CN102787086A - Culture medium for culturing bacillus subtilis for feedstuffs - Google Patents
Culture medium for culturing bacillus subtilis for feedstuffs Download PDFInfo
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- CN102787086A CN102787086A CN2012102631911A CN201210263191A CN102787086A CN 102787086 A CN102787086 A CN 102787086A CN 2012102631911 A CN2012102631911 A CN 2012102631911A CN 201210263191 A CN201210263191 A CN 201210263191A CN 102787086 A CN102787086 A CN 102787086A
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- fish meal
- culture medium
- subtilis
- bacillus subtilis
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Abstract
The invention relates to a culture medium for culturing bacillus subtilis for feedstuffs. The culture medium comprises the following components in percentage by weight: 2%-5% of fish meal waste water concentrated solution, 1% of glucose, 0.5% of sodium chloride, 0.3% of monopotassium phosphate, 0.15% of magnesium sulfate and the balance of water. According to the culture medium for culturing the bacillus subtilis for the feedstuffs disclosed by the invention, the fish meal waste water concentrated solution is used as a new raw material for producing the bacillus subtilis BL-8, and the culturing cost of the bacillus subtilis can be greatly reduced; the number of zymocytes of the bacillus is improved, and spore production rate of the bacillus subtilis is improved; and a new way is developed for the use of the fish meal waste water, and the problems that the environment is polluted by the fish meal waste water which is a by-product in the fish meal production and the fish meal waste water is difficult to treat are solved.
Description
Technical field:
The present invention relates to the bacillus culture medium that a kind of feed uses.
Background technology:
Subtilis LB-8 draws from Chinese microbial preservation center; Deposit number is: CGMCC No.1.210; Be the genus bacillus that can be used for feed, the ability of its synthetic AMS is strong, uses it for herding, poultry, aquatic products, special animals and pet and cultures; Can prevent that animal digestion is bad, improve the utilization ratio of feed; Can also prevent functions such as intestines and stomach disease such as animal diarrhoea, diarrhea; Subtilis LB-8 exists with endogenous gemma form, is a kind of stalwartness and be in the viable cell of dormant state, strong stress resistance; Storage time is long; No matter it is at granulated feed or in mash feed, all more stable, can get into animal intestinal smoothly and become also a large amount of propagation alive.Feedstuff industry has been applied for many years, and effect is very good.The cultivation feed that uses at present is with the substratum of subtilis, and its each component and weight percent are following: glucose 1%, and peptone 1.5%, sodium-chlor 0.5%, Carnis Bovis seu Bubali cream 0.05%, potassium primary phosphate 0.3%, sal epsom 0.15%, remaining amount is water.Its weak point is that production cost is high.
Summary of the invention
The object of the present invention is to provide a kind of gemma rate height and low a kind of substratum of cultivating feed with subtilis of cost of producing.The present invention cultivates feed with the fish meal wastewater liquid concentrator as main nutrient matter to use subtilis LB-8.Each component of the present invention and weight percent are following: fish meal wastewater liquid concentrator 2-5%, and glucose 1%, sodium-chlor 0.5%, potassium primary phosphate 0.3%, sal epsom 0.15%, remaining amount is water.Wherein, the fish meal wastewater liquid concentrator is during fish meal is produced, and the liquid that its fresh fish squeezes out after boiling makes moisturely at 60% light brown liquid through concentrating, and except that having fishy smell, does not have other peculiar smell, protein contnt 25-30% (weight), pH value 5.0-7.0.
The preparation method of substratum of the present invention: the ratio in culture medium prescription of the present invention takes by weighing glucose, sodium-chlor, potassium primary phosphate and sal epsom, puts into beaker, gets the fish meal wastewater liquid concentrator in the ratio of prescription again; In beaker, add the water that is less than specified quantity then; Stir, the heating, treat that above-mentioned each component is dissolved fully after; Transfer pH to 7.0 with 1-5mol/LNaOH, make up water is to the quantity of regulation.With prepared culture medium packing as required, seal film and seal with ventilative then, 121.3 ℃ of high pressure steam sterilizations 30 minutes.
The present invention compared with prior art has following advantage:
1) the fermentative prodn genus bacillus was many in the past is raw materials for production with dregs of beans, starch, and production cost is high, and the present invention as a kind of new raw material production subtilis BL-8, can reduce the fish meal wastewater liquid concentrator greatly subtilis and cultivate cost;
2) improve fermentation of bacillus bacterium number, improve the product gemma rate of subtilis;
3) open up new way for the utilization of fish meal wastewater, solve sub product fish meal wastewater contaminate environment, unmanageable problem that fish meal is produced.
Description of drawings
Fig. 1 is 24 hours viable count detection case figure of control group fermentation.
Fig. 2 is the present invention 24 hours viable count detection case figure that ferment.
Fig. 3 is 24 hours growing state micro-imaging figure (1000 *) of control group fermentation of bacillus.
Fig. 4 is 24 hours growing state micro-imaging figure (1000 *) of fermentation of bacillus of the present invention.
Fig. 5 is 44 hours genus bacillus growing state micro-imaging figure (1000 *) of control group fermentation.
Fig. 6 is the present invention 44 hours genus bacillus growing state micro-imaging figure (1000 *) that ferment.
Fig. 7 is 48 hours genus bacillus growing state micro-imaging figure (1000 *) of control group fermentation.
Fig. 8 is the present invention 48 hours genus bacillus growing state micro-imaging figure (1000 *) that ferment.
Fig. 9 is 60 hours genus bacillus growing state micro-imaging figure (1000 *) of control group fermentation.
Figure 10 is the present invention 60 hours genus bacillus growing state micro-imaging figure (1000 *) that ferment.
Embodiment
Example 1
Accurately take by weighing glucose 10 grams, sodium-chlor 5 grams, potassium primary phosphate 3 and sal epsom 1.5 grams, put into beaker, take by weighing fish meal wastewater liquid concentrator 20 grams again; In beaker, add 800 gram water then; Stir, the heating, treat that above-mentioned each component is dissolved fully after; Transfer pH to 7.0 with 5mol/LNaOH, make up water to 1000 milliliter.The prepared culture medium branch is filled to 250 milliliters triangular flask, and 60 milliliters of every bottled amounts are sealed film and are sealed with ventilative, 121.3 ℃ of high pressure steam sterilizations 30 minutes.
Example 2
Accurately take by weighing glucose 10 grams, sodium-chlor 5 grams, potassium primary phosphate 3 and sal epsom 1.5 grams, put into beaker, take by weighing fish meal wastewater liquid concentrator 50 grams again; In beaker, add 800 gram water then; Stir, the heating, treat that above-mentioned each component is dissolved fully after; Transfer pH to 7.0 with 1mol/LNaOH, make up water to 1000 milliliter.The prepared culture medium branch is filled to 250 milliliters triangular flask, and 60 milliliters of every bottled amounts are sealed film and are sealed with ventilative, 121.3 ℃ of high pressure steam sterilizations 30 minutes.
Test-results
1, produces gemma rate simultaneous test
The subtilis LB-8 of the same terms is transferred respectively in prior art substratum and substratum of the present invention, and 37 ℃ of shake flask fermentation liquid are cultivated.Wherein, the number of viable of prior art culture medium culturing 24h is 4.41 * 10
9Cfu/mL fermented 48 hours, produced gemma rate 85%; The number of viable of culture medium culturing 24h of the present invention is 1.77 * 10
11Cfu/mL fermented 48 hours, produced gemma rate 94%, and culture effect obviously is superior to the prior art beef-protein medium.
2, fermentation test
1. liquid seeds preparation: preparation beef extract-peptone liquid nutrient medium (pH value 7.2-7.5), be sub-packed in 18mm * 180mm glass test tube, every pipe loading amount 10ml, 121.3 ℃ of high pressure steam sterilizations 20 minutes cool off subsequent use.Get one (inoculation) and encircle the subtilis that purifying is good inclined-plane is preserved, be inoculated in the liquid nutrient medium, in 37 ℃, 200r/min shaking table shaking culture 12h is seed liquor.
2. liquid fermentation and culture: control group (prior art) substratum and substratum of the present invention are inserted the seed liquor that has prepared by 1% inoculum size respectively, and 37 ℃, the 200r/min shaking table is cultivated, and when cultivating 24h, detects the fermentation viable count.
Get 1 mL nutrient solution respectively to the sterilized water of 9ml from above-mentioned control group and of the present invention group, dilute by 1:10 series, 1:1 * 10
1, 1:1 * 10
2, 1:1 * 10
3, 1:1 * 10
4, 1:1 * 10
5, 1:1 * 10
6, 1:1 * 10
7, 1:1 * 10
8, 1:1 * 10
9Use the sterilized water diluent, get 1:1 * 10
8, 1:1 * 10
9Each 0.1 mL of diluent coats the bacterium number and detects on the culture medium flat plate, cultivates and observe the colony count that grows for 37 ℃.Fig. 1 is 24 hours colony count detection case figure (1:1 * 10 of control group fermentation
8Diluent), Fig. 2 is the present invention 24 hours colony count detection case figure (1:1 * 10 of fermenting
8Diluent), can find out that from two width of cloth figure contrast under the same terms, the colony count of culture medium culturing of the present invention is obviously more than control group.And be 4.41 * 10 through detecting, calculating control group subtilis viable count
9Cfu/mL, of the present invention group of subtilis number of viable is 1.77 * 10
11Cfu/mL is higher than control group far away.
Subtilis viable bacteria in above-mentioned control group and of the present invention group is continued to cultivate; And respectively at 24h, 44h, 48h and 60h micro-imaging; Be Fig. 3 and Fig. 4, Fig. 5 and Fig. 6, Fig. 7 and Fig. 8, Fig. 9 and Figure 10. observe control group and of the present invention group of bacillus subtilis spore production, control group producing bacillus subtilis gemma rate is starkly lower than of the present invention group of producing bacillus subtilis gemma rate, and through detecting; Fermentation culture 48h; Control group producing bacillus subtilis gemma rate is 85%, and of the present invention group product gemma rate 94% is higher than control group.
Claims (3)
1. substratum of cultivating feed with subtilis, it is characterized in that: its each component and weight percent are following: fish meal wastewater liquid concentrator 2-5%, glucose 1%, sodium-chlor 0.5%, potassium primary phosphate 0.3%, sal epsom 0.15%, remaining amount is water.
2. cultivation feed according to claim 1 is characterized in that with the substratum of subtilis: the fish meal wastewater liquid concentrator is during fish meal is produced, and the liquid that its fresh fish squeezes out after boiling makes weight in wet base compare the light brown liquid 60% through concentrating.
3. the cultivation feed of claim 1 is characterized in that with the preparation method of the substratum of subtilis: the ratio in culture medium prescription of the present invention takes by weighing glucose, sodium-chlor, potassium primary phosphate and sal epsom, puts into beaker; Get the fish meal wastewater liquid concentrator in the ratio of prescription again, in beaker, add the water that is less than specified quantity then, stir; Heating, treat that above-mentioned each component is dissolved fully after, transfer pH to 7.0 with 1-5mol/LNaOH; Make up water is to the quantity of regulation; With prepared culture medium packing as required, seal film and seal with ventilative then, 121.3 ℃ of high pressure steam sterilizations 30 minutes.
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| Application Number | Priority Date | Filing Date | Title |
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| CN2012102631911A CN102787086A (en) | 2012-07-27 | 2012-07-27 | Culture medium for culturing bacillus subtilis for feedstuffs |
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| CN2012102631911A CN102787086A (en) | 2012-07-27 | 2012-07-27 | Culture medium for culturing bacillus subtilis for feedstuffs |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104381605A (en) * | 2014-11-06 | 2015-03-04 | 中山市巴斯德农业科技有限公司 | Microbial feed additive and preparation method thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101348776A (en) * | 2008-07-31 | 2009-01-21 | 东北师范大学 | Transgenic lactobacillus secreting acidic cellulase, preparation and use thereof |
-
2012
- 2012-07-27 CN CN2012102631911A patent/CN102787086A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101348776A (en) * | 2008-07-31 | 2009-01-21 | 东北师范大学 | Transgenic lactobacillus secreting acidic cellulase, preparation and use thereof |
Non-Patent Citations (2)
| Title |
|---|
| 张东升等: "利用鱼粉生产中的废水培养苏云金芽胞杆菌的可行性研究", 《大连水产学院学报》 * |
| 朱碧英等: "鱼粉废弃榨液及其浓缩液成分分析研究", 《饲料研究》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104381605A (en) * | 2014-11-06 | 2015-03-04 | 中山市巴斯德农业科技有限公司 | Microbial feed additive and preparation method thereof |
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Application publication date: 20121121 |