CN103805547B - A kind of plant lactobacillus LP-12 and the application in the efficient animal intestinal conditioning agent of preparation thereof - Google Patents
A kind of plant lactobacillus LP-12 and the application in the efficient animal intestinal conditioning agent of preparation thereof Download PDFInfo
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- CN103805547B CN103805547B CN201410081341.6A CN201410081341A CN103805547B CN 103805547 B CN103805547 B CN 103805547B CN 201410081341 A CN201410081341 A CN 201410081341A CN 103805547 B CN103805547 B CN 103805547B
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Abstract
The invention discloses a kind of plant lactobacillus LP-12 and the application in the efficient animal intestinal conditioning agent of preparation thereof.Does is the culture presevation number of this plant lactobacillus LP-12: CCTCC? M? 2012401, it has the function regulating microbial population of animal intestinal tract, strengthen animal immunizing power, can be used for preparing efficient animal intestinal conditioning agent.The invention also discloses a kind of efficient animal intestinal conditioning agent, containing bacterium powder, protein raw materials and the reinforcing stimulus factor in this efficient animal intestinal conditioning agent, described bacterium powder is the bacterium powder of plant lactobacillus LP-12 of the present invention.Efficient animal intestinal conditioning agent of the present invention, by the bacterium powder of plant lactobacillus LP-12 of the present invention, with bean cake powder with strengthen stimulating factor and carry out compatibility in proportion, there is very strong enteron aisle regulating power, medicine usage quantity can be reduced after using, improve animal immunizing power, strengthen stress ability, to the aquaculture volume increase of promotion China, promote that green cultivation is significant.
Description
Technical field
The present invention relates to a kind of can be used for producing efficient animal intestinal conditioning agent lactobacillus plantarum and the efficient animal intestinal conditioning agent produced of this bacterial strain a kind of.
Technical background
For a long time, further developing of livestock and poultry intensive farm increases with its feeding density, and the generation of various livestock and poultry is more and more frequent, and the use of probiotic bacterium can greatly reduce antibiotic use.Microbiotic has control and disease therapy, improves the functions such as the price of deed, and antibiotic use improves fanning economics on the whole, for promoting that the development of livestock and poultry breeding industry has played positive effect.But due to the shortage with scientific knowledge of ordering about of economic interests, the phenomenon ubiquity of Drug abuse in aquaculture.Abuse of antibiotics can cause microbiotic remaining in animal food, affects the health of the mankind; And after animal uses microbiotic, medicine is got rid of with the movement such as excrement, urine with the form of original shape or metabolite, residues in environment.Along with the enhancing of countries in the world environmental consciousness, people more and more pay close attention to veterinary drug accumulation in the environment, transfer, conversion and the impact on various biology and human health.And probiotic bacterium is extensively present in the enteron aisle of humans and animals, wherein topmost is milk-acid bacteria, and this is the bacterium that a group is quite numerous and jumbled, at least can be divided into 18 genus at present, have kind more than 200.Except only a few, wherein the overwhelming majority is all that animal body is requisite and have the flora of important physiological function.In gi tract microflora group and the microecosystem that forms between animal and environment, dominant population in microflora plays conclusive effect to whole microflora, once lose dominant population, micro ecology of gastrointestinal tract system balancing can be caused to lack of proper care, original dominant population changes, and animal occurs that production performance declines or disease symptoms.Show according to research data recent years: the probiotic bacterium based on milk-acid bacteria has various health-care as the profitable strain of settling down in enteron aisle: 1) colonization resistance and ecological occupy-place.Microecological balance and intestinal tube function can be maintained.The Growth and reproduction of unwanted bacteria can be suppressed, maintain the normal function of the eubiosis and enteron aisle in intestines, can also intestines peristalsis be stimulated, increase ight soil water content, relax bowel.2) Membranous flora barrier action.Be added with in animal-feed beneficial microorganism can the attached collection of competitive inhibition pathogenic agent on gastrointestinal tract wall, play shielding effect, stop the field planting of pathogenic bacterium and invasion, the body that watches for animals is from infection.3) immunologic function is improved.Reduce animal body to fall ill rate, reduce the use of medicine, reduce mortality ratio.4) promoting digestion.Probiotic bacterium can produce various digestive ferment in animal body, improves food conversion ratio.
How cultivating under few use does not even use antibiotic condition, probiotic bacterium will become one of the most useful weapon.And screen a kind of good stability, enteron aisle regulating effect well efficient microorganism, be the key of head it off, and with a kind of quick-acting, stable, cheap animal intestinal conditioning agent of its production, China's aquaculture had great importance.
Summary of the invention
For the deficiencies in the prior art, technical problem solved by the invention is to provide a kind of plant lactobacillus LP-12 with good enteron aisle regulating effect, good stability, to overcome the shortcoming of existing microbial preparation in stability, vigor and enteron aisle regulating effect.
Technical problem solved by the invention is also to provide the application of above-mentioned plant lactobacillus LP-12 in the efficient animal intestinal conditioning agent of preparation.
Technical problem solved by the invention is also to provide a kind of efficient animal intestinal conditioning agent, and it has quick-acting, stable, cheap advantage, for improving animal-feed rate of utilization, strengthening animal immunizing power.
To achieve these goals, a kind of plant lactobacillus LP-12 provided by the invention (
lactobacillusplantarumlP-12), this bacterial strain is on October 14th, 2012 in China typical culture collection center (address: China, Wuhan, Wuhan University) preservation, and deposit number is CCTCCNO:M2012401, and this bacterial strain has antibacterial and efficient enteron aisle regulating power.
Above-mentioned bacterial classification has following character:
1, Morphology and physiology feature:
Individual cells is 0.9 ~ 1.2 × 3 ~ 8 microns, exists, gram-positive microorganism with single, paired or the arrangement of short chain form, without pod membrane, and atrichia, stiff.Mycoderm is formed during quiescent culture in liquid medium within, facultative aerobic.In LBS nutrient agar, growth is in canescence, can form opaque, circular, smooth, small fine and closely woven bacterium colony.
2, the substratum adopted:
Solid slope is separated and counting substratum: glucose 2%, yeast extract paste 1%, sodium acetate 0.5%, Tween-800.2%, ammonium citrate 0.2%, K
2hPO
40.2%, MgSO
47H
2o0.06%, MnSO
4h
2o0.02%, agar 2%, pH6.5,115 DEG C, sterilizing 20min.
Liquid seed culture medium: glucose 5%, peptone 0.5%, yeast extract paste 1%, peptone 1%, extractum carnis 1%, K
2hPO
40.2%, MgSO
47H
2o0.06%, MnSO
4h
2o0.02%, pH value 6.5,115 DEG C of sterilizing 20min.
Fermentation production medium: glucose 2 ~ 5%, peptone 0.2 ~ 2%, yeast extract paste 0.3 ~ 2%, calcium carbonate 1 ~ 12%, K
2hPO
40.1 ~ 0.8%, MgSO
47H
2o0.02 ~ 0.1%, MnSO
4h
2o0.01 ~ 0.05%, pH value 5.5 ~ 7.5,115 DEG C of sterilizing 20min.
3, culture condition:
Culture temperature: 35 DEG C ~ 40 DEG C; Cultivate pH6.5
4, authentication method:
Bacterial strain sample is pressed respectively SDS method and extracts STb gene, after pcr amplification, ITS sequence is measured, use Blast instrument sequencing result and ncbi database to be contrasted, determine bacterial strain be plant lactobacillus (
lactobacillusplantarum) Identities=96%, Gaps=1%.
5, the production technique of bacterial strain of the present invention, at least comprises the following steps: 1) cultivate bacterial strain; 2) make substratum and ferment; 3) carrier is added; 4) dry
Described production technique also comprises: 1) bacterial strain seed culture; 2) fermentation processes, pH controls; 3) add carrier, promote bacterial strain embedding; 4) spraying dry.
In addition, present invention also offers a kind of efficient animal intestinal conditioning agent, containing bacterium powder, protein raw materials and the reinforcing stimulus factor in this efficient animal intestinal conditioning agent, described bacterium powder is the bacterium powder of above-mentioned plant lactobacillus LP-12.
Specifically, the bacterium powder for efficient animal intestinal conditioning agent of the present invention is prepared from as follows:
1, from inclined plane inoculating to shake-flask seed substratum, cultivate after 18 ~ 24 hours with in the inoculum size access fermentor tank of 0.2% ~ 1% for 36 DEG C ~ 38 DEG C.Fermentation culture conditions is: cultivate 48 ~ 56 hours for 37 DEG C.
2, rear interpolation absorption carrier has been cultivated: sucrose 1% ~ 5%, dextrin 2% ~ 8%, skim-milk 2% ~ 4%.
3, spray dried drying prescription is used to carry out drying, obtained bacterium powder.
4, in bacterium powder, the total count of plant lactobacillus LP-12 is 180 × 10
9~ 360 × 10
9cfu/g.
Wherein, by weight percentage, described bacterium powder accounts for 0.2% ~ 2% of described efficient animal intestinal conditioning agent.
Preferably, described protein raw materials is bean cake powder.
Preferably, containing accounting for the following composition of this efficient animal intestinal conditioning agent weight percent in the described reinforcing stimulus factor: magnesium sulfate 0.15% ~ 0.2%, manganous sulfate 0.8% ~ 0.21%, Sodium phosphate dibasic 0.1% ~ 0.3%, ammonium citrate 0.1% ~ 0.4%, peptone 0.5% ~ 1%, glucose 1% ~ 3%.
In the configuration and constituent optimization process of the efficient animal intestinal conditioning agent of the present invention, the bacterium powder of obtained above-mentioned plant lactobacillus LP-12 is mixed with protein raw materials, bacterium powder occupies 3 ~ 5% (weight ratios) of protein raw materials, again bacterium powder after mixing is fully mixed with the reinforcing stimulus factor, last granulation.
The present invention devises reinforcing stimulus factor recipe ingredient, to strengthen the effect Tanaka.The group of the reinforcing stimulus factor wants composition to be: magnesium sulfate, manganous sulfate, Sodium phosphate dibasic, ammonium citrate, peptone, glucose.
The composition of the reinforcing stimulus factor after being optimized by various experiment is: magnesium sulfate 0.15% ~ 0.2%, manganous sulfate 0.21% ~ 0.80%, Sodium phosphate dibasic 0.1% ~ 0.3%, ammonium citrate 0.1% ~ 0.4%, peptone 0.5% ~ 1%, glucose 1% ~ 3%, wherein each degree refers to and accounts for this efficient animal intestinal conditioning agent weight percent.
The efficient animal intestinal conditioning agent adopting bacterial classification of the present invention and production method to produce, can directly be mixed in feed, can improve animal intestinal health, promoting digestion, raising immunizing power, improves that animal disease resistant is poor, degradation symptom under yield and quality.And low production cost, protein raw materials is simply extensive.Adopt bacterial classification of the present invention, good stability, be beneficial to preservation, long quality-guarantee period.
By illustrating in greater detail the present invention by following examples, be that following examples are only illustrative with it may be noted that, the present invention is not by the restriction of these embodiments.
embodiment:
Embodiment one: the production of bacterium powder.
1, from inclined plane inoculating to liquid seed culture medium, cultivate 24 hours for 37 DEG C.
Wherein solid slant culture base is: glucose 2%, yeast extract paste 1%, sodium acetate 0.5%, Tween-800.2%, ammonium citrate 0.2%, K
2hPO
40.2%, MgSO
47H
2o0.06%, MnSO
4h
2o0.02%, agar 2%, pH6.5,115 DEG C, sterilizing 20min.
Liquid seed culture medium is: glucose 5%, peptone 0.5%, yeast extract paste 1%, peptone 1%, extractum carnis 1%, K
2hPO
40.2%, MgSO
47H
2o0.06%, MnSO
4h
2o0.02%, pH value 6.5,115 DEG C of sterilizing 20min.
2, seed culture complete after with 0.2% inoculum size access fermentor tank in.
Fermention medium is: glucose 3%, peptone 1%, yeast extract paste 1%, calcium carbonate 6%, K
2hPO
40.4%, MgSO
47H
2o0.06%, MnSO
4h
2o0.03%, pH value 6.5.
Fermentation culture conditions is: cultivate 50 hours for 37 DEG C, rotating speed 250rpm, Anaerobic culturel
3, rear interpolation absorption carrier has been cultivated: 1%CaCO
3, 2% wheat bran and flocculating aids: 2% perlite, 1% zeolite powder.Use plate-and-frame filter press filters.
4, in filter residue, add sucrose 3%, dextrin 5%, skim-milk 4%, carry out spraying dry, air outlet temperature is 120 DEG C.
5, collect bacterium powder, the total count detecting plant lactobacillus LP-12 in bacterium powder is 234 × 10
9cfu/g.
Embodiment two: the production of efficient animal intestinal conditioning agent.
1, select bean cake powder, add the bacterium powder of 1% in mass ratio, fully mix.
2, the reinforcing stimulus factor is added by after mixing.
3, the composition of the reinforcing stimulus factor is: magnesium sulfate 0.16%, manganous sulfate 0.1%, Sodium phosphate dibasic 0.2%, ammonium citrate 0.3%, peptone 0.6%, glucose 2%.
4, granulation after mixing.
5, fully the rear shady and cool dry place of mixing preserves.
Be 1.9 × 10 through checking the viable count of this efficient animal intestinal conditioning agent
9cfu/g, total protein content is 36%.
Embodiment three: the production of efficient animal intestinal conditioning agent.
1, select bean cake powder, add the bacterium powder of 0.3% in mass ratio, fully mix.
2, the reinforcing stimulus factor is added by after mixing.
3, the composition of the reinforcing stimulus factor is: magnesium sulfate 0.15%, manganous sulfate 0.8%, Sodium phosphate dibasic 0.1%, ammonium citrate 0.1%, peptone 0.5%, glucose 3%.
4, granulation after mixing.
5, fully the rear shady and cool dry place of mixing preserves.
Be 0.63 × 10 through checking the viable count of this efficient animal intestinal conditioning agent
9cfu/g, total protein content is 38%.
Execute example four: the production of efficient animal intestinal conditioning agent.
1, select bean cake powder, add the bacterium powder of 2% in mass ratio, fully mix.
2, the reinforcing stimulus factor is added by after mixing.
3, the composition of the reinforcing stimulus factor is: magnesium sulfate 0.2%, manganous sulfate 0.21%, Sodium phosphate dibasic 0.3%, ammonium citrate 0.4%, peptone 1%, glucose 1%.
4, granulation after mixing.
5, fully the rear shady and cool dry place of mixing preserves.
Be 4.3 × 10 through checking the viable count of this efficient animal intestinal conditioning agent
9cfu/g, total protein content is 34%.
Embodiment five: application test.
Laboratory animal: piglet.
Test sample: use normal raising to do blank in an experiment.
The total class of animal: landrace.
Test-results:
| Project | To feed quantity | To feed number of days | Initial pig heavy (kg) | Terminate pig heavy (kg) | Day weight gain (g) | Total feed consumption (kg) | Day feed consumption (g) | Feed-weight ratio |
| Experimental group 1 | 264 | 20 | 16.6 | 27.9 | 565 | 23.8 | 1190 | 2.11 |
| Experimental group 2 | 83 | 20 | 15.7 | 26.8 | 557 | 24.1 | 1208 | 2.17 |
| Experimental group 3 | 138 | 20 | 16.2 | 28.2 | 599 | 25.6 | 1281 | 2.14 |
| Control group | 273 | 20 | 16.7 | 27.3 | 530 | 23.7 | 1185 | 2.24 |
Note: the highest day weight gain can improve 6.6%, feed-weight ratio reduces by 5.7%.
Find obviously to improve the hair color of animal and reduced the generation of breeding environment stink, having improved breeding environment, cultivated animals fur light.
Embodiment four: application test.
Laboratory animal: broiler chicken.
Test sample: use normal raising to do blank in an experiment.
Test method:
Find obviously to improve the hair color of animal and reduced the generation of breeding environment stink, having improved breeding environment.
The above is the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the premise without departing from the principles of the invention; can also make some improvements and modifications, these improvements and modifications are also considered as protection scope of the present invention.
Claims (4)
1. an efficient animal intestinal conditioning agent, it is characterized in that: containing bacterium powder, protein raw materials and the reinforcing stimulus factor in this efficient animal intestinal conditioning agent, described bacterium powder is the bacterium powder of plant lactobacillus LP-12, the deposit number of plant lactobacillus LP-12 is CCTCCM2012401, has efficient enteron aisle regulating power;
Described bacterium powder is prepared from accordance with the following steps:
(1), from inclined plane inoculating to shake-flask seed substratum, cultivate after 18 ~ 24 hours with in the inoculum size access fermentor tank of 0.2% ~ 1% for 36 DEG C ~ 38 DEG C;
Fermentation culture conditions is: cultivate 48 ~ 56 hours for 37 DEG C;
(2), rear interpolation absorption carrier has been cultivated: sucrose 1% ~ 5%, dextrin 2% ~ 8%, skim-milk 2% ~ 4%;
(3), use spray dried drying prescription to carry out drying, obtained bacterium powder, the total count content of plant lactobacillus LP-12 described in bacterium powder is 180 × 10
9~ 360 × 10
9cfu/g.
2. efficient animal intestinal conditioning agent according to claim 1, is characterized in that: by weight percentage, and described bacterium powder accounts for 0.2% ~ 2% of described efficient animal intestinal conditioning agent.
3. efficient animal intestinal conditioning agent according to claim 1, is characterized in that: described protein raw materials is bean cake powder.
4. efficient animal intestinal conditioning agent according to claim 1, it is characterized in that, containing accounting for the following composition of this efficient animal intestinal conditioning agent weight percent in the described reinforcing stimulus factor: magnesium sulfate 0.15% ~ 0.2%, manganous sulfate 0.8% ~ 0.21%, Sodium phosphate dibasic 0.1% ~ 0.3%, ammonium citrate 0.1% ~ 0.4%, peptone 0.5% ~ 1%, glucose 1% ~ 3%.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106635894A (en) * | 2016-11-29 | 2017-05-10 | 内蒙古和美科盛生物技术有限公司 | Preservative bacterium powder for uncooked feed solid fermentation and preparation process of preservative bacterium powder |
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| CN112501063B (en) * | 2020-12-03 | 2022-03-08 | 兰州大学 | Lactobacillus plantarum LZU-J-TSL6 capable of highly producing GABA and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1644681A (en) * | 2004-12-17 | 2005-07-27 | 上海交大昂立股份有限公司 | Plant lactobacillaceae and use thereof |
| CN1985854A (en) * | 2006-12-22 | 2007-06-27 | 上海交大昂立股份有限公司 | Plant lactobacillus composition and its application in strengthening intestinal immunobarrier |
| CN101955549A (en) * | 2010-09-28 | 2011-01-26 | 中国农业大学 | Large-headed atractylodes rhizome polysaccharide and preparation method and application thereof |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1644681A (en) * | 2004-12-17 | 2005-07-27 | 上海交大昂立股份有限公司 | Plant lactobacillaceae and use thereof |
| CN1985854A (en) * | 2006-12-22 | 2007-06-27 | 上海交大昂立股份有限公司 | Plant lactobacillus composition and its application in strengthening intestinal immunobarrier |
| CN101955549A (en) * | 2010-09-28 | 2011-01-26 | 中国农业大学 | Large-headed atractylodes rhizome polysaccharide and preparation method and application thereof |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106635894A (en) * | 2016-11-29 | 2017-05-10 | 内蒙古和美科盛生物技术有限公司 | Preservative bacterium powder for uncooked feed solid fermentation and preparation process of preservative bacterium powder |
| CN106635894B (en) * | 2016-11-29 | 2019-06-07 | 内蒙古和美科盛生物技术有限公司 | A kind of fresh-keeping bacterium powder and its preparation process for raw material solid fermentation |
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