CN111919964A - Forsythia suspensa leaf-based microecological preparation and preparation method thereof - Google Patents
Forsythia suspensa leaf-based microecological preparation and preparation method thereof Download PDFInfo
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- 241000576429 Forsythia suspensa Species 0.000 title claims description 6
- 241000555712 Forsythia Species 0.000 claims abstract description 70
- 239000000843 powder Substances 0.000 claims abstract description 22
- 238000000855 fermentation Methods 0.000 claims abstract description 20
- 244000144972 livestock Species 0.000 claims abstract description 12
- 244000144977 poultry Species 0.000 claims abstract description 11
- 244000063299 Bacillus subtilis Species 0.000 claims abstract description 10
- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 10
- 239000003674 animal food additive Substances 0.000 claims abstract description 10
- 235000013960 Lactobacillus bulgaricus Nutrition 0.000 claims abstract description 9
- 229940004208 lactobacillus bulgaricus Drugs 0.000 claims abstract description 9
- 241000235646 Cyberlindnera jadinii Species 0.000 claims abstract description 7
- 238000010564 aerobic fermentation Methods 0.000 claims abstract description 7
- 239000002994 raw material Substances 0.000 claims abstract description 5
- 241000186672 Lactobacillus delbrueckii subsp. bulgaricus Species 0.000 claims abstract 3
- 238000001035 drying Methods 0.000 claims description 9
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- 208000027312 Bursal disease Diseases 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
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- DTOUWTJYUCZJQD-QJDQKFITSA-N Forsythiaside Natural products C[C@@H]1O[C@H](OC[C@H]2O[C@@H](OCCc3ccc(O)c(O)c3)[C@H](O)[C@@H](O)[C@@H]2OC(=O)C=Cc4ccc(O)c(O)c4)[C@H](O)[C@H](O)[C@H]1O DTOUWTJYUCZJQD-QJDQKFITSA-N 0.000 description 1
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- 235000013330 chicken meat Nutrition 0.000 description 1
- CYDMQBQPVICBEU-UHFFFAOYSA-N chlorotetracycline Natural products C1=CC(Cl)=C2C(O)(C)C3CC4C(N(C)C)C(O)=C(C(N)=O)C(=O)C4(O)C(O)=C3C(=O)C2=C1O CYDMQBQPVICBEU-UHFFFAOYSA-N 0.000 description 1
- 229960004475 chlortetracycline Drugs 0.000 description 1
- CYDMQBQPVICBEU-XRNKAMNCSA-N chlortetracycline Chemical compound C1=CC(Cl)=C2[C@](O)(C)[C@H]3C[C@H]4[C@H](N(C)C)C(O)=C(C(N)=O)C(=O)[C@@]4(O)C(O)=C3C(=O)C2=C1O CYDMQBQPVICBEU-XRNKAMNCSA-N 0.000 description 1
- 235000019365 chlortetracycline Nutrition 0.000 description 1
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- 235000012343 cottonseed oil Nutrition 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 1
- 235000019797 dipotassium phosphate Nutrition 0.000 description 1
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- 230000007774 longterm Effects 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
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- 229940099596 manganese sulfate Drugs 0.000 description 1
- 235000007079 manganese sulphate Nutrition 0.000 description 1
- 239000011702 manganese sulphate Substances 0.000 description 1
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
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- 239000011574 phosphorus Substances 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 229920000136 polysorbate Polymers 0.000 description 1
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 1
- 235000013613 poultry product Nutrition 0.000 description 1
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- 229940126680 traditional chinese medicines Drugs 0.000 description 1
- YWYZEGXAUVWDED-UHFFFAOYSA-N triammonium citrate Chemical compound [NH4+].[NH4+].[NH4+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O YWYZEGXAUVWDED-UHFFFAOYSA-N 0.000 description 1
- 239000001393 triammonium citrate Substances 0.000 description 1
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/70—Feeding-stuffs specially adapted for particular animals for birds
- A23K50/75—Feeding-stuffs specially adapted for particular animals for birds for poultry
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Polymers & Plastics (AREA)
- Animal Husbandry (AREA)
- Biotechnology (AREA)
- Food Science & Technology (AREA)
- Zoology (AREA)
- Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Physiology (AREA)
- Birds (AREA)
- Microbiology (AREA)
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Abstract
The invention relates to a micro-ecological preparation based on forsythia suspense leaves, which is prepared by taking dry forsythia suspense leaf powder as a raw material, inoculating bacillus subtilis and candida utilis to perform aerobic fermentation, and then inoculating lactobacillus bulgaricus of Germany to perform anaerobic fermentation. The microecological preparation is used as a livestock and poultry feed additive, is directly added into livestock and poultry feed to feed broiler chickens, can improve the utilization rate of broiler chicken feed, effectively improve the production performance of broiler chickens, reduce the diarrhea rate and the death rate of broiler chickens, obviously improve the thymus index and the bursa of fabricius index of broiler chickens, increase the IgA and IgM contents in serum, enhance the immunologic function of broiler chickens and reduce the use of antibiotics.
Description
Technical Field
The invention belongs to the technical field of medicinal plant application, and relates to a preparation method of a forsythia suspense leaf microecological preparation.
Background
In the production of animal husbandry, the problems of drug residue and gradually enhanced drug resistance of pathogenic bacteria in livestock and poultry bodies are increasingly prominent due to long-term or overdose use of feed antibiotics. The Chinese herbal medicine has the characteristics of low toxicity, low residue, no drug resistance, multiple functions and the like, and has good effects of enhancing the immunity of livestock and poultry organisms, improving protein metabolism and calcium and phosphorus metabolism, improving the production performance, preventing and treating diseases and increasing the fragrance of feed. It has become a research hotspot as an alternative to antibiotics.
Fructus forsythiae is a plant of the Oleaceae family, is a traditional Chinese herbal medicine, has the effects of clearing heat and removing toxicity, and reducing swelling and dissipating stagnation, and is mainly used for treating diseases such as tepidity, erysipelas, macula, carbuncle and swelling and the like. Modern scientific research proves that the forsythia has excellent antibacterial and antiviral activity. Fructus forsythiae is traditionally used as a medicine by fruits. In a long period of time, the development of wild forsythia is unbalanced, the green-catching phenomenon of the production area occurs, and a large amount of forsythia leaves are abandoned while the shortage of forsythia resources is caused, so that the resource waste is greatly caused.
The effective components of phillyrin, forsythiaside and the like contained in the forsythia suspense leaves are similar to those of forsythia suspense fruits, and the total amount of the phillyrin suspense leaves is more than 10 times of that of the fruits. Research results show that the forsythia suspense leaves have pharmacological activities of resisting aging, resisting oxidation, removing free radicals, resisting bacteria, viruses, infection and endotoxin, and the like, improve the growth performance and health condition of animals, and have low toxicity and no residue. At present, forsythia suspense leaves become a hotspot of research on novel feed additives and have great development value.
The traditional Chinese medicine micro-ecological animal feed additive is a biological preparation prepared by fermenting traditional Chinese medicines with beneficial microorganisms, has the effects of promoting the growth and development of immune organs of animals, improving the immune function and disease resistance of organisms, promoting the growth and development of the animals, shortening the breeding period, improving the feed conversion rate and the production performance, improving the product quality, enhancing the drug effect, reducing the breeding cost and the like, is safe and harmless, has no residue, and is not easy to generate drug resistance.
Therefore, the forsythia suspense leaf microecological preparation is developed by combining the modern microbial fermentation technology, the effect of deep excavation of forsythia suspense leaves is facilitated, the market demand of the production process of livestock and poultry products on novel feed additives is met, the novel feed additives which can ensure the healthy growth and development of livestock and poultry and cannot cause harm to human health are developed, and the novel feed additives have wide market application prospects.
Disclosure of Invention
The invention aims to provide a micro-ecological preparation based on forsythia suspense leaves and a preparation method of the micro-ecological preparation.
The micro-ecological preparation based on forsythia suspense leaves is prepared by taking dry forsythia suspense leaf powder as a raw material, inoculating bacillus subtilis and candida utilis to perform aerobic fermentation, and then inoculating lactobacillus bulgaricus of German type to perform anaerobic fermentation.
The forsythia suspense leaves have pharmacological activities of resisting aging, resisting oxidation, removing free radicals, protecting the liver, enhancing immunity, resisting stress and the like, can improve the growth performance and the health condition of animals, and have low toxicity and no residue. The forsythia suspense leaf microecological preparation is prepared by inoculating probiotics to the forsythia suspense leaf microecological preparation and fermenting, contains abundant beneficial microorganisms, can increase the number and variety of microorganisms in animal intestinal tracts, competitively reject pathogenic microorganisms, improve the intestinal tract function, improve the feed utilization rate and reduce the feed conversion ratio. In addition, the cell wall of forsythia suspense leaves can be destroyed by microorganisms during metabolism, which is helpful for the precipitation of active ingredients of forsythia suspense and enhances the drug effect of the forsythia suspense.
Furthermore, the invention also provides a specific preparation method of the forsythia suspense leaf-based microecological preparation, which is characterized in that the forsythia suspense leaf dry powder is used as a raw material, and bacillus subtilis (3-5) multiplied by 10 is inoculated according to each kilogram of forsythia suspense leaf dry powder10～(3～5)×10111-3 multiplied by 10 Candida utilis10～(4～7)×1011Aerobic fermentation is carried out for 48-72 h at 25-40 ℃, and then lactobacillus bulgaricus (5-8) multiplied by 10 is inoculated according to per kilogram of forsythia suspense leaf dry powder9～(5～8)×1010Sealing, and carrying out anaerobic fermentation at 35-40 ℃ for 24-72 h to prepare the forsythia suspense leaf micro-ecological preparation.
Furthermore, the aerobic fermentation is preferably performed for 24 hours at the temperature of 25-30 ℃ and then for 24-48 hours at the temperature of 35-40 ℃.
The forsythia suspense leaf dry powder is prepared by picking fresh forsythia suspense leaves, cleaning, naturally drying, crushing, sieving with a 40-mesh sieve, sterilizing and drying until the water content is below 6%.
Further, the fresh forsythia suspense leaves are preferably picked for 5-7 months.
The prepared forsythia suspense leaf microecological preparation is dried until the water content is 10-12%. The preferable drying temperature is 25-30 ℃.
The forsythia suspense leaf microecological preparation prepared by the method can be used as a livestock and poultry feed additive and directly added into livestock and poultry feed, and the addition amount is generally 1-5% of the weight of the feed.
After beneficial bacteria are added into the forsythia suspense leaves for fermentation, compared with the unfermented forsythia suspense leaves, the content of part of effective components such as forsythin and forsythiaside A is increased by 15-30%, and the utilization degree of forsythia suspense leaf resources is improved.
The forsythia suspense leaf microecological preparation prepared by the method is used as a livestock and poultry feed additive, is directly added into livestock and poultry feed to feed broiler chickens, can improve the utilization rate of the broiler chickens feed, effectively improve the production performance of the broiler chickens, reduce the diarrhea rate and the death rate of the broiler chickens, obviously improve the thymus index and the bursal disease index of the broiler chickens, increase the IgA and IgM contents in serum, enhance the immunologic function of the broiler chickens, has the efficacy superior to that of forsythia suspense leaves and basically equivalent to that of antibiotics, and can reduce the use of antibiotics.
The preparation method of the forsythia suspense leaf microecological preparation is simple and feasible, safe, environment-friendly and low in cost, and can meet the market demand of a novel feed additive in the production process of animal products.
Detailed Description
The following examples further describe embodiments of the present invention. The following examples are only for illustrating the technical solutions of the present invention more clearly, and do not limit the scope of the present invention. Various changes, modifications, substitutions and alterations to these embodiments will be apparent to those skilled in the art without departing from the principles and spirit of this invention.
Example 1.
Selecting a bacillus subtilis monoclonal colony preserved in a slant culture medium, inoculating the bacillus subtilis monoclonal colony in a liquid complete culture medium containing 0.5% of beef extract, 1% of peptone and 0.5% of sodium chloride, and culturing the bacillus subtilis monoclonal colony for 36-72 hours in a shaker at 25-35 ℃ at 100-160 rpm until the viable count is (3 multiplied by 10)8～3×109) One per ml.
Inoculating the candida utilis freeze-dried powder into YPD culture medium containing 2% of glucose, 2% of peptone and 1% of yeast powder, and culturing for 72-96 h in a shaker at 25-35 ℃ at 100-160 rpm until the viable count is (1 x 10)8～1×109) One per ml.
Preparing MRS culture medium containing 1% of peptone, 0.8% of beef extract, 0.4% of yeast powder, 2.2% of glucose, 0.04% of magnesium sulfate, 0.8% of sodium acetate, 0.2% of triammonium citrate, 1% of dipotassium phosphate, 0.025% of manganese sulfate and 800.1% of tween, inoculating freeze-dried powder of Lactobacillus bulgaricus, culturing for 48h at the temperature of 35-40 ℃ and the rpm of 100-160 in a shaking table until the viable count is (6 multiplied by 10) to reach (6 multiplied by 10)7～6×108) One per ml.
And in 6-7 months, picking fresh forsythia suspense leaves, cleaning, naturally airing, crushing, sieving by using a 40-mesh sieve, performing damp-heat sterilization at 121 ℃, and drying in an oven at 45-55 ℃ until the water content is below 6% to prepare the forsythia suspense leaf dry powder.
Under aseptic operation, inoculating bacillus subtilis (3-5) x 10 to each kilogram of forsythia suspense leaf dry powder101-3 multiplied by 10 Candida utilis10Fully and uniformly stirring, putting into a fermentation tank, controlling the temperature to be 25-30 ℃ for fermentation for 24 hours, controlling the temperature to be 35-40 ℃, and continuing to ferment for 48 hours.
After fermentation is finished, inoculating lactobacillus bulgaricus (5-8) multiplied by 10 per kilogram of forsythia suspense leaf dry powder9Fully and uniformly stirring, filling into a fermentation tank, sealing, controlling the temperature at 35-40 ℃, and carrying out anaerobic fermentation for 24-48 h.
And after fermentation, drying at 25-30 ℃ until the water content is 10-12%, and preparing the forsythia suspense leaf microecological preparation.
Example 2.
The dry powder of forsythia suspense leaves prepared in example 1 is used for inoculating bacillus subtilis (3-5) multiplied by 10 cultured in example 1 per kilogram of dry powder of forsythia suspense leaves under aseptic operation11Candida utilis (4-7) x 1011Fully and uniformly stirring, putting into a fermentation tank, controlling the temperature to be 32-37 ℃ for fermentation for 24h, and then controlling the temperature to be 35-40 ℃ for continuous fermentation for 24 h.
After the fermentation is finished, the cultured Lactobacillus bulgaricus (5-8). times.10 of the cultured Lactobacillus bulgaricus in example 1 is inoculated to each kilogram of the forsythia suspense leaf dry powder10Fully and uniformly stirring, filling into a fermentation tank, sealing, controlling the temperature at 35-40 ℃, and performing anaerobic fermentation for 72 hours.
And after fermentation, drying at 25-30 ℃ until the water content is 10-12%, and preparing the forsythia suspense leaf microecological preparation.
Application example: the influence of the forsythia suspense leaf micro-ecological feed additive on the growth performance and the immune function of the broiler chicken.
800 healthy broiler chickens of 1 day old are selected and randomly divided into a blank control group, an antibiotic group, a forsythia suspensa leaf group and a forsythia suspensa leaf microecological preparation group, wherein 200 broiler chickens are selected.
The blank control group was fed basal diet only. On the basis of feeding basic daily ration for other groups, 100mg/kg of aureomycin is added in an antibiotic group, 3% of dry powder of forsythia leaves is added in a forsythia leaves group, and 3% of microecological preparation of forsythia leaves is added in a forsythia leaves microecological preparation group. The feed is fed once every morning and afternoon.
The basic ration comprises the following components: 60.3% of corn, 4% of bran, 4% of sunflower seed cake, 6% of cottonseed cake, 18% of soybean cake, 5% of fish meal, 1.5% of bone meal, 1% of shell powder and 0.2% of salt.
The experimental broiler chickens were raised in cages of 8 chickens per cage, and were fed and drunk freely, and immunized according to the conventional broiler chicken procedure.
Weighing before experiment, and recording feed intake of each group.
The experiment lasts for 42 days, the phenomenon that excrement of the broiler chicken is applied to the anus due to diarrhea is recorded every day, the diarrhea rate is counted, the death number of the broiler chicken is observed, and the death rate is calculated.
On the 42 th day of the experiment, 100 broilers were randomly selected from each group, and the daily gain and the feed-weight ratio were calculated. Bleeding and killing jugular vein, collecting blood sample, preparing serum, and detecting IgA and IgM contents in the serum by an immunoturbidimetry method; quickly picking up thymus and bursa of Fabricius, wiping off surface water, weighing and calculating immune organ index.
The results in table 1 show that the forsythia suspense leaf microecological preparation can obviously reduce death of broiler chickens, inhibit diarrhea, increase daily gain of broiler chickens and reduce feed conversion ratio compared with a blank control group. Compared with the forsythia suspense leaf group, the forsythia suspense leaf microecological preparation can further improve the growth performance of broiler chickens, and compared with the antibiotic group, the forsythia suspense leaf microecological preparation has the effect of improving the growth performance of broiler chickens similar to that of antibiotics.
As can be seen from Table 2, the forsythia suspense leaf microecological preparation can obviously increase the thymus index, the bursa of Fabricius index and the IgA and IgM contents in serum of the broiler chicken compared with the blank control group. Compared with the forsythia suspense leaf group, the forsythia suspense leaf microecological preparation can further enhance the immune function of the broiler chicken, and compared with the antibiotic group, the forsythia suspense leaf microecological preparation has the effect of enhancing the immune function similar to that of the antibiotic.
Claims (9)
1. A micro-ecological preparation based on weeping forsythia leaves is a micro-ecological preparation obtained by taking weeping forsythia leaf dry powder as a raw material, inoculating bacillus subtilis and candida utilis to perform aerobic fermentation, and then inoculating lactobacillus bulgaricus of German type to perform anaerobic fermentation.
2. The method for preparing a micro-ecological preparation based on forsythia suspense leaves as claimed in claim 1, wherein the method comprises inoculating bacillus subtilis (3-5) x 10 to each kilogram of forsythia suspense leaf dry powder as a raw material10～(3～5)×10111-3 multiplied by 10 Candida utilis10～(4～7)×1011Aerobic fermentation is carried out for 48-72 h at 25-40 ℃, and then lactobacillus bulgaricus (5-8) multiplied by 10 is inoculated according to per kilogram of forsythia suspense leaf dry powder9～(5～8)×1010Sealing, and carrying out anaerobic fermentation at 35-40 ℃ for 24-72 h to prepare the forsythia suspense leaf micro-ecological preparation.
3. The method according to claim 2, wherein the aerobic fermentation is performed at a temperature controlled at 25-30 ℃ for 24 hours, and then at a temperature controlled at 35-40 ℃ for 24-48 hours.
4. The preparation method according to claim 2, wherein the dry powder of forsythia suspense leaves is prepared by picking fresh forsythia suspense leaves, cleaning, naturally drying, crushing and sieving with a 40-mesh sieve, sterilizing and drying until the water content is below 6%.
5. The method according to claim 4, wherein the fresh forsythia suspense leaves are forsythia suspense leaves picked in 5 to 7 months.
6. The preparation method of claim 2, wherein the prepared forsythia suspense leaf microecological preparation is dried to a water content of 10-12%.
7. The method according to claim 6, wherein the drying temperature is 25 to 30 ℃.
8. Use of the forsythia suspense leaf-based microecological preparation according to claim 1 as a feed additive for livestock and poultry.
9. The use of claim 8, wherein the additive is added into the livestock and poultry feed in an amount of 1-5% by weight of the feed.
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| CN113546111A (en) * | 2021-07-30 | 2021-10-26 | 山西中医药大学 | Microbial fermentation method for improving content of effective components in forsythia suspense leaves |
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