CN116569989A - Biological organic selenium feed additive and preparation method and application thereof - Google Patents
Biological organic selenium feed additive and preparation method and application thereof Download PDFInfo
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- CN116569989A CN116569989A CN202310410975.0A CN202310410975A CN116569989A CN 116569989 A CN116569989 A CN 116569989A CN 202310410975 A CN202310410975 A CN 202310410975A CN 116569989 A CN116569989 A CN 116569989A
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- feed additive
- organic selenium
- bacterial liquid
- biological organic
- bifidobacterium animalis
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- 239000011669 selenium Substances 0.000 title claims abstract description 86
- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 title claims abstract description 85
- 229910052711 selenium Inorganic materials 0.000 title claims abstract description 85
- 239000003674 animal food additive Substances 0.000 title claims abstract description 42
- 238000002360 preparation method Methods 0.000 title claims abstract description 11
- 229940091258 selenium supplement Drugs 0.000 claims abstract description 86
- 240000006024 Lactobacillus plantarum Species 0.000 claims abstract description 32
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims abstract description 32
- 229940072205 lactobacillus plantarum Drugs 0.000 claims abstract description 32
- 244000199866 Lactobacillus casei Species 0.000 claims abstract description 30
- 235000013958 Lactobacillus casei Nutrition 0.000 claims abstract description 30
- 229940017800 lactobacillus casei Drugs 0.000 claims abstract description 30
- 241001134770 Bifidobacterium animalis Species 0.000 claims abstract description 29
- 229940118852 bifidobacterium animalis Drugs 0.000 claims abstract description 29
- 239000000463 material Substances 0.000 claims abstract description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 12
- 244000046052 Phaseolus vulgaris Species 0.000 claims abstract description 6
- 235000010627 Phaseolus vulgaris Nutrition 0.000 claims abstract description 6
- BVTBRVFYZUCAKH-UHFFFAOYSA-L disodium selenite Chemical compound [Na+].[Na+].[O-][Se]([O-])=O BVTBRVFYZUCAKH-UHFFFAOYSA-L 0.000 claims abstract description 6
- 229960001471 sodium selenite Drugs 0.000 claims abstract description 6
- 235000015921 sodium selenite Nutrition 0.000 claims abstract description 6
- 239000011781 sodium selenite Substances 0.000 claims abstract description 6
- 235000015099 wheat brans Nutrition 0.000 claims abstract description 4
- 235000013312 flour Nutrition 0.000 claims abstract description 3
- 230000001580 bacterial effect Effects 0.000 claims description 47
- 239000007788 liquid Substances 0.000 claims description 42
- 238000000855 fermentation Methods 0.000 claims description 25
- 230000004151 fermentation Effects 0.000 claims description 25
- 230000004913 activation Effects 0.000 claims description 15
- 238000000034 method Methods 0.000 claims description 14
- 239000002068 microbial inoculum Substances 0.000 claims description 14
- 238000001035 drying Methods 0.000 claims description 7
- 239000002893 slag Substances 0.000 claims description 6
- 238000002156 mixing Methods 0.000 claims description 5
- 239000008399 tap water Substances 0.000 claims description 5
- 235000020679 tap water Nutrition 0.000 claims description 5
- 241000588769 Proteus <enterobacteria> Species 0.000 claims description 4
- 239000001963 growth medium Substances 0.000 claims description 4
- 238000004519 manufacturing process Methods 0.000 claims description 4
- 239000000203 mixture Substances 0.000 claims description 4
- 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 3
- 239000001888 Peptone Substances 0.000 claims description 3
- 108010080698 Peptones Proteins 0.000 claims description 3
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 3
- 235000015278 beef Nutrition 0.000 claims description 3
- 229940041514 candida albicans extract Drugs 0.000 claims description 3
- 239000012153 distilled water Substances 0.000 claims description 3
- 239000000284 extract Substances 0.000 claims description 3
- 239000008103 glucose Substances 0.000 claims description 3
- 239000002609 medium Substances 0.000 claims description 3
- 235000019319 peptone Nutrition 0.000 claims description 3
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims description 3
- 229920000053 polysorbate 80 Polymers 0.000 claims description 3
- 238000007789 sealing Methods 0.000 claims description 3
- 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 claims description 3
- 239000001393 triammonium citrate Substances 0.000 claims description 3
- 235000011046 triammonium citrate Nutrition 0.000 claims description 3
- 239000012138 yeast extract Substances 0.000 claims description 3
- 238000000108 ultra-filtration Methods 0.000 claims description 2
- 241001465754 Metazoa Species 0.000 abstract description 11
- 239000006041 probiotic Substances 0.000 abstract description 7
- 235000018291 probiotics Nutrition 0.000 abstract description 7
- 239000002699 waste material Substances 0.000 abstract description 6
- 230000009286 beneficial effect Effects 0.000 abstract description 2
- 241000207199 Citrus Species 0.000 description 9
- 235000020971 citrus fruits Nutrition 0.000 description 9
- 230000000052 comparative effect Effects 0.000 description 9
- 230000000694 effects Effects 0.000 description 9
- 239000002131 composite material Substances 0.000 description 7
- 235000013601 eggs Nutrition 0.000 description 7
- 239000003795 chemical substances by application Substances 0.000 description 6
- 239000002054 inoculum Substances 0.000 description 6
- 239000000654 additive Substances 0.000 description 5
- 244000005700 microbiome Species 0.000 description 5
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 4
- 240000008042 Zea mays Species 0.000 description 4
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 4
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 4
- 235000005822 corn Nutrition 0.000 description 4
- 238000011081 inoculation Methods 0.000 description 4
- 244000144972 livestock Species 0.000 description 4
- 230000002195 synergetic effect Effects 0.000 description 4
- 241000287828 Gallus gallus Species 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 230000000968 intestinal effect Effects 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 230000001105 regulatory effect Effects 0.000 description 3
- 235000000346 sugar Nutrition 0.000 description 3
- 230000001502 supplementing effect Effects 0.000 description 3
- 231100000419 toxicity Toxicity 0.000 description 3
- 230000001988 toxicity Effects 0.000 description 3
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 206010012735 Diarrhoea Diseases 0.000 description 2
- 108010070551 Meat Proteins Proteins 0.000 description 2
- 235000019764 Soybean Meal Nutrition 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 230000000996 additive effect Effects 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 125000004122 cyclic group Chemical group 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 230000036039 immunity Effects 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 150000007524 organic acids Chemical class 0.000 description 2
- 235000005985 organic acids Nutrition 0.000 description 2
- 238000009304 pastoral farming Methods 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 239000004455 soybean meal Substances 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 241000282412 Homo Species 0.000 description 1
- 241000282414 Homo sapiens Species 0.000 description 1
- 241000186660 Lactobacillus Species 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 201000009840 acute diarrhea Diseases 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 235000019728 animal nutrition Nutrition 0.000 description 1
- 230000036983 biotransformation Effects 0.000 description 1
- 208000019902 chronic diarrheal disease Diseases 0.000 description 1
- -1 citric acid Chemical class 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 208000016097 disease of metabolism Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- 230000012173 estrus Effects 0.000 description 1
- 231100000086 high toxicity Toxicity 0.000 description 1
- 208000000509 infertility Diseases 0.000 description 1
- 230000036512 infertility Effects 0.000 description 1
- 231100000535 infertility Toxicity 0.000 description 1
- 229940039696 lactobacillus Drugs 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 239000003895 organic fertilizer Substances 0.000 description 1
- 235000019629 palatability Nutrition 0.000 description 1
- 230000008092 positive effect Effects 0.000 description 1
- 244000144977 poultry Species 0.000 description 1
- 238000009374 poultry farming Methods 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000000644 propagated effect Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 230000001850 reproductive effect Effects 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Classifications
-
- 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
- A23K20/30—Oligoelements
-
- 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
- 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
- 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
-
- 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
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
- C12R2001/245—Lactobacillus casei
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
- C12R2001/25—Lactobacillus plantarum
-
- 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 biological organic selenium feed additive and a preparation method and application thereof. The invention mixes the peel residue, bean pulp, puffed bean flour, wheat bran and sodium selenite uniformly, and the material obtained by adjusting the water content is mixed with lactobacillus plantarum, lactobacillus casei and bifidobacterium animalis, fermented, dried and crushed to obtain the biological organic selenium feed additive. The invention not only effectively recycles the agricultural and forestry wastes such as the peel residues, but also can obtain the feed additive rich in the biological organic selenium; in addition, the feed additive also contains rich probiotics, is beneficial to the growth of animals and has better application prospect.
Description
Technical Field
The invention belongs to the technical field of bioengineering, and particularly relates to a biological organic selenium feed additive and a preparation method and application thereof.
Background
With the continuous development of social productivity, the demand of human beings for meat proteins has also increased. However, as a major source of meat proteins, the productivity level of the livestock farming industry is relatively low. Selenium (Se) is one of the microelements necessary for animals to maintain the vitality of the bodies, and the deficiency of the selenium element can lead to slow growth of the animals, cause nutritional and metabolic diseases, and even lead to oestrus disorder, infertility or weakness of newborn piglets after the delivery of sows. The proper selenium supplement in daily ration can not only effectively avoid the problems, but also improve the immunity level of organisms and the reproductive performance of farmed animals, thereby improving the overall productivity of the livestock and poultry farming industry. The inorganic selenium has high toxicity, while the inorganic selenium has higher biocompatibility and lower toxicity, which is the best form for supplementing selenium. At present, most feed production enterprises or farms add sodium selenite or yeast selenium into the feed, wherein the sodium selenite or the yeast selenium has strong toxicity, low absorptivity and certain side effect when used in a large amount; although the use of yeast selenium can solve the problems of toxicity and absorption efficiency, the yeast selenium currently marketed is expensive, which is a considerable expense for large-scale farms. Therefore, there is an urgent need to find an animal selenium supplement that is safe, widely available, inexpensive and easy to absorb.
Disclosure of Invention
The primary aim of the invention is to provide a preparation method of a biological organic selenium feed additive.
The invention also aims at providing the biological organic selenium feed additive obtained by the preparation method.
It is a further object of the present invention to provide the use of the above-described bio-organic selenium feed additive.
The aim of the invention is achieved by the following technical scheme: a preparation method of a biological organic selenium feed additive comprises the following steps:
(1) The peel residue, the bean pulp, the puffed bean flour, the wheat bran and the sodium selenite are mixed according to the mass ratio of 500-800: 100-300: 50-100: 10-30: mixing evenly 0.5-2; adjusting the water content to 50% -70% by using water to obtain a material A;
(2) Uniformly mixing the microbial inoculum and the material A obtained in the step (1) to obtain a fermentation system, and sealing and fermenting; the microbial inoculum comprises lactobacillus plantarum, lactobacillus casei and bifidobacterium animalis;
(3) And (5) drying and crushing after fermentation to obtain the biological organic selenium feed additive.
The mass ratio in the step (1) is preferably 600-700: 200-250: 70-80: 15-25: 0.5 to 1.5; more preferably 650:220:75:20:1.
the peel residue in the step (1) is preferably citrus residue.
The water in step (1) is preferably at least one of tap water and ultrafiltration tap water.
The water content in the step (1) is preferably 60%.
The lactobacillus plantarum, the lactobacillus casei and the bifidobacterium animalis in the microbial inoculum in the step (2) are preferably combined according to the proportion of 1:1-3:1-3 of cell number; more preferably, the combination is carried out according to the ratio of 1:1:1 of the cell number.
The bacterial agent in the step (2) is preferably obtained by combining lactobacillus plantarum bacterial liquid obtained by activation, lactobacillus casei bacterial liquid obtained by activation and bifidobacterium animalis bacterial liquid obtained by activation.
The concentration of the lactobacillus plantarum bacterial liquid obtained by the activation is preferably OD 600 =1.0 to 1.4; more preferably OD 600 ＝1.2。
The concentration of the lactobacillus casei bacterial liquid obtained by the activation is preferably OD 600 =1.0 to 1.4; more preferably OD 600 ＝1.2。
The concentration of the activated bifidobacterium animalis bacterial liquid is preferably OD 600 =1.0 to 1.4; more preferably OD 600 ＝1.2。
The microbial inoculum is preferably prepared by the following steps: inoculating the preserved lactobacillus plantarum, lactobacillus casei and bifidobacterium animalis into an activation culture medium respectively, and then carrying out anaerobic culture to obtain activated lactobacillus plantarum bacterial liquid, lactobacillus casei bacterial liquid and bifidobacterium animalis bacterial liquid;
the composition of the activation medium is as follows: 10g of peptone, 10g of beef extract, 5g of yeast extract, 20g of glucose and K 2 HPO 4 2g，NaOAc 5g，MgSO 4 0.2g，MnSO 4 0.05g, tween-80 1mL, triammonium citrate 2g, distilled water to volume 1L.
The inoculation amount of the inoculation is preferably 0.5-3% (v/v); more preferably 2% (v/v).
The temperature of the anaerobic culture is preferably 28-32 ℃; more preferably 30 ℃.
The addition amount of the microbial inoculum in the step (2) is preferably calculated by adding 0.33-1.00mL of lactobacillus plantarum bacterial liquid, 0.33-1.50mL of lactobacillus casei bacterial liquid and 0.33-1.50mL of bifidobacterium animalis bacterial liquid into every 100g of material A; more preferably, 0.5mL of Lactobacillus plantarum bacterial liquid, 0.5mL of Lactobacillus casei bacterial liquid and 0.5mL of Bifidobacterium animalis bacterial liquid are added to 100g of the material A.
The lactobacillus plantarum is preferably lactobacillus plantarum CCTCC WB 2008944.
The lactobacillus casei is preferably lactobacillus casei CCTCC KB 20081445.
The bifidobacterium animalis is preferably bifidobacterium animalis CGMCC 1.3003.
The sealed fermentation condition in the step (2) is preferably sealed fermentation at 33-39 ℃ for 5-9 days; more preferably, the fermentation is carried out at 33 to 36℃for 5 to 7 days under sealed conditions.
The drying in step (3) is preferably drying at a temperature below 50 ℃; more preferably at 45 ℃.
A biological organic selenium feed additive for regulating animal intestinal flora and improving animal immunity is prepared by the above preparation method.
The application of the biological organic selenium feed additive in preparing feed.
Compared with the prior art, the invention has the following advantages and effects:
(1) The invention provides a preparation process for obtaining an organic selenium feed additive by fermenting pericarp slag through composite probiotics, which can effectively recycle and utilize agricultural and forestry wastes such as pericarp slag, and the agricultural and forestry wastes such as pericarp slag contain a large amount of renewable resources. These renewable resources are not suitable for direct use for humans for a variety of reasons, but these abundant agroforestry wastes are very excellent inexpensive medium substitutes for microorganisms. In the case of citrus pulp, these waste citrus pulp sugar acids are relatively low and have poor palatability and are not suitable for direct consumption, but contain a large amount of organic acids and sugar compounds, mainly citric acid, and microorganisms can directly and rapidly grow using the nutrients therein. In addition, the citrus pulp also contains a great amount of nutritional ingredients such as VC and the like, and the nutritional ingredients are also very beneficial to the cultured animals.
(2) The three probiotics used in the invention are carefully selected and obtained combinations, and firstly, the lactobacillus plantarum and the bifidobacterium animalis are main strains for converting the organic selenium, but the lactobacillus plantarum and the bifidobacterium animalis are only combined and cannot obtain particularly high conversion rate, and the biotransformation of the organic selenium can be further promoted when the lactobacillus casei is added, so that the effect of 1+1+1 is more than 3. Secondly, lactobacillus casei and lactobacillus plantarum can produce a biological preservative, namely the lactobacillus, which greatly improves the preservation period of the product, so that the product is easier to popularize and apply. Finally, the bifidobacterium animalis has very excellent effect of regulating intestinal flora and has very good treatment effect on various acute and chronic diarrhea of livestock and poultry. In conclusion, the three microorganisms in the invention are subjected to synergistic fermentation, so that the excellent animal nutrition selenium supplementing agent rich in organic selenium can be obtained.
(3) The biological organic selenium feed additive can provide safe and easily absorbed selenium element for fed animals, and probiotics contained in the biological organic selenium feed additive can play a role in regulating intestinal flora of the animals. The invention can play an excellent role in the production process of various selenium-enriched livestock products and has better application prospect. The biological organic selenium feed additive provided by the invention has relatively high selenium content in waste discharged from the body after animal absorption, can be used for cultivating and producing selenium-rich agriculture and forestry crops, enables the crops to form efficient cyclic development of selenium-rich agriculture, and provides positive effects for green ecological construction.
Detailed Description
The present invention will be described in further detail with reference to examples, but embodiments of the present invention are not limited thereto.
Example 1
(1) Weighing the following raw materials in parts by mass: 650 parts of citrus pulp (slag obtained after juicing citrus), 220 parts of soybean meal, 75 parts of puffed soybean meal, 20 parts of wheat bran and 1 part of sodium selenite.
(2) And after the raw materials are uniformly mixed, supplementing tap water until the water content is 60%, and obtaining a material A.
(3) Mixing activated lactobacillus plantarum CCTCC WB2008944 bacterial liquid, lactobacillus casei CCTCC KB20081445 bacterial liquid and bifidobacterium animalis CGMCC 1.3003 bacterial liquid according to the volume ratio of 1:1:1 to obtain a composite microbial inoculum; then inoculating the composite microbial inoculum into the material A according to the inoculum size of 1.5% (mL microbial inoculum: g material A) and stirring uniformly again, and then sealing and fermenting at 36 ℃ for 7 days.
Wherein, the activating culture medium comprises the following components: 10g of peptone, 10g of beef extract, 5g of yeast extract, 20g of glucose and K 2 HPO 4 2g，NaOAc 5g，MgSO 4 0.2g，MnSO 4 0.05g, tween-80 1mL, triammonium citrate 2g, distilled water to volume 1L.
Inoculating the preserved Lactobacillus plantarum CCTCC WB2008944, lactobacillus casei CCTCC KB20081445 and Bifidobacterium animalis CGMCC 1.3003 into an activation culture medium respectively at a ratio of 2% (v/v), standing at 30deg.C, and anaerobically culturing until the OD of each bacterial liquid is reached 600 1.2, the activated lactobacillus plantarum CCTCC WB2008944 bacterial liquid, lactobacillus casei CCTCC KB20081445 bacterial liquid and bifidobacterium animalis CGMCC 1.3003 bacterial liquid are obtained.
(4) Drying the fermented material at 45 ℃, and then crushing to obtain the biological organic selenium feed additive.
Example 2
Substantially the same as in example 1, except that the fermentation temperature was 33 ℃.
Example 3
Substantially the same as in example 1, except that the fermentation time was 5 days.
Example 4
Substantially the same as in example 1, except that: the fermentation temperature is 33 ℃ and the fermentation time is 5 days.
Comparative example 1
Substantially the same as in example 1, the only difference is that: the bacterial liquid used was Lactobacillus plantarum CCTCC WB2008944 alone, wherein the inoculum size of Lactobacillus plantarum CCTCC WB2008944 bacterial liquid was 1.5%, and the inoculum size was the same as that of the composite bacterial agent of example 1.
Comparative example 2
Substantially the same as in example 1, the only difference is that: the bacterial liquid used was only lactobacillus casei CCTCC KB20081445, wherein the inoculum size of the lactobacillus casei CCTCC KB20081445 bacterial liquid was 1.5%, which is the same as that of the composite bacterial agent of example 1.
Comparative example 3
Substantially the same as in example 1, the only difference is that: the bacterial liquid used only comprises bifidobacterium animalis CGMCC 1.3003, wherein the inoculation amount of the bacterial liquid of the bifidobacterium animalis CGMCC 1.3003 is 1.5 percent, and the inoculation amount of the bacterial liquid is the same as that of the composite bacterial agent of the embodiment 1.
Comparative example 4
Substantially the same as the examples, the only difference is that: the bacterial liquids used were only lactobacillus plantarum cctccc WB2008944 and lactobacillus casei cctccc KB20081445, wherein the inoculum size of lactobacillus plantarum cctccc WB2008944 and lactobacillus casei cccc KB20081445 was 0.75% respectively, and the sum of the inoculum sizes was the same as that of the composite bacterial agent of example 1.
Comparative example 5
Substantially the same as in example 1, the only difference was that the corn kernels replaced the citrus pulp.
Effect example 1
Selenium content in the additives prepared in examples and comparative examples was measured according to GB/T13883-2008 and the results are shown in Table 1:
TABLE 1 composition of additives in examples
The compositions of the additives prepared in each example and comparative example are shown in Table 1, and by comparing example 1, example 2 and example 3, example 4, the organic selenium content in the feed additive fermented at 36℃is as high as 90.33% (1259.20. Mu.g/g) at 7 days of cultivation time, and this value is only 66.75% (765.93. Mu.g/g) at 33℃of fermentation temperature; also, when the cultivation time was 5 days, the organic selenium content at 36℃fermentation temperature was 64.10% (765.93. Mu.g/g), and only 42.29% (434.02. Mu.g/g) at 33℃conditions, which indicated that the temperature had a very important effect on the conversion of organic selenium. Further, by comparing example 1, example 3 and example 2, example 4, the organic selenium content after 7 days of cultivation was 90.33% (1259.20. Mu.g/g) while only 64.10% (765.93. Mu.g/g) of cultivation was performed for 5 days at a fermentation temperature controlled at 36 ℃; also, when the culture temperature was 33 ℃, the organic selenium content in the additive was 66.75% (787.37. Mu.g/g) in 7 days of fermentation, and only 42.29% (434.02. Mu.g/g) in 5 days of fermentation, which indicated that the organic selenium gradually accumulated in the additive as the fermentation time was prolonged. In addition, comparing the analysis of example 1 with comparative example 5, it was found that the corn kernels were fermented to have relatively low levels of both organic selenium and protein, which may be a significant amount of organic acids and sugars contained in the citrus pulp, which may be a source of fast-acting carbon for microbial growth, while the carbon source in the corn kernels is mainly starch, which is relatively difficult for the microorganisms to utilize, and therefore, after the same period of fermentation, the microorganisms are not propagated at the same rate as in the citrus pulp, and therefore, the converted organic selenium content is relatively low, and the protein content in the feed additive under corn fermentation is relatively low, which may indicate that the microbial content is relatively low.
On the other hand, by comparing example 1 with comparative examples 1, 2 and 3, it can be found that although three probiotics (lactobacillus plantarum CCTCC WB2008944, lactobacillus casei CCTCC KB20081445 and bifidobacterium animalis CGMCC 1.3003) used in the invention can all realize the synthesis of organic selenium under the fermentation condition of single strain, the synthesis efficiency is generally not high, the highest lactobacillus plantarum CCTCC WB2008944 also has the content of organic selenium of 71.20% (835.26 mug/g) under the condition of single strain fermentation, and the three strains are subjected to synergistic fermentation under the same condition, the content of organic selenium can reach 90.33% (1259.20 mug/g), which indicates that the three probiotics possibly have certain synergistic effect, and the conversion efficiency of the organic selenium can be greatly improved when the three probiotics are subjected to the synergistic fermentation.
Effect example 2 raising and detection results
180 healthy 21-week-old laying hens (Guangzhou livestock science institute) are selected and randomly divided into three groups, namely, 60 groups A, 30 groups B and 30 cages. Wherein, group A does not add biological organic selenium feed additive, group B adds 1% (w/w) biological organic selenium feed additive in example 1 in the ration, group C adds 5% biological organic selenium feed additive in example 1 in the ration. The other conditions are all carried out normally according to the regulation system of the chicken farm. The test period is 5 weeks, the pre-test period is 1 week, and the formal test period is 4 weeks. The method for detecting the selenium content in eggs and chicken is carried out according to GB 5009.93-2017.
The invention passes the comparison test, 1% and 5% of the biological organic selenium feed additive in the example 1 are added into the daily ration of the B group and the daily ration of the C group respectively. The selenium content in the eggs of group 3 was detected separately, and it was found that the average selenium content in the eggs produced in group A was 0.115. Mu.g/g, whereas the average selenium content in the eggs of groups B and C was 1.72 and 2.22. Mu.g/g, respectively, significantly higher than the average selenium content in the eggs of group A (p < 0.05). In addition, the influence of the organic selenium feed additive on the laying rate, the average egg weight and the survival rate of the laying hens is not obvious (p is more than 0.05), but the laying hens in the group B can eat all feeds in advance before feeding the laying hens next time, the laying hens in the group C are just opposite, and a part of feeds remain before feeding the laying hens in the group C, so that a small amount of the feed can have the effect of promoting the feeding of the laying hens. Through detection of selenium content of main parts of the whole body of the tested laying hen, the organic selenium feed additive can obviously improve selenium content (p is less than 0.05) of all parts in the laying hen body, and selenium-enriched chicken is formed. On the other hand, the excrement discharged by the selenium-rich laying hens has higher selenium content, can be further prepared into a selenium-rich organic fertilizer, is used for cultivating selenium-rich agriculture and forestry crops, and realizes the efficient cyclic development in the selenium-rich field. The results show that the biological organic selenium feed additive provided by the invention has wide application prospect.
TABLE 2 influence of different proportions of organic selenium feed additives to selenium content in eggs
Note that: ". Times" indicates significant levels of difference between groups, p < 0.05.
TABLE 3 influence of different proportions of organic selenium feed additives to the layer production performance
Note that: "0" indicates that the test layer had consumed the complete feed at normal times, "+" indicates that the test layer had consumed the complete feed before the next feeding, and "-" indicates that the test layer had not consumed the complete feed before the next feeding.
TABLE 4 influence of different proportions of organic selenium feed additives on selenium content in layers
The above examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above examples, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principle of the present invention should be made in the equivalent manner, and the embodiments are included in the protection scope of the present invention.
Claims (10)
1. The preparation method of the biological organic selenium feed additive is characterized by comprising the following steps:
(1) The peel residue, the bean pulp, the puffed bean flour, the wheat bran and the sodium selenite are mixed according to the mass ratio of 500-800: 100-300: 50-100: 10-30: mixing evenly 0.5-2; adjusting the water content to 50% -70% by using water to obtain a material A;
(2) Uniformly mixing the microbial inoculum and the material A obtained in the step (1) to obtain a fermentation system, and sealing and fermenting; the microbial inoculum comprises lactobacillus plantarum, lactobacillus casei and bifidobacterium animalis;
(3) And (5) drying and crushing after fermentation to obtain the biological organic selenium feed additive.
2. The method for preparing the biological organic selenium feed additive according to claim 1, wherein the method comprises the following steps:
the mass ratio in the step (1) is 600-700: 200-250: 70-80: 15-25: 0.5 to 1.5.
3. The method for preparing the biological organic selenium feed additive according to claim 1, wherein the method comprises the following steps:
the peel slag in the step (1) is orange slag;
the water in the step (1) is at least one of tap water and ultrafiltration tap water.
4. The method for preparing the biological organic selenium feed additive according to claim 1, wherein the method comprises the following steps:
the lactobacillus plantarum, the lactobacillus casei and the bifidobacterium animalis in the microbial inoculum in the step (2) are compounded according to the ratio of 1:1-3:1-3 of cell number.
5. The method for preparing the biological organic selenium feed additive according to claim 1, wherein the method comprises the following steps:
the microbial inoculum in the step (2) is obtained by combining lactobacillus plantarum bacterial liquid obtained by activation, lactobacillus casei bacterial liquid obtained by activation and bifidobacterium animalis bacterial liquid obtained by activation;
the concentration of the lactobacillus plantarum bacterial liquid obtained by the activation is OD 600 ＝1.0～1.4；
The concentration of the lactobacillus casei bacterial liquid obtained by the activation is OD 600 ＝1.0～1.4；
The concentration of the activated bifidobacterium animalis bacterial liquid is OD 600 ＝1.0～1.4；
The addition amount of the microbial inoculum in the step (2) is calculated by adding 0.33-1.00mL of lactobacillus plantarum bacterial liquid, 0.33-1.50mL of lactobacillus casei bacterial liquid and 0.33-1.50mL of bifidobacterium animalis bacterial liquid into every 100g of material A.
6. The method for preparing the biological organic selenium feed additive according to claim 5, wherein the method comprises the following steps:
the microbial inoculum is prepared by the following steps: inoculating the preserved lactobacillus plantarum, lactobacillus casei and bifidobacterium animalis into an activation culture medium respectively, and then carrying out anaerobic culture to obtain activated lactobacillus plantarum bacterial liquid, lactobacillus casei bacterial liquid and bifidobacterium animalis bacterial liquid;
the composition of the activation medium is as follows: 10g of peptone, 10g of beef extract, 5g of yeast extract, 20g of glucose and K 2 HPO 4 2g，NaOAc 5g，MgSO 4 0.2g，MnSO 4 0.05g, tween-80 1mL, triammonium citrate 2g, distilled water to volume 1L.
7. The method for preparing the biological organic selenium feed additive according to any one of claims 1 to 6, which is characterized in that:
the lactobacillus plantarum is lactobacillus plantarum CCTCC WB 2008944;
the lactobacillus casei is lactobacillus casei CCTCC KB 20081445;
the bifidobacterium animalis is bifidobacterium animalis CGMCC 1.3003.
8. The method for preparing the biological organic selenium feed additive according to claim 1, wherein the method comprises the following steps:
the sealed fermentation condition in the step (2) is that the sealed fermentation is carried out for 5-9 days at the temperature of 33-39 ℃;
the drying in the step (3) is drying under the condition of less than 50 ℃.
9. A biological organic selenium feed additive is characterized in that: obtained by the production process according to any one of claims 1 to 8.
10. Use of the bioorganic selenium feed additive of claim 9 in the preparation of feed.
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