid bacteria fermenting agent are:
Issatchenkia orientalis bacterium CGMCC No:2813: malt extract medium (12 ° of B é), pH5.5～6.5,121 ℃ sterilization 20min; Its inoculum concentration is 1%, cultivates 30h down in 28 ℃;
Unusual Pichia yeast CGMCC No:2814:. malt extract medium (12 ° of B é), pH5.5～6.5,121 ℃ sterilization 20min; Its inoculum concentration is 1%, cultivates 30h down in 28 ℃;
Pediococcus pentosaceus CGMCC No:2815: caseinhydrolysate 10.0g, yeast extract 5.0g, glucose 20.0g, Tween 80 1.0g, ammonium citrate 2.0g, dipotassium hydrogen phosphate 2.0g, sodium acetate 2.0g, MnSO
4.7H
2O 0.2g, MgSO
40.05g, distilled water 1000ml, pH6.2～6.5,121 ℃ sterilization 20min; Its inoculum concentration is 1%, cultivates 24h down in 35 ℃.
Embodiment 3: the fermentation tank culture based formulas and the cultural method of lactic acid bacteria fermenting agent are:
Issatchenkia orientalis bacterium CGMCC No:2813: malt extract medium (12 ° of B é), 5.5～6.5,121 ℃ of sterilizations of pH 25min; Or: brown sugar 20g, soybean sprout juice 50ml, dipotassium hydrogen phosphate 2g, water 1000ml, pH5.5～6.5,121 ℃ sterilization 25min; Its inoculum concentration is 3%, in 28 ℃, and pH5.5～6.5 time cultivation 30h;
Unusual Pichia yeast CGMCC No:2814: malt extract medium (12 ° of B é), pH 5.5～6.5,121 ℃ of sterilization 25min; Or: brown sugar 20g, soybean sprout juice 50ml, dipotassium hydrogen phosphate 2g, water 1000ml, pH5.5～6.5,121 ℃ sterilization 25min; Its inoculum concentration is 3%, in 28 ℃, and pH5.5～6.5 time cultivation 30h; Pediococcus pentosaceus CGMCC No:2815: brown sugar 20g, the 50ml of Tomato juice, yeast extract 5g, dipotassium hydrogen phosphate 2g, Tween 80 0.5ml, pH 6.2～6.5,121 ℃ of sterilization 20min; Its inoculum concentration is 3%, in 35 ℃, and pH5.5～6.5 time cultivation 24h.
Embodiment 4: the solid culture based formulas and the cultural method of lactic acid bacteria fermenting agent are:
Issatchenkia orientalis bacterium CGMCC No:2813: corn flour 45%, dregs of beans 25%, wheat bran 25%, brown sugar 5%, pH 5.5～6.5,121 ℃ of sterilization 60min; Its inoculum concentration is 6%, and in 28 ℃, pH cultivates 32h 5.5～6.5 times;
Unusual Pichia yeast CGMCC No:2814: corn flour 45%, dregs of beans 25%, wheat bran 25%, brown sugar 5%, pH 5.5～6.5,121 ℃ of sterilization 60min; Its inoculum concentration is 6%, and in 28 ℃, pH cultivates 32h 5.5～6.5 times;
Pediococcus pentosaceus CGMCC No:2815: corn flour 45%, dregs of beans 25%, wheat bran 25%, brown sugar 5%, pH 6.5～7.3,121 ℃ of sterilization 60min; Its inoculum concentration is 6%, and in 35 ℃, pH cultivates 32h 5.5～6.5 times.
Embodiment 5: the preparation technology of liquid lactic acid bacteria fermenting agent is: test tube strains → triangular flask one-level kind → fermentation tank secondary kind → hybrid combination → sterile filling → finished product; Its hybrid combination bacterial classification ratio is: Issatchenkia orientalis bacterium (CGMCC No:2813) 30%, unusual Pichia yeast (CGMCC No:2814) 30%, Pediococcus pentosaceus (CGMCC No:2815) 40%.
The liquid lactic acid bacteria fermenting agent adds the method for producing fermentative feedstuff of microbe in the feed to:
(1) dilution of lactic acid bacteria fermenting agent: the ratio according to 1: 10 is diluted, and soon 1L liquid lactic acid bacteria fermenting agent is poured in the 10kg water and stirred;
(2) fermentation: the liquid lactic acid bacteria fermenting agent that the 10kg dilution is good adds in the 100kg pig feed, and it is even with mixer or manual stirring to add water.The feed that stirs is contained in the ventilative woven bag, keeps feed fluffy ventilative, place the 24-48h that ferments in 28-35 ℃ the room.
Embodiment 6: the preparation technology of solid lactic acid bacterium leavening agent is: test tube strains → triangular flask one-level kind → three grades of kinds of fermentation tank secondary kind → solid culture → aseptic airing → hybrid combination → finished product; Its hybrid combination bacterial classification ratio is: Issatchenkia orientalis bacterium (CGMCC No:2813) 30%, unusual Pichia yeast (CGMCC No:2814) 30%, Pediococcus pentosaceus (CGMCC No:2815) 40%; Issatchenkia orientalis bacterium CGMCC No:2813 is an Aluminum Drum with unusual Pichia yeast CGMCC No:2814 solid fermentation container, and Pediococcus pentosaceus CGMCC No:2815 solid fermentation container is the sealed plastic bucket; Solid spawn after the fermentation is dried in order to avoid pollute in 6-12h fast.
The solid lactic acid bacterium leavening agent adds the method for producing fermentative feedstuff of microbe in the feed to:
(1) activation of solid lactic acid bacterium leavening agent: take the amount of 1kg solid lactic acid bacterium leavening agent by the 1t feed, under 40 ℃ of left and right sides conditions, the solid lactic acid bacterium leavening agent is placed on soaks activation 2-3h in the 10kg warm water;
(2) fermentation: the lactic acid bacteria fermenting agent that activation is good adds in the 100kg feed; It is even with mixer or manual stirring to add water; Again the feed that stirs is contained in the ventilative woven bag, keeps feed fluffy ventilative, place the 24-48h that ferments in 28-35 ℃ the room.
Embodiment 7: the present invention utilizes the fermentative feedstuff of microbe of lactic acid bacteria fermenting agent production to have special wine flavour, and useful viable count is more than 1,000,000,000/gram.The fermentative feedstuff of microbe that ferments should use up within 6 months.Should note in preparation with in using:
(1) moisture content requires: stirring back moisture content is 55%-65%, does not promptly seep water with having held agglomerating, and the agglomerate of loosing one's grip can split, and hand is rubbed then and loose.Water quality requirement: the required water of feed fermentation does not need extra sterilization, but should be clean running water or underground water.
(2) temperature requirement: the required temperature range of fermenting is 20-40 ℃, is lower than 20 ℃ of difficult fermentations.
(3) ventilation requirement: the woven bag of splendid attire feed should select not have the sack of the good permeability of inner membrance, and charging can not be real excessively, and the water content of feed will be controlled between the 55%-65% simultaneously, to guarantee to have an amount of oxygen supply.
(4) fermentation time: different according to environment temperature, water content and throughput, required time of feed fermentation is also different, and according to standard operation, fermenting can reach requirement in 24-48 hour.
(5) requirement of fermentation raw material: the feed that must select not contain additives such as antibiotic, hormone, anticorrisive agent, pigment, clenbuterol hydrochloride is as fermentation raw material.
Claims (1)
1. preparation method who does not contain antibiotic fermentative feedstuff of microbe is characterized in that its preparation method is:
(1) the lactic acid bacteria fermenting agent bacterial classification prescription of production fermentative feedstuff of microbe is: Issatchenkia orientalis bacterium (Issatchenkia orientalis; CGMCC No:2813) 30% unusual Pichia yeast (Pichia anomala; CGMCC No:2814) 30% and Pediococcus pentosaceus (Pediococcus pentosaceus; CGMCC No:2815) 40%; Its preparation method is to increase above-mentioned test tube strains with triangular flask and fermentation tank culture respectively earlier; Be mixed with into by a certain percentage liquid lactic acid bacteria fermenting agent and solid lactic acid bacterium leavening agent again, then liquid lactic acid bacteria or solid lactic acid bacterium leavening agent added to be fermented in the feed and do not contain antibiotic fermentative feedstuff of microbe;
(2) preparation technology of its liquid lactic acid bacteria fermenting agent is: test tube strains → triangular flask one-level kind → fermentation tank secondary kind → hybrid combination → sterile filling → finished product; Its hybrid combination bacterial classification ratio is: Issatchenkia orientalis bacterium (CGMCC No:2813) 30%, unusual Pichia yeast (CGMCC No:2814) 30%, Pediococcus pentosaceus (CGMCC No:2815) 40%;
(3) preparation technology of its solid lactic acid bacterium leavening agent is: test tube strains → triangular flask one-level kind → three grades of kinds of fermentation tank secondary kind → solid culture → aseptic airing → hybrid combination → finished product; Its hybrid combination bacterial classification ratio is: Issatchenkia orientalis bacterium (CGMCC No:2813) 30%, unusual Pichia yeast (CGMCC No:2814) 30%, Pediococcus pentosaceus (CGMCC No:2815) 40%; Issatchenkia orientalis bacterium CGMCC No:2813 is an Aluminum Drum with unusual Pichia yeast CGMCC No:2814 solid fermentation container, and Pediococcus pentosaceus CGMCC No:2815 solid fermentation container is the sealed plastic bucket; Solid spawn after the fermentation is dried in 6-12h in order to avoid pollute fast;
(4) its liquid lactic acid bacteria fermenting agent adds in the feed method of producing fermentative feedstuff of microbe to and is:
1. the dilution of lactic acid bacteria fermenting agent: the ratio according to 1: 10 is diluted, and soon 1L liquid lactic acid bacteria fermenting agent is poured in the 10kg water and stirred;
2. fermentation: the liquid lactic acid bacteria fermenting agent that the 10kg dilution is good adds in the 100kg pig feed, and it is even with mixer or manual stirring to add water; The feed that stirs is contained in the ventilative woven bag, keeps feed fluffy ventilative, place the 24-48h that ferments in 28-35 ℃ the room;
(5) its solid lactic acid bacterium leavening agent adds in the feed method of producing fermentative feedstuff of microbe to and is:
1. the activation of solid lactic acid bacterium leavening agent: take the amount of 1kg solid lactic acid bacterium leavening agent by the 1t feed, under 40 ℃ of left and right sides conditions, the solid lactic acid bacterium leavening agent is placed on soaks activation 2-3h in the 10kg warm water;
2. fermentation: the lactic acid bacteria fermenting agent that activation is good adds in the 100kg feed; It is even with mixer or manual stirring to add water; Again the feed that stirs is contained in the ventilative woven bag, keeps feed fluffy ventilative, place the 24-48h that ferments in 28-35 ℃ the room;
(6) the triangular flask culture medium prescription of its lactic acid bacteria fermenting agent and cultural method are:
The malt extract medium of Issatchenkia orientalis bacterium CGMCC No:2813:12 Baume degrees, pH5.5～6.5,121 ℃ sterilization 20min; Its inoculum concentration is 1%, cultivates 30h down in 28 ℃;
The malt extract medium of unusual Pichia yeast CGMCC No:2814:12 Baume degrees, pH5.5～6.5,121 ℃ sterilization 20min; Its inoculum concentration is 1%, cultivates 30h down in 28 ℃;
Pediococcus pentosaceus CGMCC No:2815: caseinhydrolysate 10.0g, yeast extract 5.0g, glucose 20.0g, Tween 80 1.0g, ammonium citrate 2.0g, dipotassium hydrogen phosphate 2.0g, sodium acetate 2.0g, MnSO
47H
2O 0.2g, MgSO
40.05g, distilled water 1000ml, pH6.2～6.5,121 ℃ sterilization 20min; Its inoculum concentration is 1%, cultivates 24h down in 35 ℃;
(7) the solid culture based formulas of its lactic acid bacteria fermenting agent and cultural method are:
Issatchenkia orientalis bacterium CGMCC No:2813: corn flour 45%, dregs of beans 25%, wheat bran 25%, brown sugar 5%, pH 5.5～6.5,121 ℃ of sterilization 60min; Its inoculum concentration is 6%, and in 28 ℃, pH cultivates 32h 5.5～6.5 times;
Unusual Pichia yeast CGMCC No:2814: corn flour 45%, dregs of beans 25%, wheat bran 25%, brown sugar 5%, pH 5.5～6.5,121 ℃ of sterilization 60min; Its inoculum concentration is 6%, and in 28 ℃, pH cultivates 32h 5.5～6.5 times;
Pediococcus pentosaceus CGMCC No:2815: corn flour 45%, dregs of beans 25%, wheat bran 25%, brown sugar 5%, pH 6.5～7.3,121 ℃ of sterilization 60min; Its inoculum concentration is 6%, and in 35 ℃, pH cultivates 32h 5.5～6.5 times.
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Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102106487B (en) * | 2011-02-01 | 2012-12-05 | 大连韩伟养鸡有限公司 | Biological fermentation feed for laying chickens and production method |
| CN102488120B (en) * | 2011-12-28 | 2013-02-13 | 广西东园生态农业科技有限公司 | Preparation method of fermenting ensilaged feed |
| CN102972655B (en) * | 2012-11-30 | 2015-03-25 | 来光业 | Preparation method of microbial fermented feed without antibiotic for sow |
| CN102972652B (en) * | 2012-11-30 | 2015-03-25 | 来光业 | Preparation method of microbial fermented feed without antibiotic for pork pig |
| CN105961845A (en) * | 2016-05-06 | 2016-09-28 | 江苏盐城源耀生物科技有限公司 | New antibiotic substitute and application thereof in livestock and poultry breeding |
| CN107937286B (en) * | 2017-12-08 | 2020-11-06 | 西南大学 | Pichia anomala, fermentation culture of pichia anomala and application of pichia anomala |
| CN113662090A (en) * | 2021-08-27 | 2021-11-19 | 江苏优仕生物科技发展有限公司 | Production process and production equipment of microbial fermented feed for replacing antibiotics |
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