CN113519718A - Broiler chicken probiotics feed additive and preparation method and application thereof - Google Patents
Broiler chicken probiotics feed additive and preparation method and application thereof Download PDFInfo
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- CN113519718A CN113519718A CN202110784338.0A CN202110784338A CN113519718A CN 113519718 A CN113519718 A CN 113519718A CN 202110784338 A CN202110784338 A CN 202110784338A CN 113519718 A CN113519718 A CN 113519718A
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- enterococcus faecium
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- 241000287828 Gallus gallus Species 0.000 title claims abstract description 51
- 239000003674 animal food additive Substances 0.000 title claims abstract description 48
- 239000006041 probiotic Substances 0.000 title claims abstract description 43
- 235000018291 probiotics Nutrition 0.000 title claims abstract description 43
- 238000002360 preparation method Methods 0.000 title claims abstract description 21
- 239000000843 powder Substances 0.000 claims abstract description 81
- 241000194031 Enterococcus faecium Species 0.000 claims abstract description 70
- 238000000855 fermentation Methods 0.000 claims abstract description 58
- 230000004151 fermentation Effects 0.000 claims abstract description 58
- 239000007787 solid Substances 0.000 claims abstract description 36
- 241000209094 Oryza Species 0.000 claims abstract description 33
- 235000007164 Oryza sativa Nutrition 0.000 claims abstract description 33
- 235000009566 rice Nutrition 0.000 claims abstract description 33
- 239000002068 microbial inoculum Substances 0.000 claims abstract description 30
- 241000512259 Ascophyllum nodosum Species 0.000 claims abstract description 23
- 241000194108 Bacillus licheniformis Species 0.000 claims abstract description 22
- 230000000529 probiotic effect Effects 0.000 claims abstract description 22
- 241000194103 Bacillus pumilus Species 0.000 claims abstract description 21
- 244000063299 Bacillus subtilis Species 0.000 claims abstract description 21
- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 21
- 229910021536 Zeolite Inorganic materials 0.000 claims abstract description 19
- HNPSIPDUKPIQMN-UHFFFAOYSA-N dioxosilane;oxo(oxoalumanyloxy)alumane Chemical compound O=[Si]=O.O=[Al]O[Al]=O HNPSIPDUKPIQMN-UHFFFAOYSA-N 0.000 claims abstract description 19
- FTSSQIKWUOOEGC-RULYVFMPSA-N fructooligosaccharide Chemical compound OC[C@H]1O[C@@](CO)(OC[C@@]2(OC[C@@]3(OC[C@@]4(OC[C@@]5(OC[C@@]6(OC[C@@]7(OC[C@@]8(OC[C@@]9(OC[C@@]%10(OC[C@@]%11(O[C@H]%12O[C@H](CO)[C@@H](O)[C@H](O)[C@H]%12O)O[C@H](CO)[C@@H](O)[C@@H]%11O)O[C@H](CO)[C@@H](O)[C@@H]%10O)O[C@H](CO)[C@@H](O)[C@@H]9O)O[C@H](CO)[C@@H](O)[C@@H]8O)O[C@H](CO)[C@@H](O)[C@@H]7O)O[C@H](CO)[C@@H](O)[C@@H]6O)O[C@H](CO)[C@@H](O)[C@@H]5O)O[C@H](CO)[C@@H](O)[C@@H]4O)O[C@H](CO)[C@@H](O)[C@@H]3O)O[C@H](CO)[C@@H](O)[C@@H]2O)[C@@H](O)[C@@H]1O FTSSQIKWUOOEGC-RULYVFMPSA-N 0.000 claims abstract description 19
- 229940107187 fructooligosaccharide Drugs 0.000 claims abstract description 19
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- 238000002156 mixing Methods 0.000 claims abstract description 11
- 238000003756 stirring Methods 0.000 claims abstract description 7
- 239000002994 raw material Substances 0.000 claims abstract description 5
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims description 64
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 64
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 48
- 229910000019 calcium carbonate Inorganic materials 0.000 claims description 32
- 229940041514 candida albicans extract Drugs 0.000 claims description 32
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 32
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 32
- 229940099596 manganese sulfate Drugs 0.000 claims description 32
- 239000011702 manganese sulphate Substances 0.000 claims description 32
- 235000007079 manganese sulphate Nutrition 0.000 claims description 32
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 32
- 239000012138 yeast extract Substances 0.000 claims description 32
- 239000007788 liquid Substances 0.000 claims description 30
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 24
- 239000001888 Peptone Substances 0.000 claims description 24
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 24
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- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 16
- 229930006000 Sucrose Natural products 0.000 claims description 16
- 238000012258 culturing Methods 0.000 claims description 16
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 16
- 238000001035 drying Methods 0.000 claims description 16
- 239000000203 mixture Substances 0.000 claims description 16
- 238000011218 seed culture Methods 0.000 claims description 16
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- 239000005720 sucrose Substances 0.000 claims description 14
- 235000013406 prebiotics Nutrition 0.000 claims description 13
- 238000009630 liquid culture Methods 0.000 claims description 12
- 238000010563 solid-state fermentation Methods 0.000 claims description 12
- 230000003716 rejuvenation Effects 0.000 claims description 10
- 238000011081 inoculation Methods 0.000 claims description 9
- 229920001817 Agar Polymers 0.000 claims description 8
- 239000008272 agar Substances 0.000 claims description 8
- YXVFQADLFFNVDS-UHFFFAOYSA-N diammonium citrate Chemical compound [NH4+].[NH4+].[O-]C(=O)CC(O)(C(=O)O)CC([O-])=O YXVFQADLFFNVDS-UHFFFAOYSA-N 0.000 claims description 8
- 235000001727 glucose Nutrition 0.000 claims description 7
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- 238000010298 pulverizing process Methods 0.000 claims description 4
- 235000015097 nutrients Nutrition 0.000 claims description 3
- 235000015099 wheat brans Nutrition 0.000 claims description 3
- 244000046052 Phaseolus vulgaris Species 0.000 claims description 2
- 235000010627 Phaseolus vulgaris Nutrition 0.000 claims description 2
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims description 2
- 229920000053 polysorbate 80 Polymers 0.000 claims description 2
- 235000010216 calcium carbonate Nutrition 0.000 claims 2
- 210000001035 gastrointestinal tract Anatomy 0.000 abstract description 8
- 241001465754 Metazoa Species 0.000 abstract description 7
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- 230000000052 comparative effect Effects 0.000 description 12
- 235000013330 chicken meat Nutrition 0.000 description 10
- 230000000694 effects Effects 0.000 description 6
- 239000000126 substance Substances 0.000 description 5
- 238000004321 preservation Methods 0.000 description 4
- 238000007873 sieving Methods 0.000 description 4
- 239000000654 additive Substances 0.000 description 3
- 230000000996 additive effect Effects 0.000 description 3
- 239000003242 anti bacterial agent Substances 0.000 description 3
- 229940088710 antibiotic agent Drugs 0.000 description 3
- 238000009395 breeding Methods 0.000 description 3
- 230000001488 breeding effect Effects 0.000 description 3
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- 239000000463 material Substances 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 235000016709 nutrition Nutrition 0.000 description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- 208000035240 Disease Resistance Diseases 0.000 description 2
- 206010059866 Drug resistance Diseases 0.000 description 2
- 235000010443 alginic acid Nutrition 0.000 description 2
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- 238000002474 experimental method Methods 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- 244000144972 livestock Species 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
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- 230000004584 weight gain Effects 0.000 description 2
- 235000019786 weight gain Nutrition 0.000 description 2
- FHVDTGUDJYJELY-UHFFFAOYSA-N 6-{[2-carboxy-4,5-dihydroxy-6-(phosphanyloxy)oxan-3-yl]oxy}-4,5-dihydroxy-3-phosphanyloxane-2-carboxylic acid Chemical compound O1C(C(O)=O)C(P)C(O)C(O)C1OC1C(C(O)=O)OC(OP)C(O)C1O FHVDTGUDJYJELY-UHFFFAOYSA-N 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- 241000282414 Homo sapiens Species 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 241000186840 Lactobacillus fermentum Species 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 229940072056 alginate Drugs 0.000 description 1
- 229910001579 aluminosilicate mineral Inorganic materials 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 230000002860 competitive effect Effects 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- 230000001934 delay Effects 0.000 description 1
- 230000001079 digestive effect Effects 0.000 description 1
- FPAFDBFIGPHWGO-UHFFFAOYSA-N dioxosilane;oxomagnesium;hydrate Chemical compound O.[Mg]=O.[Mg]=O.[Mg]=O.O=[Si]=O.O=[Si]=O.O=[Si]=O.O=[Si]=O FPAFDBFIGPHWGO-UHFFFAOYSA-N 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
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- 239000003814 drug Substances 0.000 description 1
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- 238000013401 experimental design Methods 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- PNDPGZBMCMUPRI-UHFFFAOYSA-N iodine Chemical compound II PNDPGZBMCMUPRI-UHFFFAOYSA-N 0.000 description 1
- 238000005342 ion exchange Methods 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 229940012969 lactobacillus fermentum Drugs 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 235000013622 meat product Nutrition 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
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- 231100000252 nontoxic Toxicity 0.000 description 1
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- 239000011574 phosphorus Substances 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
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- 244000144977 poultry Species 0.000 description 1
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- 230000004083 survival effect Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
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Classifications
-
- 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
-
- 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
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
-
- 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
- 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
- A23K10/37—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms from waste material
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/163—Sugars; Polysaccharides
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/20—Inorganic substances, e.g. oligoelements
- A23K20/28—Silicates, e.g. perlites, zeolites or bentonites
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/36—Adaptation or attenuation of cells
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
Abstract
The invention discloses a broiler chicken probiotics feed additive and a preparation method and application thereof, and belongs to the technical field of feed additives. The raw materials of the probiotic feed additive comprise: bacillus subtilis, bacillus licheniformis, bacillus pumilus, enterococcus faecium solid fermentation microbial inoculum, fructooligosaccharide, kelp powder, zeolite powder and rice bran. The preparation method comprises the steps of preparing the enterococcus faecium into a solid fermentation microbial inoculum, uniformly mixing the solid fermentation microbial inoculum with the bacillus subtilis, the bacillus licheniformis, the bacillus pumilus, the fructooligosaccharide and the zeolite powder, then adding the kelp powder and the crushed rice bran, and uniformly stirring to obtain the probiotic feed additive. The strain disclosed by the invention is reasonable in compounding and strong in stress resistance, and after the strain is added into the feed, the strain is not easy to inactivate after entering animal intestinal tracts, so that the absorption conversion rate of the probiotic feed can be effectively improved, and the weight increasing effect of the broiler chicken is remarkable.
Description
Technical Field
The invention relates to the technical field of feed additives, in particular to a broiler chicken probiotics feed additive and a preparation method and application thereof.
Background
In recent years, with the increasing living standard of people, the demand of meat products is increasing, but in the broiler breeding, in order to shorten the breeding period, reduce the death rate and reduce the cost, most chicken farmers often choose to add antibiotics and other medicines into the feed. In the past, the chicken has drug resistance and drug resistance in the breeding process, endogenous infection occurs, and meanwhile, the problems that the immunologic function of the chicken is reduced, the disease resistance is reduced, antibiotics are left in the chicken and the like exist.
The probiotics is a bioactive preparation prepared from living microorganisms, and can inhibit the growth of harmful bacteria through the competitive exclusion action of animal digestive tract organisms, form a dominant flora or prevent diseases by enhancing the nonspecific immunity function, thereby promoting the growth of animals and improving the feed conversion rate. The probiotic feed additive is non-toxic to livestock products, has no side effect on human beings, and is a novel green feed additive developed in the last ten years.
Although probiotics have the advantages of no toxicity, no side effect and the like compared with antibiotics, the probiotics feed additive still has the problem of poor strain stress resistance, so that strains are easy to inactivate after the feed enters animal intestinal tracts, the strains are difficult to better exert effects, and meanwhile, the absorption cyclic cyclization rate of the probiotics feed is low and the weight gain of broiler chickens is difficult to meet the requirements due to factors such as unreasonable strain compounding.
Disclosure of Invention
The invention aims to provide a broiler chicken probiotics feed additive and a preparation method and application thereof, and aims to solve the problems in the prior art.
In order to achieve the purpose, the invention provides the following scheme:
the invention provides a broiler chicken probiotics feed additive which comprises the following raw materials in parts by weight:
0.8-1.5 parts of bacillus subtilis, 0.5-1.8 parts of bacillus licheniformis, 0.6-1.2 parts of bacillus pumilus, 1.3-1.8 parts of enterococcus faecium solid fermentation microbial inoculum, 5-8 parts of fructooligosaccharide, 20-30 parts of kelp powder, 25-35 parts of zeolite powder and 50-60 parts of rice bran;
the preparation method of the enterococcus faecium solid fermentation microbial inoculum comprises the following steps:
a. screening and domesticating the strains: under the aseptic operation condition, putting enterococcus faecium into an improved MRS culture medium containing glucose, peptone, beef extract, yeast extract powder, sodium chloride, agar powder, calcium carbonate, diammonium citrate, Tween 80, magnesium sulfate and manganese sulfate, streaking, culturing, selecting typical enterococcus faecium single colonies, streaking into the MRS culture medium respectively, and continuously purifying, subculturing and rejuvenating;
b. primary seed liquid culture: under the aseptic operation condition, inoculating the screened and domesticated enterococcus faecium single colony into a primary seed culture solution containing sucrose, glucose, peptone, calcium carbonate, yeast extract powder, sodium chloride, beef extract, magnesium sulfate, manganese sulfate and dipotassium hydrogen phosphate, carrying out shake culture, obtaining a primary seed solution after finishing the culture, and storing the primary seed solution in a refrigerator at 4 ℃ for later use;
c. secondary seed liquid culture: inoculating the prepared primary seed liquid into a secondary seed culture liquid containing cane sugar, glucose, peptone, calcium carbonate, yeast extract powder, sodium chloride, beef extract, magnesium sulfate, manganese sulfate and dipotassium hydrogen phosphate in a sterile environment, and culturing in a seed tank to obtain a secondary seed liquid;
d. solid state fermentation: uniformly inoculating the prepared secondary seed liquid into a solid fermentation culture medium containing bean pulp, rice bran, wheat bran, calcium carbonate, yeast extract powder, magnesium sulfate and manganese sulfate for fermentation culture in an aseptic environment;
e. drying and crushing: and drying the fermentation product after the fermentation is finished, and crushing to obtain the enterococcus faecium solid fermentation microbial inoculum.
Further, the raw materials comprise the following components in parts by weight:
1.2 parts of bacillus subtilis, 1.3 parts of bacillus licheniformis, 0.8 part of bacillus pumilus, 1.5 parts of enterococcus faecium solid fermentation microbial inoculum, 6 parts of fructooligosaccharide, 25 parts of kelp powder, 30 parts of zeolite powder and 55 parts of rice bran.
Further, in the step a, the modified MRS culture medium contains 2.5-2.6% of glucose, 1.3-1.5% of peptone, 1.3% of beef extract, 1.2-1.5% of yeast extract powder, 0.8% of sodium chloride, 3.1-3.3% of agar powder, 2.5-2.7% of calcium carbonate, 1% of diammonium citrate, 800.1-0.2% of Tween, 0.1% of magnesium sulfate, 0.06% of manganese sulfate and the balance of purified water;
in the step b, the primary seed culture solution contains 2.3-2.4% of sucrose, 2-2.1% of glucose, 1.6-1.8% of peptone, 5.2-5.4% of calcium carbonate, 1.5% of yeast extract powder, 0.8% of sodium chloride, 1.8-2.0% of beef extract, 0.8% of magnesium sulfate, 0.8% of manganese sulfate, 3.3-3.4% of dipotassium hydrogen phosphate and the balance of purified water; in the step b, the temperature of the shaking culture is 35-40 ℃, and the time is 72-74 h.
Further, in the step c, the secondary seed nutrient solution contains 2.8-3.0% of sucrose, 1.3-1.5% of glucose, 1.8-2.0% of peptone, 4.3-4.5% of calcium carbonate, 1% of yeast extract powder, 0.9% of sodium chloride, 1.7% of beef extract, 0.8% of magnesium sulfate, 0.8-1.2% of manganese sulfate, 3.3% of dipotassium hydrogen phosphate and the balance of purified water;
in the step c, the inoculation amount of the primary seed liquid is 8-10% of the volume of the secondary seed culture liquid, the temperature of the seed tank culture is 35-40 ℃, and the time is 72-74 h.
Further, in the step d, the solid fermentation medium contains 40-43% of soybean meal, 12-13% of rice bran, 23-25% of bran, 11-15% of calcium carbonate, 2.7-2.8% of yeast extract powder, 1.2% of magnesium sulfate, 1.2% of manganese sulfate and the balance of purified water;
in the step d, the inoculation amount of the secondary seed liquid is 15-20% of the volume of the solid fermentation culture medium, the fermentation culture temperature is 35-40 ℃, and the fermentation culture time is 68-72 hours.
Further, in the step e, the drying temperature is 30-50 ℃, the drying time is 3-6h, and the crushed meshes are 80-100 meshes.
The invention also provides a preparation method of the broiler chicken probiotics feed additive, which comprises the following steps:
(1) pulverizing testa oryzae;
(2) uniformly mixing a solid fermentation microbial inoculum of bacillus subtilis, bacillus licheniformis, bacillus pumilus and enterococcus faecium with fructooligosaccharide and zeolite powder to obtain a mixture;
(3) and (3) adding kelp powder and crushed rice bran into the mixture obtained in the step (2), and uniformly stirring to obtain the prebiotics feed additive.
Further, in the step (1), the rice bran is crushed and then sieved by a 60-80 mesh sieve.
The invention also provides an application of the broiler chicken probiotics feed additive in feed.
Furthermore, the addition amount of the broiler chicken probiotics feed additive in the feed is 0.9-1.2% of the total weight of the feed.
The invention discloses the following technical effects:
1. according to the invention, the enterococcus faecium, the bacillus subtilis, the bacillus licheniformis and the bacillus pumilus are compounded and then added into the probiotic additive, the strain is reasonably compounded, the absorption conversion rate of the probiotic feed can be improved, and the broiler weight increasing effect is obvious.
2. The enterococcus faecium solid fermentation microbial inoculum has good effects on the aspects of increasing weight gain of young livestock and poultry, improving animal immunity, adjusting intestinal microecological balance, improving nutrition absorption and the like, but has poor stress resistance, and can not effectively maintain activity in feed production and processing, thereby influencing the effect. The enterococcus faecium is domesticated and screened, the stress resistance of the enterococcus faecium is obviously improved, meanwhile, bacillus subtilis, bacillus licheniformis and bacillus pumilus belong to the same genus of bacillus, the stress resistance of the bacillus subtilis, the bacillus licheniformis and the bacillus pumilus is strong, high activity can be kept in animal intestinal tracts, the strains are guaranteed to have strong stress resistance on the whole, and the survival rate of the strains after entering the animal intestinal tracts is improved.
3. The kelp powder is rich in nutritional ingredients such as iodine, algal polysaccharides, mannitol and the like and various bioactive substances, and can improve the immunity of the broiler chicken and obviously improve the meat quality; the kelp powder is also rich in algin substances, and can form a layer of alginate film on the surface of the feed, thereby increasing the adhesion between the additive and the feed and improving the utilization rate of the additive.
4. According to the invention, the fructo-oligosaccharide is added into the probiotic feed additive, the fructo-oligosaccharide is preferentially utilized by the probiotics after entering the digestive tract of the broiler chicken, and in the proliferation process of the probiotics, a large amount of substances beneficial to the proliferation and growth of the probiotics can be generated, so that the mass propagation of the probiotics is promoted, the competitiveness of the probiotics is greatly enhanced, and the beneficial flora is rapidly activated.
5. The zeolite powder is aluminosilicate mineral, contains various major and trace elements such as calcium, phosphorus, potassium, sodium and the like, can improve the nutritional value of the feed, improve the disease resistance of organisms and promote the growth of broilers; meanwhile, the zeolite powder also has good adsorbability and ion exchange property, absorbs ammonia and fixes nitrogen, delays the time of nutrient substances passing through the digestive tract, adsorbs harmful substances in the intestinal tract, and improves the digestive function.
Detailed Description
Reference will now be made in detail to various exemplary embodiments of the invention, the detailed description should not be construed as limiting the invention but as a more detailed description of certain aspects, features and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. Further, for numerical ranges in this disclosure, it is understood that each intervening value, between the upper and lower limit of that range, is also specifically disclosed. Every smaller range between any stated value or intervening value in a stated range and any other stated or intervening value in a stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference herein for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the present disclosure without departing from the scope or spirit of the disclosure. Other embodiments will be apparent to those skilled in the art from consideration of the specification. The specification and examples are exemplary only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are open-ended terms that mean including, but not limited to.
The "parts" in the present invention are all parts by mass unless otherwise specified.
The preservation number of the bacillus subtilis used in the invention is CGMCC No.3665, and the bacillus subtilis is purchased from the common microorganism center of China Committee for culture Collection of microorganisms;
the preservation number of the used bacillus licheniformis is CGMCC No.7388, and the bacillus licheniformis is purchased from the China general microbiological culture Collection center;
the preservation number of the used bacillus pumilus is CCTCC No. M2014008, and the bacillus pumilus is purchased from China center for type culture collection;
the preservation number of the enterococcus faecium is CGMCC No.7274, and the enterococcus faecium is purchased from the common microorganism center of China Committee for culture Collection of microorganisms.
Example 1
1) Preparing an enterococcus faecium solid fermentation microbial inoculum:
a. screening and domesticating the strains: under the aseptic operation condition, putting enterococcus faecium into an improved MRS culture medium containing 2.5% of glucose, 1.4% of peptone, 1.3% of beef extract, 1.3% of yeast extract powder, 0.8% of sodium chloride, 3.2% of agar powder, 2.6% of calcium carbonate, 1% of diammonium citrate, 800.1% of tween, 0.1% of magnesium sulfate, 0.06% of manganese sulfate and the balance of purified water, streaking, culturing at 37 ℃ for 24 hours, selecting 3-5 typical enterococcus faecium single colonies, streaking into the MRS culture medium respectively, continuously purifying, carrying out passage rejuvenation for 3 times, finally selecting the rejuvenated enterococcus faecium for physiological and biochemical identification, and selecting the optimal enterococcus faecium single colony as a seed strain to serve as a primary seed solution strain after the enterococcus faecium is identified as enterococcus faecium;
b. primary seed liquid culture: under the aseptic operation condition, selecting single colonies of screened and domesticated enterococcus faecium, inoculating the single colonies into a primary seed culture solution which comprises 2.3% of sucrose, 2% of glucose, 1.6% of peptone, 5.2% of calcium carbonate, 1.5% of yeast extract powder, 0.8% of sodium chloride, 2.0% of beef extract, 0.8% of magnesium sulfate, 0.8% of manganese sulfate, 3.4% of dipotassium hydrogen phosphate and the balance of purified water, culturing for 72 hours at the temperature of 40 ℃ in a shaking table at the rotating speed of 250r/min, obtaining a primary seed solution after the culture is finished, and storing the primary seed solution in a refrigerator at the temperature of 4 ℃ for later use;
c. secondary seed liquid culture: inoculating the prepared primary seed liquid into a secondary seed culture liquid containing 2.9% of sucrose, 1.4% of glucose, 1.90% of peptone, 4.4% of calcium carbonate, 1% of yeast extract powder, 0.9% of sodium chloride, 1.7% of beef extract, 0.8% of magnesium sulfate, 1.0% of manganese sulfate, 3.3% of dipotassium hydrogen phosphate and the balance of purified water under an aseptic environment by the inoculation amount of 10% v/v, and culturing for 73 hours at the temperature of 38 ℃ in a seed tank at the rotating speed of 320r/min to obtain a secondary seed liquid;
d. solid state fermentation: under an aseptic environment, the prepared secondary seed liquid is uniformly inoculated into a solid fermentation culture medium containing 43% of soybean meal, 13% of rice bran, 23% of wheat bran, 11% of calcium carbonate, 2.7% of yeast extract powder, 1.2% of magnesium sulfate, 1.2% of manganese sulfate and the balance of purified water in an inoculation amount of 20% v/v, and fermentation culture is carried out for 68 hours at 35 ℃.
e. Drying and crushing: and after the fermentation is finished, drying the fermentation product at 50 ℃ for 6 hours, and then crushing to 100 meshes to obtain the enterococcus faecium solid state fermentation microbial inoculum.
2) Preparation of probiotic feed additive
1.2 parts of bacillus subtilis, 1.3 parts of bacillus licheniformis, 0.8 part of bacillus pumilus, 1.5 parts of enterococcus faecium solid fermentation microbial inoculum, 6 parts of fructooligosaccharide, 25 parts of kelp powder, 30 parts of zeolite powder and 55 parts of rice bran.
(1) Crushing rice bran and sieving with a 70-mesh sieve;
(2) mixing the solid fermentation microbial inoculum of bacillus subtilis, bacillus licheniformis, bacillus pumilus and enterococcus faecium with fructooligosaccharide and zeolite powder at the rotation speed of 250r/min for 3min, and mixing uniformly to obtain a mixture;
(3) keeping the rotating speed unchanged, adding the kelp powder and the crushed rice bran into the mixture obtained in the step (2), and stirring for 5min to uniformly mix the kelp powder and the crushed rice bran to obtain the broiler probiotic feed additive.
Example 2
1) Preparing an enterococcus faecium solid fermentation microbial inoculum:
a. screening and domesticating the strains: under the aseptic operation condition, putting enterococcus faecium into an improved MRS culture medium containing 2.6% of glucose, 1.5% of peptone, 1.3% of beef extract, 1.5% of yeast extract powder, 0.8% of sodium chloride, 3.1% of agar powder, 2.7% of calcium carbonate, 1% of diammonium citrate, 800.2% of tween, 0.1% of magnesium sulfate, 0.06% of manganese sulfate and the balance of purified water, streaking, culturing at 37 ℃ for 73h, selecting 3-5 typical enterococcus faecium single colonies, streaking into the MRS culture medium respectively, continuously purifying, carrying out subculture and rejuvenation for 3 times, finally selecting the rejuvenated enterococcus faecium for physiological and biochemical identification, and selecting the optimal enterococcus faecium single colony as a seed strain to serve as a first-level seed solution strain after the enterococcus faecium is identified as enterococcus faecium;
b. primary seed liquid culture: under the aseptic operation condition, selecting single colonies of screened and domesticated enterococcus faecium, inoculating the single colonies into a primary seed culture solution which comprises 2.3% of sucrose, 2% of glucose, 1.7% of peptone, 5.3% of calcium carbonate, 1.5% of yeast extract powder, 0.8% of sodium chloride, 1.9% of beef extract, 0.8% of magnesium sulfate, 0.8% of manganese sulfate, 3.3% of dipotassium hydrogen phosphate and the balance of purified water, culturing for 73 hours at 35 ℃ in a shaking table with the rotating speed of 180r/min, obtaining a primary seed solution after the culture is finished, and storing the primary seed solution in a refrigerator at 4 ℃ for later use;
c. secondary seed liquid culture: inoculating the prepared primary seed liquid into a secondary seed culture liquid containing 2.8% of sucrose, 1.3% of glucose, 1.8% of peptone, 4.3% of calcium carbonate, 1% of yeast extract powder, 0.9% of sodium chloride, 1.7% of beef extract, 0.8% of magnesium sulfate, 0.8% of manganese sulfate, 3.3% of dipotassium hydrogen phosphate and the balance of purified water in an inoculation amount of 9% v/v under a sterile environment, and culturing at 35 ℃ for 72 hours in a seed tank at the rotating speed of 350r/min to obtain a secondary seed liquid;
d. solid state fermentation: under the aseptic environment, the prepared secondary seed liquid is evenly inoculated into a solid fermentation culture medium containing 42% of soybean meal, 12% of rice bran, 24% of bran, 13% of calcium carbonate, 2.8% of yeast extract powder, 1.2% of magnesium sulfate, 1.2% of manganese sulfate and the balance of purified water, and fermentation culture is carried out for 72h at 38 ℃.
e. Drying and crushing: and after the fermentation is finished, drying the fermentation product at 30 ℃ for 3h, and then crushing to 80 meshes to obtain the enterococcus faecium solid state fermentation microbial inoculum.
2) Preparation of probiotic feed additive
0.8 part of bacillus subtilis, 0.5 part of bacillus licheniformis, 1.2 parts of bacillus pumilus, 1.8 parts of enterococcus faecium solid fermentation microbial inoculum, 7 parts of fructooligosaccharide, 20 parts of kelp powder, 30 parts of zeolite powder and 50 parts of rice bran.
(1) Pulverizing testa oryzae, and sieving with 60 mesh sieve;
(2) mixing the solid fermentation microbial inoculum of bacillus subtilis, bacillus licheniformis, bacillus pumilus and enterococcus faecium with fructo-oligosaccharide and zeolite powder at the rotating speed of 290r/min for 4min, and mixing uniformly to obtain a mixture;
(3) keeping the rotating speed unchanged, adding the kelp powder and the crushed rice bran into the mixture obtained in the step (2), and stirring for 6min to uniformly mix the kelp powder and the crushed rice bran to obtain the broiler probiotic feed additive.
Example 3
1) Preparing an enterococcus faecium solid fermentation microbial inoculum:
a. screening and domesticating the strains: under the aseptic operation condition, putting enterococcus faecium into an improved MRS culture medium containing 2.5% of glucose, 1.3% of peptone, 1.3% of beef extract, 1.2% of yeast extract powder, 0.8% of sodium chloride, 3.1% of agar powder, 2.5% of calcium carbonate, 1% of diammonium citrate, 800.1% of tween, 0.1% of magnesium sulfate, 0.06% of manganese sulfate and the balance of purified water, streaking, culturing at 37 ℃ for 24 hours, selecting 3-5 typical enterococcus faecium single colonies, streaking into the MRS culture medium respectively, continuously purifying, carrying out passage rejuvenation for 3 times, finally selecting the rejuvenated enterococcus faecium for physiological and biochemical identification, and selecting the optimal enterococcus faecium single colony as a seed strain to serve as a primary seed solution strain after the enterococcus faecium is identified as enterococcus faecium;
b. primary seed liquid culture: under the aseptic operation condition, selecting single colonies of screened and domesticated enterococcus faecium, inoculating the single colonies into a primary seed culture solution which comprises 2.3% of sucrose, 2.1% of glucose, 1.7% of peptone, 5.2% of calcium carbonate, 1.5% of yeast extract powder, 0.8% of sodium chloride, 1.9% of beef extract, 0.8% of magnesium sulfate, 0.8% of manganese sulfate, 3.4% of dipotassium hydrogen phosphate and the balance of purified water, culturing for 74 hours at the temperature of 36 ℃ in a shaking table with the rotating speed of 200r/min, obtaining a primary seed solution after the culture is finished, and storing the primary seed solution in a refrigerator at the temperature of 4 ℃ for later use;
c. secondary seed liquid culture: under an aseptic environment, inoculating the prepared primary seed liquid into a secondary seed culture liquid containing 2.9% of sucrose, 1.4% of glucose, 1.9% of peptone, 4.4% of calcium carbonate, 1% of yeast extract powder, 0.9% of sodium chloride, 1.7% of beef extract, 0.8% of magnesium sulfate, 0.9% of manganese sulfate, 3.3% of dipotassium hydrogen phosphate and the balance of purified water, and culturing for 73 hours at 35-40 ℃ in a seed tank at the rotating speed of 340r/min to obtain a secondary seed liquid;
d. solid state fermentation: under an aseptic environment, the prepared secondary seed liquid is uniformly inoculated into a solid fermentation culture medium containing 41% of soybean meal, 12% of rice bran, 24% of bran, 14% of calcium carbonate, 2.7% of yeast extract powder, 1.2% of magnesium sulfate, 1.2% of manganese sulfate and the balance of purified water in an inoculation amount of 20% v/v, and fermentation culture is carried out for 70 hours at 37 ℃.
e. Drying and crushing: and after the fermentation is finished, drying the fermentation product at 50 ℃ for 6 hours, and then crushing to 100 meshes to obtain the enterococcus faecium solid state fermentation microbial inoculum.
2) Preparation of probiotic feed additive
1.5 parts of bacillus subtilis, 1.8 parts of bacillus licheniformis, 0.8 part of bacillus pumilus, 1.6 parts of enterococcus faecium solid fermentation microbial inoculum, 5 parts of fructooligosaccharide, 25 parts of kelp powder, 35 parts of zeolite powder and 55 parts of rice bran.
(1) Pulverizing testa oryzae, and sieving with 80 mesh sieve;
(2) mixing the solid fermentation microbial inoculum of bacillus subtilis, bacillus licheniformis, bacillus pumilus and enterococcus faecium with fructooligosaccharide and zeolite powder at the rotation speed of 280r/min for 4min, and mixing uniformly to obtain a mixture;
(3) keeping the rotating speed unchanged, adding the kelp powder and the crushed rice bran into the mixture obtained in the step (2), and stirring for 8min to uniformly mix the kelp powder and the crushed rice bran to obtain the broiler probiotic feed additive.
Example 4
1) Preparing an enterococcus faecium solid fermentation microbial inoculum:
a. screening and domesticating the strains: under the aseptic operation condition, putting enterococcus faecium into an improved MRS culture medium containing 2.6% of glucose, 1.5% of peptone, 1.3% of beef extract, 1.4% of yeast extract powder, 0.8% of sodium chloride, 3.2% of agar powder, 2.6% of calcium carbonate, 1% of diammonium citrate, 800.1% of tween, 0.1% of magnesium sulfate, 0.06% of manganese sulfate and the balance of purified water, streaking, culturing at 37 ℃ for 24 hours, selecting 3-5 typical enterococcus faecium single colonies, streaking into the MRS culture medium respectively, continuously purifying, carrying out passage rejuvenation for 3 times, finally selecting the rejuvenated enterococcus faecium for physiological and biochemical identification, and selecting the optimal enterococcus faecium single colony as a seed strain to serve as a primary seed solution strain after the enterococcus faecium is identified as enterococcus faecium;
b. primary seed liquid culture: under the aseptic operation condition, selecting single colonies of screened and domesticated enterococcus faecium, inoculating the single colonies into a primary seed culture solution which comprises 2.4% of sucrose, 2.1% of glucose, 1.8% of peptone, 5.4% of calcium carbonate, 1.5% of yeast extract powder, 0.8% of sodium chloride, 1.8% of beef extract, 0.8% of magnesium sulfate, 0.8% of manganese sulfate, 3.3% of dipotassium hydrogen phosphate and the balance of purified water, culturing for 72 hours at the temperature of 40 ℃ in a shaking table with the rotating speed of 250r/min, obtaining a primary seed solution after the culture is finished, and storing the primary seed solution in a refrigerator at the temperature of 4 ℃ for later use;
c. secondary seed liquid culture: inoculating the prepared primary seed liquid into a secondary seed culture liquid containing 3.0% of sucrose, 1.5% of glucose, 2.0% of peptone, 4.5% of calcium carbonate, 1% of yeast extract powder, 0.9% of sodium chloride, 1.7% of beef extract, 0.8% of magnesium sulfate, 1.2% of manganese sulfate, 3.3% of dipotassium hydrogen phosphate and the balance of purified water in an inoculation amount of 8% v/v under a sterile environment, and culturing for 74 hours at a temperature of 40 ℃ in a seed tank at a rotating speed of 280r/min to obtain a secondary seed liquid;
d. solid state fermentation: under the aseptic environment, the prepared secondary seed liquid is evenly inoculated into a solid fermentation culture medium containing 40% of soybean meal, 12% of rice bran, 25% of bran, 15% of calcium carbonate, 2.8% of yeast extract powder, 1.2% of magnesium sulfate, 1.2% of manganese sulfate and the balance of purified water, and fermentation culture is carried out for 69 hours at 40 ℃.
e. Drying and crushing: and after the fermentation is finished, drying the fermentation product at 40 ℃ for 5 hours, and then crushing to 90 meshes to obtain the enterococcus faecium solid state fermentation microbial inoculum.
2) Preparation of probiotic feed additive
1.3 parts of bacillus subtilis, 1.4 parts of bacillus licheniformis, 0.6 part of bacillus pumilus, 1.3 parts of enterococcus faecium solid fermentation microbial inoculum, 8 parts of fructooligosaccharide, 30 parts of kelp powder, 25 parts of zeolite powder and 60 parts of rice bran.
(1) Crushing rice bran and sieving the crushed rice bran with a 75-mesh sieve;
(2) mixing the solid fermentation microbial inoculum of bacillus subtilis, bacillus licheniformis, bacillus pumilus and enterococcus faecium with fructooligosaccharide and zeolite powder at the rotation speed of 300r/min for 5min, and mixing uniformly to obtain a mixture;
(3) keeping the rotating speed unchanged, adding the kelp powder and the crushed rice bran into the mixture obtained in the step (2), and stirring for 7min to uniformly mix the kelp powder and the crushed rice bran to obtain the broiler probiotic feed additive.
Comparative example 1
The difference from the embodiment 1 is that the enterococcus faecium solid fermentation inoculum is replaced by lactic acid bacteria, and the other components and the preparation method are the same as the embodiment 1.
Comparative example 2
The difference from example 1 is that enterococcus faecium is directly added without solid state fermentation, and the rest components and preparation method are the same as example 1.
Comparative example 3
The difference from example 1 is that Bacillus licheniformis is replaced by Lactobacillus fermentum, and the other components and preparation method are the same as example 1.
Comparative example 4
The difference from example 1 is that fructo-oligosaccharide is not added, and the rest components and the preparation method are the same as example 1.
Comparative example 5
The difference from the example 1 is that the kelp powder is not added, and the other components and the preparation method are the same as the example 1.
Comparative example 6
The difference from the example 1 is that the zeolite powder is replaced by the talcum powder, and the rest components and the preparation method are the same as the example 1.
Feeding experiment:
and (3) experimental design: 660 three yellow chickens with the same growth state and the same growth period are selected and randomly divided into 11 daily ration treatment groups according to the principle that the weights of the half chickens and the half chickens are consistent, and each group comprises 60 three yellow chickens. Feeding at 34-38 deg.C for 110 days.
Experimental group 1: adding 1.2 wt% of the broiler probiotic feed additive of example 1 to a basal ration;
experimental group 2: adding 0.9 wt% of the broiler chicken prebiotic feed additive of example 2 to a basal ration;
experimental group 3: adding 1.0 wt% of the broiler probiotic feed additive of example 3 to a basal ration;
experimental group 4: adding 1.1 wt% of the broiler probiotic feed additive of example 4 to a basal ration;
experimental group 5: adding 1.2 wt% of the broiler chicken prebiotic feed additive of comparative example 1 to the basic ration;
experimental group 6: adding 1.2 wt% of the broiler chicken prebiotic feed additive of the comparative example 2 to the basic ration;
experimental group 7: adding 1.2 wt% of the broiler chicken prebiotic feed additive of the comparative example 3 into the basic ration;
experimental group 8: adding 1.2 wt% of the broiler chicken prebiotic feed additive of comparative example 4 to the basic ration;
experimental group 9: adding 1.2 wt% of the broiler chicken prebiotic feed additive of the comparative example 5 into the basic ration;
experimental group 10: adding 1.2 wt% of the broiler chicken prebiotic feed additive of the comparative example 6 to the basic ration;
control group: the basic ration is not added with the broiler chicken probiotics feed additive.
After the experiment, the growth conditions and the chicken quality of the three yellow chickens in each group are respectively recorded, and the statistical results are shown in table 1.
TABLE 1
The above-described embodiments are merely illustrative of the preferred embodiments of the present invention, and do not limit the scope of the present invention, and various modifications and improvements of the technical solutions of the present invention can be made by those skilled in the art without departing from the spirit of the present invention, and the technical solutions of the present invention are within the scope of the present invention defined by the claims.
Claims (10)
1. The broiler chicken probiotics feed additive is characterized by comprising the following raw materials in parts by weight:
0.8-1.5 parts of bacillus subtilis, 0.5-1.8 parts of bacillus licheniformis, 0.6-1.2 parts of bacillus pumilus, 1.3-1.8 parts of enterococcus faecium solid fermentation microbial inoculum, 5-8 parts of fructooligosaccharide, 20-30 parts of kelp powder, 25-35 parts of zeolite powder and 50-60 parts of rice bran;
the preparation method of the enterococcus faecium solid fermentation microbial inoculum comprises the following steps:
a. screening and domesticating the strains: under the aseptic operation condition, putting enterococcus faecium into an improved MRS culture medium containing glucose, peptone, beef extract, yeast extract powder, sodium chloride, agar powder, calcium carbonate, diammonium citrate, Tween 80, magnesium sulfate and manganese sulfate, streaking, culturing, selecting typical enterococcus faecium single colonies, streaking into the MRS culture medium respectively, and continuously purifying, subculturing and rejuvenating;
b. primary seed liquid culture: under the aseptic operation condition, inoculating the screened and domesticated enterococcus faecium single colony into a primary seed culture solution containing sucrose, glucose, peptone, calcium carbonate, yeast extract powder, sodium chloride, beef extract, magnesium sulfate, manganese sulfate and dipotassium hydrogen phosphate, carrying out shake culture, obtaining a primary seed solution after finishing the culture, and storing the primary seed solution in a refrigerator at 4 ℃ for later use;
c. secondary seed liquid culture: inoculating the prepared primary seed liquid into a secondary seed culture liquid containing cane sugar, glucose, peptone, calcium carbonate, yeast extract powder, sodium chloride, beef extract, magnesium sulfate, manganese sulfate and dipotassium hydrogen phosphate in a sterile environment, and culturing in a seed tank to obtain a secondary seed liquid;
d. solid state fermentation: uniformly inoculating the prepared secondary seed liquid into a solid fermentation culture medium containing bean pulp, rice bran, wheat bran, calcium carbonate, yeast extract powder, magnesium sulfate and manganese sulfate for fermentation culture in an aseptic environment;
e. drying and crushing: and drying the fermentation product after the fermentation is finished, and crushing to obtain the enterococcus faecium solid fermentation microbial inoculum.
2. The broiler chicken prebiotic feed additive as claimed in claim 1, is characterized by comprising the following raw materials in parts by weight:
1.2 parts of bacillus subtilis, 1.3 parts of bacillus licheniformis, 0.8 part of bacillus pumilus, 1.5 parts of enterococcus faecium solid fermentation microbial inoculum, 6 parts of fructooligosaccharide, 25 parts of kelp powder, 30 parts of zeolite powder and 55 parts of rice bran.
3. The broiler chicken probiotic feed additive of claim 1, wherein in step a, the modified MRS culture medium contains 2.5-2.6% of glucose, 1.3-1.5% of peptone, 1.3% of beef extract, 1.2-1.5% of yeast extract, 0.8% of sodium chloride, 3.1-3.3% of agar powder, 2.5-2.7% of calcium carbonate, 1% of diammonium citrate, 800.1-0.2% of Tween, 0.1% of magnesium sulfate, 0.06% of manganese sulfate and the balance of purified water;
in the step b, the primary seed culture solution contains 2.3-2.4% of sucrose, 2-2.1% of glucose, 1.6-1.8% of peptone, 5.2-5.4% of calcium carbonate, 1.5% of yeast extract powder, 0.8% of sodium chloride, 1.8-2.0% of beef extract, 0.8% of magnesium sulfate, 0.8% of manganese sulfate, 3.3-3.4% of dipotassium hydrogen phosphate and the balance of purified water; in the step b, the temperature of the shaking culture is 35-40 ℃, and the time is 72-74 h.
4. The broiler chicken prebiotic feed additive of claim 1, wherein in step c, the secondary seed nutrient solution contains 2.8-3.0% of sucrose, 1.3-1.5% of glucose, 1.8-2.0% of peptone, 4.3-4.5% of calcium carbonate, 1% of yeast extract powder, 0.9% of sodium chloride, 1.7% of beef extract, 0.8% of magnesium sulfate, 0.8-1.2% of manganese sulfate, 3.3% of dipotassium hydrogen phosphate and the balance of purified water;
in the step c, the inoculation amount of the primary seed liquid is 8-10% of the volume of the secondary seed culture liquid, the temperature of the seed tank culture is 35-40 ℃, and the time is 72-74 h.
5. The broiler chicken prebiotic feed additive of claim 1, wherein in step d, the solid state fermentation culture medium contains 40-43% of soybean meal, 12-13% of rice bran, 23-25% of bran, 11-15% of calcium carbonate, 2.7-2.8% of yeast extract powder, 1.2% of magnesium sulfate, 1.2% of manganese sulfate and the balance of purified water;
in the step d, the inoculation amount of the secondary seed liquid is 15-20% of the volume of the solid fermentation culture medium, the fermentation culture temperature is 35-40 ℃, and the fermentation culture time is 68-72 hours.
6. The broiler chicken probiotics feed additive as claimed in claim 1, wherein in step e, the drying temperature is 30-50 ℃, the drying time is 3-6h, and the mesh number after crushing is 80-100 meshes.
7. A method for preparing a broiler probiotic feed additive as claimed in any one of claims 1-6, characterized in that it comprises the following steps:
(1) pulverizing testa oryzae;
(2) uniformly mixing a solid fermentation microbial inoculum of bacillus subtilis, bacillus licheniformis, bacillus pumilus and enterococcus faecium with fructooligosaccharide and zeolite powder to obtain a mixture;
(3) and (3) adding kelp powder and crushed rice bran into the mixture obtained in the step (2), and uniformly stirring to obtain the prebiotics feed additive.
8. The preparation method of the broiler chicken prebiotic feed additive according to claim 7, wherein the rice bran in step (1) is crushed and sieved with a 60-80 mesh sieve.
9. Use of a broiler probiotic feed additive of any of claims 1-6 in feed.
10. The use of claim 9, wherein the broiler probiotic feed additive is added to the feed in an amount of 0.9-1.2% by weight of the total feed weight.
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