CN102660534A - Bacillus subtilis preparation as well as preparation method and application thereof - Google Patents
Bacillus subtilis preparation as well as preparation method and application thereof Download PDFInfo
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- CN102660534A CN102660534A CN2012101395470A CN201210139547A CN102660534A CN 102660534 A CN102660534 A CN 102660534A CN 2012101395470 A CN2012101395470 A CN 2012101395470A CN 201210139547 A CN201210139547 A CN 201210139547A CN 102660534 A CN102660534 A CN 102660534A
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Abstract
The invention discloses a bacillus subtilis preparation as well as a preparation method and application thereof, belonging to the field of probiotics. The bacillus subtilis preparation is prepared from bacillus subtilis BS0903 and a vector; the content of viable organisms of the bacillus subtilis in the preparation is 1*10<9>cfu/g<-1>*10<11>cfu/g; the bacillus subtilis BS0903 was transported to CCTCC (China General Microbiological Culture Collection Center) for preserving on March14th, 2012; and the strain number is BS0903 and the preservation number is CCTCC NO:M2012078. The bacillus subtilis preparation is prepared by adopting the following steps of: thawing and passing a bacillus subtilis strain to obtain seed broth; carrying out enriched culture on the seed broth through a fermentation tank to obtain fermentation tank broth; and mixing the fermentation tank broth with the vector and then carrying out centrifuging, spraying and drying to obtain the bacillus subtilis preparation. The bacillus subtilis preparation disclosed by the invention has the advantages of simpleness, reasonability, stable production, low cost and convenience for storage and transportation. After the bacillus subtilis preparation is used for feeding poultry through oral path (drinking water or mixing materials), the bacillus subtilis preparation has the functions of improving the digestion function, increasing the feed conversion rate and promoting the growth and is a microbiological preparation with more ideal effect and capability of improving the growth performance of the poultry.
Description
Technical field
The present invention relates to a kind of microbial preparation and preparation method thereof and application, particularly bacillus subtilis formulation and preparation method thereof and application, belong to the microorganism fodder field.
Background technology
The bacillus category microbe additive is a kind of active bacteria formulation that utilizes the useful genus bacillus among the normal microflora member to process, and has advantages such as natural high temperature resistant and acid and alkali-resistance.Subtilis is one type of aerobic growth, can forms the gram-positive microorganism of gemma, also is to generally acknowledge both at home and abroad in feed, directly to add the probiotic bacterium of using.Subtilis exists with the statospore form in preparation; Spore can be sprouted the very strong proteolytic enzyme of justacrine activity, lypase, glycase etc. rapidly after getting into animal intestinal; Some complicated compound in the feed helps to degrade; Produce useful meta-bolites, promote digesting and assimilating of nutritive substance, reach promotion growth of animal and the purpose of improving food conversion ratio.Because subtilis has the active and advantages such as promotes growth, suitable processing storage tolerance of abundant multifunctional enzyme, thereby more and more receives correlative study personnel's attention.
Summary of the invention
The object of the present invention is to provide a kind of by single bacterial strain production, low production cost, the application that is used to promote the poultry feed conversion rate, improves production performance obvious results bacillus subtilis formulation and preparation method thereof and said preparation.
The objective of the invention is to realize through following technical scheme: bacillus subtilis formulation, it is characterized in that said preparation is made by subtilis BS0903 and carrier, the subtilis viable bacteria content in the preparation is 1 * 10
9Cfu/g～1 * 10
11Cfu/g, said subtilis BS0903 (Bacillus subtilis BS0903) sent Chinese typical culture collection center preservation on March 14th, 2012, strain number: BS0903, deposit number are CCTCC NO:M2012078.
Process kind of daughter bacteria liquid after described Bacillus subtilis strain recovery is gone down to posterity and after fermentor tank increases the bacterium cultivation, obtain fermentor tank bacterium liquid, the weight percent of fermentor tank bacterium liquid and carrier is: fermentor tank bacterium liquid 80～92%, carrier 8～20%.
Described carrier is a kind of or any several kinds mixtinite in starch, talcum powder, lime carbonate, the kaolin.
The BS0903 seed culture fluid is made up of Zulkovsky starch, beef powder, NaCl, water, and its pH value is 7.2～7.4, and the weight percent of each component is: Zulkovsky starch 3%, beef powder 0.5%, NaCl0.5%, surplus are water.
The fermentor cultivation liquid that said fermentor tank increases the bacterium cultivation is made up of Zulkovsky starch, bean powder, yeast powder, peptone, potassium hydrogenphosphate, potassium primary phosphate, water; Its pH value is 7.8～8.0, and the weight percent of each component is: Zulkovsky starch 5%, bean powder 2%, yeast powder 0.8%, peptone 0.5%, potassium hydrogenphosphate 0.2%, potassium primary phosphate 0.1%, surplus are water.
The preparation method of bacillus subtilis formulation is characterized in that, may further comprise the steps:
1) placing seed culture fluid to recover subtilis BS0903 goes down to posterity; Make kind of a daughter bacteria liquid; Said subtilis BS0903 (Bacillus subtilis BS0903); Sent Chinese typical culture collection center preservation on March 14th, 2012, strain number: BS0903, deposit number are CCTCC NO:M2012078;
2) the kind daughter bacteria liquid that step 1) is made places fermentor cultivation liquid to carry out fermentor tank to increase bacterium and cultivate, make fermentor tank bacterium liquid; Said fermentor cultivation liquid is made up of Zulkovsky starch, bean powder, yeast powder, peptone, potassium hydrogenphosphate, potassium primary phosphate, water; Its pH value is 7.8～8.0, and the weight percent of each component is: Zulkovsky starch 5%, bean powder 2%, yeast powder 0.8%, peptone 0.5%, potassium hydrogenphosphate 0.2%, potassium primary phosphate 0.1%, surplus are water.
3) fermentor tank bacterium liquid and carrier are mixed, through centrifugal spray drying, make bacillus subtilis formulation, the viable bacteria content in the said bacillus subtilis formulation is 1 * 10
9Cfu/g～1 * 10
11Cfu/g, the weight percent of described fermentor tank bacterium liquid and carrier is: fermentor tank bacterium liquid 80～92%, carrier 8～20%.
The described carrier of step 3) is a kind of or any several kinds molectron in starch, talcum powder, lime carbonate, the kaolin.
Seed culture fluid described in the step 1) is made up of Zulkovsky starch, beef powder, NaCl, water, and its pH value is 7.2～7.4, and the weight percent of each component is: Zulkovsky starch 3%, beef powder 0.5%, NaCl0.5%, surplus are water.
The application of above-mentioned bacillus subtilis formulation is characterized in that, the addition of bacillus subtilis formulation by poultry feed or drinking-water weight 0.1～0.2% added in poultry feed or the drinking-water, can improve the transformation efficiency 13.8～21.5% of feed.
The bacillus subtilis formulation that finally makes is a dry-powdered.
The present invention has adopted a kind of subtilis BS0903 (Bacillus subtilis BS0903) bacterial strain; This bacterial strain is in the healthy poultry enteron aisles of some strains of collecting over the years, preserving the isolating bacillus subtilis strain, and a strain that after repeated screening, obtains has the Bacillus subtilis strain of strong cellulase, glycase and alkaline protease activity.
Advantages of simple of the present invention, science are produced and are stablized, and cost is low, preserve and convenient transportation.Behind bacillus subtilis formulation oral route (drinking-water or the spice) poultry of feeding, have the digestive function of improvement, improve food conversion ratio, promote the effect of growth.Use through regional testing and to show, this bacillus subtilis formulation is converted water or spice after, the poultry of feeding, average weight gain reaches 13.8～21.5% as a result, the present invention is the microbial preparation that a kind of comparatively ideal improves the poultry growth performance.
Embodiment
Embodiment one
Bacillus subtilis formulation, said preparation is made by subtilis BS0903 and carrier, and the weight percent of fermentor tank bacterium liquid and carrier is: fermentor tank bacterium liquid 80～92%, carrier 8～20%.Carrier is a kind of or any several kinds mixtinite in starch, talcum powder, lime carbonate, the kaolin.Subtilis viable bacteria content in the preparation is 1 * 10
9Cfu/g～1 * 10
11Cfu/g, subtilis BS0903 (Bacillus subtilis BS0903) sent Chinese typical culture collection center preservation on March 14th, 2012, strain number: BS0903, deposit number are CCTCC NO:M2012078.
Process kind of daughter bacteria liquid after the Bacillus subtilis strain recovery is gone down to posterity and after fermentor tank increases the bacterium cultivation, obtain fermentor tank bacterium liquid; The BS0903 seed culture fluid is made up of Zulkovsky starch, beef powder, NaCl, water; Its pH value is 7.2～7.4, and the weight percent of each component is: Zulkovsky starch 3%, beef powder 0.5%, NaCl0.5%, surplus are water.The fermentor cultivation liquid that fermentor tank increases the bacterium cultivation is made up of Zulkovsky starch, bean powder, yeast powder, peptone, potassium hydrogenphosphate, potassium primary phosphate, water; Its pH value is 7.8～8.0, and the weight percent of each component is: Zulkovsky starch 5%, bean powder 2%, yeast powder 0.8%, peptone 0.5%, potassium hydrogenphosphate 0.2%, potassium primary phosphate 0.1%, surplus are water.
Embodiment two
The preparation method of bacillus subtilis formulation is characterized in that, may further comprise the steps:
1) placing seed culture fluid to recover subtilis BS0903 goes down to posterity; Make kind of a daughter bacteria liquid; Subtilis BS0903 (Bacillus subtilis BS0903); Sent Chinese typical culture collection center preservation on March 14th, 2012, strain number: BS0903, deposit number are CCTCC NO:M2012078; Seed culture fluid is made up of Zulkovsky starch, beef powder, NaCl, water, and its pH value is 7.2～7.4, and the weight percent of each component is: Zulkovsky starch 3%, beef powder 0.5%, NaCl0.5%, surplus are water;
2) the kind daughter bacteria liquid that step 1) is made places fermentor cultivation liquid to carry out fermentor tank to increase bacterium and cultivate, make fermentor tank bacterium liquid; Fermentor cultivation liquid is made up of Zulkovsky starch, bean powder, yeast powder, peptone, potassium hydrogenphosphate, potassium primary phosphate, water; Its pH value is 7.8～8.0, and the weight percent of each component is: Zulkovsky starch 5%, bean powder 2%, yeast powder 0.8%, peptone 0.5%, potassium hydrogenphosphate 0.2%, potassium primary phosphate 0.1%, surplus are water.
3) fermentor tank bacterium liquid and carrier are mixed, through centrifugal spray drying, make bacillus subtilis formulation, the viable bacteria content in the bacillus subtilis formulation is 1 * 10
9Cfu/g～1 * 10
11Cfu/g, the weight percent of fermentor tank bacterium liquid and carrier is: fermentor tank bacterium liquid 80～92%, carrier 8～20%; Carrier is a kind of or any several kinds molectron in starch, talcum powder, lime carbonate, the kaolin.
Embodiment three
Process kind of a daughter bacteria liquid after subtilis BS0903 bacterial classification gone down to posterity through recovery, will plant daughter bacteria liquid again and increase through fermentor tank that bacterium is cultivated the back and carrier mixes, through centrifugal spray drying, processing viable bacteria content is 1 * 10
9Cfu/g～1 * 10
11The cfu/g bacillus subtilis formulation.Yellow chicken 120 plumages of 7 ages in days Soviet Union fowl that body weight is close are divided into control group and test group at random, and every group 60 plumage raised separately.Control group fed basal diet, test group add the embodiment one of basal diet weight 0.15% or the bacillus subtilis formulation that embodiment two makes in basal diet.Keep term harmonizations such as test group and control group temperature, humidity, illumination, ventilation in the feeding process.Weigh at 14,21,28,35,42,49 ages in days respectively, write down body weight of every plumage chicken and the feed consumption rate of every group of chicken, the full phase of statistical test is respectively organized average weight gain and food consumption.Test group yellow-feathered broiler average weight gain 15.6% as a result.
Embodiment four
7 aa broiler chicken, 100 plumages that body weight is close are divided into control group and test group at random, and every group 50 plumage raised separately.Control group fed basal diet, test group add the embodiment one of basal diet weight 0.1% or the bacillus subtilis formulation that embodiment two makes in basal diet.Keep term harmonizations such as test group and control group temperature, humidity, illumination, ventilation in the feeding process.Weigh at 14,21,28,35,42,49 ages in days respectively, write down body weight of every plumage chicken and the feed consumption rate of every group of chicken, the full phase of statistical test is respectively organized average weight gain and food consumption.Test group AA fryer average weight gain 21.5% as a result.
Embodiment five
14 age in days Gaoyou duck 80 plumages that body weight is close are divided into control group and test group at random, and every group 40 plumage raised separately.Control group and the test group basal diet of all feeding, test group add the embodiment one of drinking-water weight 0.2% or the bacillus subtilis formulation that embodiment two makes in drinking-water.Keep term harmonizations such as test group and control group temperature, humidity, illumination, ventilation in the feeding process.Weigh at 21,28,35,42,49,56 ages in days respectively, write down body weight of every plumage duck and the feed consumption rate of every group of duck, the full phase of statistical test is respectively organized average weight gain and food consumption.Test group Gaoyou duck average weight gain 14.7% as a result.
Embodiment six
14 age in days Yangzhou goose 80 plumages that body weight is close are divided into control group and test group at random, and every group 40 plumage raised separately.Control group and the test group basal diet of all feeding, test group add the embodiment one of drinking-water weight 0.2% or the bacillus subtilis formulation that embodiment two makes in drinking-water.Keep term harmonizations such as test group and control group temperature, humidity, illumination, ventilation in the feeding process.Weigh at 28,42,56,70 ages in days respectively, write down body weight of every plumage goose and the feed consumption rate of every group of goose, the full phase of statistical test is respectively organized average weight gain and food consumption.Test group Yangzhou goose average weight gain 13.8% as a result.
Claims (9)
1. a bacillus subtilis formulation is characterized in that, said preparation is made by subtilis BS0903 and carrier, and the subtilis viable bacteria content in the preparation is 1 * 10
9Cfu/g～1 * 10
11Cfu/g, said subtilis BS0903 (Bacillus subtilis BS0903) sent Chinese typical culture collection center preservation on March 14th, 2012, strain number: BS0903, deposit number are CCTCC NO:M2012078.
2. bacillus subtilis formulation according to claim 1; It is characterized in that; Process kind of daughter bacteria liquid after described Bacillus subtilis strain recovery is gone down to posterity and after fermentor tank increases the bacterium cultivation, obtain fermentor tank bacterium liquid, the weight percent of fermentor tank bacterium liquid and carrier is: fermentor tank bacterium liquid 80～92%, carrier 8～20%.
3. bacillus subtilis formulation according to claim 1 is characterized in that, described carrier is a kind of or any several kinds mixtinite in starch, talcum powder, lime carbonate, the kaolin.
4. bacillus subtilis formulation according to claim 1; It is characterized in that; The BS0903 seed culture fluid is made up of Zulkovsky starch, beef powder, NaCl, water; Its pH value is 7.2～7.4, and the weight percent of each component is: Zulkovsky starch 3%, beef powder 0.5%, NaCl0.5%, surplus are water.
5. bacillus subtilis formulation according to claim 2; It is characterized in that; The fermentor cultivation liquid that said fermentor tank increases the bacterium cultivation is made up of Zulkovsky starch, bean powder, yeast powder, peptone, potassium hydrogenphosphate, potassium primary phosphate, water; Its pH value is 7.8～8.0, and the weight percent of each component is: Zulkovsky starch 5%, bean powder 2%, yeast powder 0.8%, peptone 0.5%, potassium hydrogenphosphate 0.2%, potassium primary phosphate 0.1%, surplus are water.
6. the preparation method of a bacillus subtilis formulation is characterized in that, may further comprise the steps:
1) placing seed culture fluid to recover subtilis BS0903 goes down to posterity; Make kind of a daughter bacteria liquid; Said subtilis BS0903 (Bacillus subtilis BS0903); Sent Chinese typical culture collection center preservation on March 14th, 2012, strain number: BS0903, deposit number are CCTCC NO:M2012078;
2) the kind daughter bacteria liquid that step 1) is made places fermentor cultivation liquid to carry out fermentor tank to increase bacterium and cultivate, make fermentor tank bacterium liquid; Said fermentor cultivation liquid is made up of Zulkovsky starch, bean powder, yeast powder, peptone, potassium hydrogenphosphate, potassium primary phosphate, water; Its pH value is 7.8～8.0, and the weight percent of each component is: Zulkovsky starch 5%, bean powder 2%, yeast powder 0.8%, peptone 0.5%, potassium hydrogenphosphate 0.2%, potassium primary phosphate 0.1%, surplus are water;
3) fermentor tank bacterium liquid and carrier are mixed, through centrifugal spray drying, make bacillus subtilis formulation, the viable bacteria content in the said bacillus subtilis formulation is 1 * 10
9Cfu/g～1 * 10
11Cfu/g, the weight percent of described fermentor tank bacterium liquid and carrier is: fermentor tank bacterium liquid 80～92%, carrier 8～20%.
7. the preparation method of bacillus subtilis formulation according to claim 6 is characterized in that, the described carrier of step 3) is a kind of or any several kinds molectron in starch, talcum powder, lime carbonate, the kaolin.
8. the preparation method of bacillus subtilis formulation according to claim 6; It is characterized in that; Seed culture fluid described in the step 1) is made up of Zulkovsky starch, beef powder, NaCl, water; Its pH value is 7.2～7.4, and the weight percent of each component is: Zulkovsky starch 3%, beef powder 0.5%, NaCl0.5%, surplus are water.
9. application like claim 1 or 6 described bacillus subtilis formulations; It is characterized in that; The addition of bacillus subtilis formulation by poultry feed or drinking-water weight 0.1～0.2% added in poultry feed or the drinking-water, can improve the transformation efficiency 13.8～21.5% of feed.
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Cited By (6)
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| CN104611257A (en) * | 2014-12-25 | 2015-05-13 | 浙江惠嘉生物科技有限公司 | Preparation method of bacillus licheniformis preparations for drinking water of livestock and poultry |
| CN104805031A (en) * | 2014-09-17 | 2015-07-29 | 德强生物股份有限公司 | Bacillus subtilis and preparation method and application thereof |
| CN107236691A (en) * | 2017-06-28 | 2017-10-10 | 四川农业大学 | A kind of beaver rabbit source bacillus subtilis and its application in growth of Rex rabbits performance is improved |
| CN108125067A (en) * | 2017-12-01 | 2018-06-08 | 杭州柏普泰生物科技有限公司 | A kind of disease preventing and treating and the rainbow trout feed additive of promotion growth, preparation method and applications |
| CN109258930A (en) * | 2018-11-01 | 2019-01-25 | 青岛农业大学 | A kind of one-step fermentation of dregs of beans, the fermented bean dregs of this method preparation and application |
| WO2023055850A1 (en) * | 2021-09-29 | 2023-04-06 | Dupont Nutrition Biosciences Aps | Optimized water line delivery of microbes |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104805031A (en) * | 2014-09-17 | 2015-07-29 | 德强生物股份有限公司 | Bacillus subtilis and preparation method and application thereof |
| CN104611257A (en) * | 2014-12-25 | 2015-05-13 | 浙江惠嘉生物科技有限公司 | Preparation method of bacillus licheniformis preparations for drinking water of livestock and poultry |
| CN107236691A (en) * | 2017-06-28 | 2017-10-10 | 四川农业大学 | A kind of beaver rabbit source bacillus subtilis and its application in growth of Rex rabbits performance is improved |
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| CN108125067A (en) * | 2017-12-01 | 2018-06-08 | 杭州柏普泰生物科技有限公司 | A kind of disease preventing and treating and the rainbow trout feed additive of promotion growth, preparation method and applications |
| CN109258930A (en) * | 2018-11-01 | 2019-01-25 | 青岛农业大学 | A kind of one-step fermentation of dregs of beans, the fermented bean dregs of this method preparation and application |
| WO2023055850A1 (en) * | 2021-09-29 | 2023-04-06 | Dupont Nutrition Biosciences Aps | Optimized water line delivery of microbes |
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Application publication date: 20120912 |