CN117179177A - Experimental puppy food and preparation method thereof - Google Patents
Experimental puppy food and preparation method thereof Download PDFInfo
- Publication number
- CN117179177A CN117179177A CN202311391117.2A CN202311391117A CN117179177A CN 117179177 A CN117179177 A CN 117179177A CN 202311391117 A CN202311391117 A CN 202311391117A CN 117179177 A CN117179177 A CN 117179177A
- Authority
- CN
- China
- Prior art keywords
- parts
- experimental
- puppy
- food
- probiotics
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 235000013305 food Nutrition 0.000 title claims abstract description 84
- 238000002360 preparation method Methods 0.000 title claims description 26
- 239000006041 probiotic Substances 0.000 claims abstract description 87
- 235000018291 probiotics Nutrition 0.000 claims abstract description 87
- 241000191998 Pediococcus acidilactici Species 0.000 claims abstract description 46
- 239000000843 powder Substances 0.000 claims abstract description 46
- 238000000034 method Methods 0.000 claims abstract description 44
- 235000005911 diet Nutrition 0.000 claims abstract description 30
- 230000037213 diet Effects 0.000 claims abstract description 30
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 29
- 239000000284 extract Substances 0.000 claims abstract description 27
- 230000008569 process Effects 0.000 claims abstract description 21
- 239000003921 oil Substances 0.000 claims abstract description 20
- 235000019198 oils Nutrition 0.000 claims abstract description 20
- 241000272525 Anas platyrhynchos Species 0.000 claims abstract description 16
- 235000013336 milk Nutrition 0.000 claims abstract description 16
- 239000008267 milk Substances 0.000 claims abstract description 16
- 210000004080 milk Anatomy 0.000 claims abstract description 16
- 235000008390 olive oil Nutrition 0.000 claims abstract description 16
- 239000004006 olive oil Substances 0.000 claims abstract description 16
- 241000287828 Gallus gallus Species 0.000 claims abstract description 14
- 235000007164 Oryza sativa Nutrition 0.000 claims abstract description 14
- 240000008042 Zea mays Species 0.000 claims abstract description 14
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims abstract description 14
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims abstract description 14
- 235000005822 corn Nutrition 0.000 claims abstract description 14
- 229910052500 inorganic mineral Inorganic materials 0.000 claims abstract description 14
- 239000011707 mineral Substances 0.000 claims abstract description 14
- 239000000419 plant extract Substances 0.000 claims abstract description 14
- 235000009566 rice Nutrition 0.000 claims abstract description 14
- 239000004575 stone Substances 0.000 claims abstract description 14
- 239000001763 2-hydroxyethyl(trimethyl)azanium Substances 0.000 claims abstract description 13
- 235000019743 Choline chloride Nutrition 0.000 claims abstract description 13
- 235000015278 beef Nutrition 0.000 claims abstract description 13
- SGMZJAMFUVOLNK-UHFFFAOYSA-M choline chloride Chemical compound [Cl-].C[N+](C)(C)CCO SGMZJAMFUVOLNK-UHFFFAOYSA-M 0.000 claims abstract description 13
- 229960003178 choline chloride Drugs 0.000 claims abstract description 13
- 239000000796 flavoring agent Substances 0.000 claims abstract description 13
- 239000011780 sodium chloride Substances 0.000 claims abstract description 13
- 235000019764 Soybean Meal Nutrition 0.000 claims abstract description 12
- 235000019634 flavors Nutrition 0.000 claims abstract description 12
- 239000004455 soybean meal Substances 0.000 claims abstract description 12
- 235000013312 flour Nutrition 0.000 claims abstract description 10
- 210000004185 liver Anatomy 0.000 claims abstract description 10
- 238000005507 spraying Methods 0.000 claims abstract description 9
- 235000012054 meals Nutrition 0.000 claims abstract description 8
- 240000007594 Oryza sativa Species 0.000 claims abstract 4
- 239000000203 mixture Substances 0.000 claims description 33
- 206010012735 Diarrhoea Diseases 0.000 claims description 32
- 230000000529 probiotic effect Effects 0.000 claims description 23
- 244000237330 gutta percha tree Species 0.000 claims description 22
- 241000194032 Enterococcus faecalis Species 0.000 claims description 18
- 241000186604 Lactobacillus reuteri Species 0.000 claims description 18
- 229940032049 enterococcus faecalis Drugs 0.000 claims description 18
- 229940001882 lactobacillus reuteri Drugs 0.000 claims description 18
- 240000006024 Lactobacillus plantarum Species 0.000 claims description 17
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims description 17
- 229940072205 lactobacillus plantarum Drugs 0.000 claims description 17
- 239000003795 chemical substances by application Substances 0.000 claims description 16
- 238000002156 mixing Methods 0.000 claims description 16
- 238000001035 drying Methods 0.000 claims description 12
- 230000001007 puffing effect Effects 0.000 claims description 12
- 238000005496 tempering Methods 0.000 claims description 11
- 239000000463 material Substances 0.000 claims description 10
- 235000020251 goat milk Nutrition 0.000 claims description 9
- NEGYEDYHPHMHGK-UHFFFAOYSA-N para-methoxyamphetamine Chemical compound COC1=CC=C(CC(C)N)C=C1 NEGYEDYHPHMHGK-UHFFFAOYSA-N 0.000 claims description 8
- 239000011782 vitamin Substances 0.000 claims description 7
- 229940088594 vitamin Drugs 0.000 claims description 7
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 abstract description 16
- 238000004321 preservation Methods 0.000 abstract description 9
- 230000000968 intestinal effect Effects 0.000 abstract description 8
- 229910052757 nitrogen Inorganic materials 0.000 abstract description 8
- 230000009286 beneficial effect Effects 0.000 abstract description 6
- 230000000415 inactivating effect Effects 0.000 abstract description 5
- 230000007704 transition Effects 0.000 abstract description 5
- 230000029087 digestion Effects 0.000 abstract description 4
- 230000036737 immune function Effects 0.000 abstract description 4
- 238000004806 packaging method and process Methods 0.000 abstract description 3
- 230000002180 anti-stress Effects 0.000 abstract description 2
- 241000282472 Canis lupus familiaris Species 0.000 description 43
- 241000894006 Bacteria Species 0.000 description 26
- 230000000694 effects Effects 0.000 description 23
- 238000002474 experimental method Methods 0.000 description 22
- 239000002994 raw material Substances 0.000 description 16
- 241000282465 Canis Species 0.000 description 14
- 230000002829 reductive effect Effects 0.000 description 14
- 241001465754 Metazoa Species 0.000 description 13
- 230000001580 bacterial effect Effects 0.000 description 13
- 235000016709 nutrition Nutrition 0.000 description 13
- 210000001035 gastrointestinal tract Anatomy 0.000 description 12
- 238000012360 testing method Methods 0.000 description 12
- 150000001875 compounds Chemical class 0.000 description 11
- 239000007788 liquid Substances 0.000 description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 11
- 241000588724 Escherichia coli Species 0.000 description 10
- 241000209094 Oryza Species 0.000 description 10
- 239000000706 filtrate Substances 0.000 description 9
- 238000005259 measurement Methods 0.000 description 9
- 230000035764 nutrition Effects 0.000 description 9
- 230000004083 survival effect Effects 0.000 description 9
- 241001105894 Murine norovirus Species 0.000 description 8
- 239000003833 bile salt Substances 0.000 description 7
- 238000004140 cleaning Methods 0.000 description 7
- 238000001514 detection method Methods 0.000 description 7
- 239000003814 drug Substances 0.000 description 7
- 239000000835 fiber Substances 0.000 description 7
- 241000607142 Salmonella Species 0.000 description 6
- 239000003242 anti bacterial agent Substances 0.000 description 6
- 229940088710 antibiotic agent Drugs 0.000 description 6
- 210000004027 cell Anatomy 0.000 description 6
- 235000019621 digestibility Nutrition 0.000 description 6
- 230000002550 fecal effect Effects 0.000 description 6
- 238000001914 filtration Methods 0.000 description 6
- 239000001963 growth medium Substances 0.000 description 6
- 238000005457 optimization Methods 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- 239000007787 solid Substances 0.000 description 6
- 229920001817 Agar Polymers 0.000 description 5
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 5
- 239000002253 acid Substances 0.000 description 5
- 239000008272 agar Substances 0.000 description 5
- 230000000844 anti-bacterial effect Effects 0.000 description 5
- 230000008901 benefit Effects 0.000 description 5
- 230000008859 change Effects 0.000 description 5
- 239000002131 composite material Substances 0.000 description 5
- 210000003608 fece Anatomy 0.000 description 5
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 4
- 235000019750 Crude protein Nutrition 0.000 description 4
- 108020004414 DNA Proteins 0.000 description 4
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 description 4
- 238000010171 animal model Methods 0.000 description 4
- 235000019784 crude fat Nutrition 0.000 description 4
- 238000007865 diluting Methods 0.000 description 4
- OVBPIULPVIDEAO-LBPRGKRZSA-N folic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 description 4
- 239000004519 grease Substances 0.000 description 4
- 230000036541 health Effects 0.000 description 4
- 230000036039 immunity Effects 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 239000002068 microbial inoculum Substances 0.000 description 4
- 230000001954 sterilising effect Effects 0.000 description 4
- 238000004659 sterilization and disinfection Methods 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- XOAAWQZATWQOTB-UHFFFAOYSA-N taurine Chemical compound NCCS(O)(=O)=O XOAAWQZATWQOTB-UHFFFAOYSA-N 0.000 description 4
- 230000004584 weight gain Effects 0.000 description 4
- 235000019786 weight gain Nutrition 0.000 description 4
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 3
- 208000020401 Depressive disease Diseases 0.000 description 3
- 244000046052 Phaseolus vulgaris Species 0.000 description 3
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 3
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 3
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 3
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 3
- 230000036528 appetite Effects 0.000 description 3
- 235000019789 appetite Nutrition 0.000 description 3
- 229940093761 bile salts Drugs 0.000 description 3
- 239000011575 calcium Substances 0.000 description 3
- 229910052791 calcium Inorganic materials 0.000 description 3
- 238000007796 conventional method Methods 0.000 description 3
- 238000010790 dilution Methods 0.000 description 3
- 239000012895 dilution Substances 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 238000000855 fermentation Methods 0.000 description 3
- 230000004151 fermentation Effects 0.000 description 3
- 238000011081 inoculation Methods 0.000 description 3
- 238000002955 isolation Methods 0.000 description 3
- 201000003102 mental depression Diseases 0.000 description 3
- 239000011574 phosphorus Substances 0.000 description 3
- 229910052698 phosphorus Inorganic materials 0.000 description 3
- 230000009467 reduction Effects 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 230000002195 synergetic effect Effects 0.000 description 3
- 108020004465 16S ribosomal RNA Proteins 0.000 description 2
- 244000063299 Bacillus subtilis Species 0.000 description 2
- 235000014469 Bacillus subtilis Nutrition 0.000 description 2
- 238000009631 Broth culture Methods 0.000 description 2
- 244000025596 Cassia laevigata Species 0.000 description 2
- 235000006693 Cassia laevigata Nutrition 0.000 description 2
- 206010059866 Drug resistance Diseases 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- 241000282326 Felis catus Species 0.000 description 2
- 241000233866 Fungi Species 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 2
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 2
- 239000004472 Lysine Substances 0.000 description 2
- OVBPIULPVIDEAO-UHFFFAOYSA-N N-Pteroyl-L-glutaminsaeure Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-UHFFFAOYSA-N 0.000 description 2
- 241000191967 Staphylococcus aureus Species 0.000 description 2
- 235000019742 Vitamins premix Nutrition 0.000 description 2
- 230000001142 anti-diarrhea Effects 0.000 description 2
- 239000002956 ash Substances 0.000 description 2
- 229960002685 biotin Drugs 0.000 description 2
- 235000020958 biotin Nutrition 0.000 description 2
- 239000011616 biotin Substances 0.000 description 2
- 230000037396 body weight Effects 0.000 description 2
- 239000000969 carrier Substances 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 229960001231 choline Drugs 0.000 description 2
- OEYIOHPDSNJKLS-UHFFFAOYSA-N choline Chemical compound C[N+](C)(C)CCO OEYIOHPDSNJKLS-UHFFFAOYSA-N 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 230000006866 deterioration Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 239000003085 diluting agent Substances 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 238000001125 extrusion Methods 0.000 description 2
- 229960000304 folic acid Drugs 0.000 description 2
- 235000019152 folic acid Nutrition 0.000 description 2
- 239000011724 folic acid Substances 0.000 description 2
- 210000004211 gastric acid Anatomy 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- 235000013372 meat Nutrition 0.000 description 2
- 229930182817 methionine Natural products 0.000 description 2
- 239000011259 mixed solution Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000000465 moulding Methods 0.000 description 2
- 229960003512 nicotinic acid Drugs 0.000 description 2
- 235000001968 nicotinic acid Nutrition 0.000 description 2
- 239000011664 nicotinic acid Substances 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 235000019629 palatability Nutrition 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 238000003359 percent control normalization Methods 0.000 description 2
- 239000002504 physiological saline solution Substances 0.000 description 2
- 230000002035 prolonged effect Effects 0.000 description 2
- 238000010298 pulverizing process Methods 0.000 description 2
- 229940124513 senna glycoside Drugs 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 238000002791 soaking Methods 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 229960003080 taurine Drugs 0.000 description 2
- -1 thread ends Substances 0.000 description 2
- 239000011573 trace mineral Substances 0.000 description 2
- 235000013619 trace mineral Nutrition 0.000 description 2
- 241000251468 Actinopterygii Species 0.000 description 1
- 206010067484 Adverse reaction Diseases 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 241000193755 Bacillus cereus Species 0.000 description 1
- 241000194108 Bacillus licheniformis Species 0.000 description 1
- 108020000946 Bacterial DNA Proteins 0.000 description 1
- 208000035143 Bacterial infection Diseases 0.000 description 1
- 241000186016 Bifidobacterium bifidum Species 0.000 description 1
- 241000589875 Campylobacter jejuni Species 0.000 description 1
- 229920002261 Corn starch Polymers 0.000 description 1
- 230000004544 DNA amplification Effects 0.000 description 1
- 238000007400 DNA extraction Methods 0.000 description 1
- 208000036649 Dysbacteriosis Diseases 0.000 description 1
- 208000027244 Dysbiosis Diseases 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 206010016952 Food poisoning Diseases 0.000 description 1
- 208000019331 Foodborne disease Diseases 0.000 description 1
- 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 description 1
- 206010020751 Hypersensitivity Diseases 0.000 description 1
- 240000001046 Lactobacillus acidophilus Species 0.000 description 1
- 235000013956 Lactobacillus acidophilus Nutrition 0.000 description 1
- 239000006142 Luria-Bertani Agar Substances 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 241001263478 Norovirus Species 0.000 description 1
- 241000607768 Shigella Species 0.000 description 1
- 108010087230 Sincalide Proteins 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000006838 adverse reaction Effects 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 208000026935 allergic disease Diseases 0.000 description 1
- 230000000433 anti-nutritional effect Effects 0.000 description 1
- 238000002832 anti-viral assay Methods 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- 208000022362 bacterial infectious disease Diseases 0.000 description 1
- 239000003899 bactericide agent Substances 0.000 description 1
- 229910000278 bentonite Inorganic materials 0.000 description 1
- 239000000440 bentonite Substances 0.000 description 1
- SVPXDRXYRYOSEX-UHFFFAOYSA-N bentoquatam Chemical compound O.O=[Si]=O.O=[Al]O[Al]=O SVPXDRXYRYOSEX-UHFFFAOYSA-N 0.000 description 1
- 210000000941 bile Anatomy 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 238000010609 cell counting kit-8 assay Methods 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 239000008120 corn starch Substances 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 230000000875 corresponding effect Effects 0.000 description 1
- 238000010227 cup method (microbiological evaluation) Methods 0.000 description 1
- 230000000120 cytopathologic effect Effects 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 102000038379 digestive enzymes Human genes 0.000 description 1
- 108091007734 digestive enzymes Proteins 0.000 description 1
- 238000007599 discharging Methods 0.000 description 1
- 239000000428 dust Substances 0.000 description 1
- 230000007140 dysbiosis Effects 0.000 description 1
- 239000003925 fat Substances 0.000 description 1
- 235000019197 fats Nutrition 0.000 description 1
- 230000003031 feeding effect Effects 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 235000012041 food component Nutrition 0.000 description 1
- 235000013355 food flavoring agent Nutrition 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 230000002496 gastric effect Effects 0.000 description 1
- 210000004051 gastric juice Anatomy 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 239000000122 growth hormone Substances 0.000 description 1
- 239000007952 growth promoter Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 230000009610 hypersensitivity Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 239000000077 insect repellent Substances 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 229940039695 lactobacillus acidophilus Drugs 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 230000008774 maternal effect Effects 0.000 description 1
- 230000003340 mental effect Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 238000009629 microbiological culture Methods 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 238000013386 optimize process Methods 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 244000045947 parasite Species 0.000 description 1
- 230000000243 photosynthetic effect Effects 0.000 description 1
- 235000021110 pickles Nutrition 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 238000003752 polymerase chain reaction Methods 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000003449 preventive effect Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000004080 punching Methods 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 230000002000 scavenging effect Effects 0.000 description 1
- IZTQOLKUZKXIRV-YRVFCXMDSA-N sincalide Chemical compound C([C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](N)CC(O)=O)C1=CC=C(OS(O)(=O)=O)C=C1 IZTQOLKUZKXIRV-YRVFCXMDSA-N 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- 235000013618 yogurt Nutrition 0.000 description 1
Landscapes
- Fodder In General (AREA)
Abstract
The invention relates to an experimental puppy diet comprising: 10-30 parts of puffed corn, 1-5 parts of puffed soybean meal, 15-30 parts of puffed rice flour, 25-40 parts of chicken meal, 3-10 parts of olive oil, 3-10 parts of duck oil, 0.5-1 part of stone powder, 0.02-0.05 part of choline chloride, 0.1-0.3 part of sodium chloride, 1-4 parts of chicken liver meal, 4-8 parts of beef flavor, 4-8 parts of milk powder and 3-4 parts of multidimensional multi-mineral premix; the method also comprises the step of inactivating probiotics, wherein 1-6g of the inactivating probiotics are added into every kg of puppy food, and the inactivating probiotics comprise Pediococcus acidilactici CGMCC No.23551; it also comprises folium Eucommiae extract, wherein 4-12g folium Eucommiae extract is added into every kg puppy food. The inactivated probiotics, the plant extracts and the like are added into the experimental puppy food, so that the intestinal microecology balance of the experimental puppy is improved, the digestion utilization rate of the puppy food is improved, and the anti-stress capability of the puppy is enhanced; the prepared dog food has low hardness, is easy to eat in the weaning transition period of the puppies, and reduces stress; the milk powder adopts an external spraying process, so that the immune function of experimental puppies is enhanced, and the nitrogen packaging is beneficial to feed preservation.
Description
Technical Field
The invention relates to the technical field of experimental animal feeds, in particular to an experimental puppy food and a preparation method thereof, wherein the experimental puppy food is applied to reducing diarrhea rate of puppies.
Background
In experimental animals, compared with experimental mice, the diarrhea rate of large animals such as experimental dogs, experimental cats and the like is higher, so that the experimental dog feed needs to have the effect of reducing diarrhea and even not diarrhea besides basic nutrition supply. However, because of the animal habit (such as meat nature of cats and omnivorous meat of dogs) and the different nutritional requirements (protein, fat, fiber, calcium, phosphorus, etc.), there are large differences between the feed raw material compositions and the nutritional indexes.
In the feeding process of experimental dogs, weaning diarrhea is a common clinical symptom, the incidence rate is high, serious people even influence the growth and development of dogs and even the post-test result, the incidence causes are multiple, diagnosis and treatment such as viruses, bacteria or parasites are difficult, and diarrhea can be caused by maternal dog causes, stress, food poisoning, nutritional causes and the like. Studies have shown that diarrhea caused by different causes is the same as the destruction of canine intestinal flora. At present, the diarrhea of puppies is mainly prevented by the following ways: 1) Antibiotics: antibiotics have the advantage of quick response and are widely used for treating various diseases, but excessive use of antibiotics can reduce the immunity of animal bodies, easily destroy the balance of micro-ecology in animal intestinal tracts, generate drug resistance after long-term use of antibiotics, and are very unfavorable for disease control in future. 2) Probiotics: diarrhea is correlated with dysbacteriosis, and canine diarrhea is mostly caused by unbalanced intestinal flora due to changes in the word-keeping environment and canine food. At present, four types of probiotics are used for animal cultivation: lactic acid bacteria (such as Lactobacillus reuteri, lactobacillus acidophilus, and Bacillus bifidus), bacillus (such as enterococcus faecalis, bacillus cereus, and Bacillus licheniformis), yeast (such as Saccharomyces cerevisiae and Saccharomyces cerevisiae), and photosynthetic bacteria. 3) Prevention and treatment of traditional Chinese medicine: the traditional Chinese medicine has the characteristics of no medicine residue, difficult generation of drug resistance, safety, wide sources and the like, and is one of the dominant medicines for replacing antibiotics. The treatment measures are feasible in the experimental dog breeding base, but the treatment is not as preventive, and the stress is relieved through the dog food, so that the gastrointestinal health is improved, the weaning diarrhea of the puppies is reduced, the physical quality is ensured to be stable, and the relevant experiments of the later experimental dogs are facilitated.
The standard GB14924 of experimental animal feed prescribes that the experimental animal feed must not be mixed with antibiotics, insect repellents, preservatives, pigments, growth promoters, hormones and other medicines and additives, and has definite requirements (less than or equal to 5 multiplied by 10) on the total number of product colonies 4 CFU/g). Based on the above requirements, the addition of active probiotics to experimental puppies presents a certain infeasibility that increases the number of canine colonies. It is not feasible to add some medicines to the canine food for a long time. For the reasons, the treatment method of diarrhea of experimental puppies is less than that of pet dogs, and the health and stability of animals are emphasized. Therefore, the better method is to reduce stress through dog food and improve the intestinal tract of experimental puppiesHealth and immune function, reducing diarrhea incidence, and ensuring stable physical quality of the later-period experimental dogs. Therefore, there is an urgent need to develop an experimental puppy diet that can reduce diarrhea rate and is nutritionally complete.
Disclosure of Invention
In order to overcome at least one problem in the prior art, the invention provides an experimental puppy food and a preparation method thereof, which optimize the shape and hardness of the puppy food, selectively add inactivated probiotics and plant extracts to the puppy food to improve immunity, and simultaneously spray raw materials such as goat milk powder and the like in vacuum, thereby reducing the loss of built-in vitamins and being easy to keep nutritional components; meanwhile, the working procedures of the experimental dog production enterprises such as goat milk soaking and the like improve the health level of puppies through multiple measures, improve the ration digestibility, reduce the diarrhea rate of the puppies in the feeding process and facilitate the related experiments of later experimental dogs.
In order to achieve the above purpose, the invention adopts the following technical scheme:
a first object of the present invention is to provide an experimental puppy diet comprising, in parts by weight: 10-30 parts of puffed corn, 1-5 parts of puffed soybean meal, 15-30 parts of puffed rice flour, 25-40 parts of chicken meal, 3-10 parts of olive oil, 3-10 parts of duck oil, 0.5-1 part of stone powder, 0.02-0.05 part of choline chloride, 0.1-0..3 part of sodium chloride, 1-4 parts of chicken liver meal, 4-8 parts of beef flavor, 4-8 parts of milk powder and 3-4 parts of multidimensional multi-mineral premix; wherein, the experimental puppy food further comprises inactivated probiotics, and 1-6g of the inactivated probiotics are added into each kg of experimental puppy food.
Further, the experimental puppy diet comprises, in parts by weight: 12-20 parts of puffed corn, 2-4 parts of puffed soybean meal, 18-25 parts of puffed rice flour, 25-32 parts of chicken meal, 4-8 parts of olive oil, 4-8 parts of duck oil, 0.6-0.8 part of stone powder, 0.04-0.05 part of choline chloride, 0.2-0.3 part of sodium chloride, 2-3 parts of chicken liver meal, 4-6 parts of beef flavor, 4-6 parts of milk powder and 3-4 parts of multidimensional multi-mineral premix; 2-4g (preferably 3 g) of inactivated probiotic is added per kg of experimental puppy food.
Further, the inactivated probiotics comprise Pediococcus acidilactici, the Pediococcus acidilactici is COST, the Pediococcus acidilactici is classified and named Pediococcus acidilactici, the preservation number is CGMCC No.23551, the preservation date is 2021 and 10 month 09, the preservation unit is China general microbiological culture Collection center (CGMCC), and the preservation unit address is China center for type 3 microbiological study of national academy of sciences of North Chen West road 1 in the Chat of Beijing.
Furthermore, the inactivated probiotics can also be a compound microbial inoculum composed of Pediococcus acidilactici and other probiotics, for example, the inactivated probiotics also comprise Lactobacillus reuteri, enterococcus faecalis and Lactobacillus plantarum. Specifically, the inactivated probiotics comprise Pediococcus acidilactici, lactobacillus reuteri, enterococcus faecalis and Lactobacillus plantarum, and the Lactobacillus reuteri, the enterococcus faecalis and the Lactobacillus plantarum are respectively preferably: lactobacillus reuteri (trade name: CICC 6226), enterococcus faecalis (trade name: CICC 20419), lactobacillus plantarum (trade name: CICC 6009).
Further, the content ratio of the pediococcus acidilactici, the lactobacillus reuteri, the enterococcus faecalis and the lactobacillus plantarum is 50-200:30-80:2-5:1-3. Preferably, the content ratio is 150-200: 40-60: 2-4:1-2. More preferably, the content ratio is 200:50:3:2. specifically, pediococcus acidilactici content was 2.0X10 10 CFU/g, lactobacillus reuteri content of 5.0X10 9 CFU/g, enterococcus faecalis content of 3.0X10 8 CFU/g, lactobacillus plantarum content of 2.0X10 8 CFU/g。
Further, the experimental puppy food further comprises a plant extract, wherein the plant extract is an eucommia ulmoides leaf extract; 4-12g, preferably 6-10g, more preferably 8g of plant extract is added per kg of experimental puppy food.
Further, the eucommia ulmoides leaf extract is an aqueous extraction and alcohol extraction product, and specifically comprises the following components: cleaning and crushing the collected fresh eucommia ulmoides leaves, adding water for decoction, and filtering out the water to obtain a first filtrate; decocting in water to obtain second filtrate; adding ethanol for suspension, stirring and filtering to obtain a third filtrate; mixing the three filtrates, concentrating, standing, refrigerating, centrifuging to obtain paste, drying under reduced pressure, and pulverizing into powder.
Further, the multi-dimensional multi-ore premix consists of 1% of compound vitamin premix and 1%.5% compound trace element premix, 1% lysine, 2% taurine, 5% methionine and carrier powder, and mainly contains VE 1200-1800IU/kg, VB 1 300-420mg/kg、VB 2 200-300mg/kg, nicotinic acid 1000-1500mg/kg, biotin 2-6mg/kg, folic acid 30-60mg/kg, choline 300-600mg/kg, VB 6 100-300mg/kg、VB 12 500-700mg/kg, VA200000-400000IU/kg, VD60000-80000IU/kg, fe 1.5-3.0g/kg, cu 25-50mg/kg, mn 500-800mg/kg, zn 800-1200mg/kg, se 0.8-2.0mg/kg. It is understood that other premixes conventionally used in the art may be employed for the multi-dimensional multi-mineral premix.
In one embodiment, the experimental puppy diet comprises: 15 parts of puffed corn, 3 parts of puffed soybean meal, 20.55 parts of puffed rice flour, 30 parts of chicken powder, 7 parts of olive oil, 7 parts of duck oil, 0.8 part of stone powder, 0.05 part of choline chloride, 0.3 part of sodium chloride, 2 parts of chicken liver powder, 5 parts of beef flavor agent, 5 parts of milk powder and 4 parts of multi-vitamin multi-mineral premix; wherein 3g of inactivated probiotic bacteria (Pediococcus acidilactici) is added to each kg of experimental puppy food.
In one embodiment, the experimental puppy diet comprises: 15 parts of puffed corn, 3 parts of puffed soybean meal, 20.55 parts of puffed rice flour, 30 parts of chicken powder, 7 parts of olive oil, 7 parts of duck oil, 0.8 part of stone powder, 0.05 part of choline chloride, 0.3 part of sodium chloride, 2 parts of chicken liver powder, 5 parts of beef flavor agent, 5 parts of milk powder and 4 parts of multi-vitamin multi-mineral premix; wherein, 8g of eucommia ulmoides leaf extract and 3g of inactivated probiotics (Pediococcus acidilactici) are added into each kg of experimental puppy food.
In one embodiment, the experimental puppy diet comprises: 15 parts of puffed corn, 3 parts of puffed soybean meal, 20.55 parts of puffed rice flour, 30 parts of chicken powder, 7 parts of olive oil, 7 parts of duck oil, 0.8 part of stone powder, 0.05 part of choline chloride, 0.3 part of sodium chloride, 2 parts of chicken liver powder, 5 parts of beef flavor agent, 5 parts of milk powder and 4 parts of multi-vitamin multi-mineral premix; wherein, 8g of eucommia ulmoides leaf extract and 3g of inactivated probiotics (composed of Pediococcus acidilactici, lactobacillus reuteri, enterococcus faecalis and Lactobacillus plantarum with the content ratio of 200:50:3:2) are added into each kg of experimental puppy food.
In a second aspect, the invention provides a method of preparing the experimental puppy diet of any one of the first aspects of the invention, comprising the steps of:
s1) sequentially carrying out primary mixing, primary crushing, pre-curing and secondary crushing on corn, soybean meal and rice in predetermined parts by weight to obtain a first mixture;
s2) mixing the first mixture with chicken powder, stone powder, sodium chloride, choline chloride, multidimensional multi-mineral premix and inactivated probiotics for the second time according to the preset weight parts to obtain a second mixture, and adding the second mixture in the step if the second mixture contains plant extracts;
s3) tempering the second mixture, adding predetermined parts by weight of duck oil and olive oil in the tempering process, puffing, granulating and drying to obtain a third mixture;
and S4) spraying the third mixture by using the goat milk powder and the beef flavor agent in predetermined weight parts to obtain the experimental puppy food.
Further, the step S3 specifically includes: tempering the second mixture at 130 ℃ under 0.5Mpa steam, and adding mixed oil of duck oil and olive oil in the tempering process; puffing and granulating after the material temperature reaches 92 ℃, and drying at 120 ℃ for 10min and 60 ℃ for 10min after forming, wherein the diameter of a circular die hole is 7 mm. The internal grease is used for reducing the hardness of the dog food, and the dog food is not required to be soaked and softened and fed to puppies.
Further, the dog food after the spraying step is cooled and then packed by nitrogen.
In a third aspect, the invention provides the use of a laboratory puppy diet as described in any one of the first aspects of the invention or a method of preparation as described in any one of the second aspects of the invention for reducing the rate of diarrhea in a laboratory puppy.
Further, in the application, the experimental dog food is fed 2 times a day, and the feeding amount is 2.5% -3.5% of the weight of the experimental dog.
Compared with the prior art, the invention has the following beneficial effects by adopting the technical scheme:
the experimental puppy food prepared by the method meets the requirements of GB14924.1 and GB14924.2, and the weaning stress of the experimental puppy is reduced, the immune function and intestinal microecology balance of the experimental puppy are improved, the incidence of diarrhea of the experimental puppy in the feeding process is reduced, and the body quality of the experimental puppy is ensured to be stable through the optimization of the nutrition composition of the feed and the optimization of the preparation process.
The inactivated probiotics (single or compound) are added into the experimental puppy food, and have the following advantages: 1) The safety is high: the use of a large amount of live probiotics can possibly cause the problems of bacterial infection, harmful metabolites, hypersensitivity and the like, compared with the inactivated probiotics, the inactivated probiotics are in a more stable state due to the activity loss, and the live probiotics cannot grow and propagate in a host body, so that the live probiotics have higher safety; 2) Stability is high: the living probiotics have strict requirements on preservation and transportation conditions and shelf life, most of the living probiotics in the market at present need to be transported and stored at low temperature (4 ℃), and the shelf life is only several days to more than ten days. On the other hand, the medicine enters the digestive tract, is easily influenced by gastric acid, bile, digestive enzymes and the like, and cannot reach the intestinal tract to play a role. There is no requirement to compare the inactivated probiotic preparation; 3) Convenience: the viable bacteria preparation usually needs to reach a certain number of viable bacteria to exert curative effect, compensates the consumption of viable bacteria in the process of passing through the gastrointestinal tract, and can provide enough viable cells for colonization in the intestinal tract of a host. However, if the probiotic preparation is excessively added, adverse reactions can be caused, so that the use amount of the probiotic live bacteria preparation is difficult to control, and the inactivated probiotic preparation has no limit on the number of bacteria in the use process.
The plant extract, namely the eucommia ulmoides leaf extract, is added into the experimental puppy food, and has the effects of resisting oxidation, regulating immunity, inhibiting bacteria in a broad spectrum, scavenging free radicals, improving immunity and the like, and can relieve diarrhea of animal population caused by stress and resistance drop to a certain extent.
The invention adopts the puffing raw materials, carries out the advanced curing treatment on the cereal raw materials such as corn, bean pulp, rice and the like, reduces the anti-nutritional factors, is beneficial to the nutrition absorption, and specifically comprises the following steps: 1) The digestibility is high: the raw materials are pretreated by adopting a puffing process, so that cell walls in the fiber structure of the raw materials can be destroyed and softened, the gelatinization degree of starch is improved, and the digestion utilization rate of feed is improved; 2) The total number of colonies is small: harmful bacteria such as salmonella, escherichia coli and the like generated in the puffing process are killed completely, the total colony count of the raw materials is reduced, and the dog food sanitation index is greatly improved; 3) The puffing degree is good: the puffed raw materials are used for further improving the puffing degree of the dog food, improving the crispness and improving the palatability of the dog food.
The preparation method has the advantages that the hardness of the puppy food is low, the feeding transition of the puppy is facilitated, the hardness of the puppy food is reduced by adopting a process mode of built-in grease (duck oil and olive oil), meanwhile, the adhesion of built-in grease in a mixer is avoided, the hardness of the puppy food is reduced by built-in grease, the puppy food can be crushed slightly with great effort, the transition from liquid to solid in the weaning period is facilitated, and the feeding stress is reduced; in terms of process equipment, the upper die holes of the die are staggered to prevent adhesion during discharging, a dehumidifier is arranged from the discharge of the bulking machine port to the baking oven section, and the storage time of the puppy food is prolonged.
According to the invention, after the materials are dried, the goat milk powder is externally sprayed, nutrition is provided by the milk powder, the weaning stress of the puppy is relieved, meanwhile, the influence of high-temperature and high-pressure deterioration of the milk powder on nutrition absorption can be avoided by the externally sprayed milk powder, the nutrition loss and the easy deterioration of the milk powder in high temperature and high pressure can be avoided by the measures, and part of the goat milk powder can be sucked into the food of the puppy and is partially stuck to the surface by vacuum negative pressure spraying, so that the smell phagostimulation is improved.
The puppy food is packaged by adopting nitrogen, so that extrusion deformation of the puppy food is effectively prevented, the freshness of the puppy food is maintained, and the palatability is improved; the nitrogen package has certain fresh-keeping and extrusion-preventing functions, and extrusion denaturation in the transportation and storage processes is prevented due to the low hardness of the puppy food.
The experimental puppy food disclosed by the invention has the advantages that the nutrition of the adopted raw materials is comprehensive and balanced, the safety is high, the experimental puppy food is easy to digest and absorb, in particular, inactivated probiotics, plant extracts and the like are added, so that the intestinal microecology balance of an experimental puppy is improved, the digestion utilization rate of the puppy food is improved, the anti-stress capability of the puppy is enhanced, meanwhile, the hardness of the puppy food is low, the puppy is easy to eat in the weaning transition period, the transition from liquid to solid is avoided, and the stress is reduced; in addition, the external spraying process of milk powder is added, the process of soaking the dog food and the goat milk in the production enterprises is reduced, the manpower input is reduced, the immune function of the experimental puppies is enhanced, the body health of the experimental dogs is maintained, and the nitrogen package is beneficial to the fresh keeping of the feed and the service life of the feed is prolonged.
Detailed Description
The following description of the technical solutions in the embodiments of the present invention will be clear and complete, and it is obvious that the described embodiments are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention. The experimental procedures, which are not specified in the following examples, are generally determined according to national standards. The experimental materials not shown in the examples below are all commercially available. The equipment used in each step in the following examples is conventional equipment. If the corresponding national standard does not exist, the method is carried out according to the general international standard, the conventional condition or the condition recommended by the manufacturer. Unless otherwise indicated, all parts are parts by weight and all percentages are percentages by mass. Unless defined or otherwise indicated, 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. In addition, any method and material similar or equivalent to those described may be used in the methods of the present invention.
It should be noted that, without conflict, the embodiments of the present invention and features of the embodiments may be combined with each other. The invention will be further illustrated, but is not limited, by the following examples.
EXAMPLE 1 isolation, identification and Properties of Pediococcus acidilactici
In this example, a strain of Pediococcus acidilactici was isolated from canine feces, and the isolation and identification steps included:
1. isolation of strains
(1) Stool sample test methods reference the handbook for the identification of Burjie bacteria (ninth edition), the specific procedures are as follows: weighing 0.5g of fecal sample, mixing with 5mL of sterile physiological saline, and filling with vortex oscillatorMixing under vibration, continuously diluting 1mL of the mixed solution in sterile EP tube by ten times, and respectively sucking 100 μL of the mixed solution with a dilution gradient of 10 4 、10 5 、10 6 The dilutions were spread evenly onto the surface of MRS agar plates. (2) The collected water at the bottom of the fish tank is 5mL, and 100 mu L of the water is directly absorbed after being fully and uniformly mixed and uniformly coated on the surface of an MRS agar plate. (3) Cleaning the surface of the collected hand pickle with 10mL of sterile normal saline, retaining the normal saline after cleaning, fully mixing, continuously diluting with ten times of the normal saline, and sucking 100 mu L of diluted solution with a gradient of 10 4 Evenly spread on the surface of MRS agar plate. (4) Diluting the collected 1 drop of manual yoghurt by using 1mL of sterile physiological saline, fully mixing, diluting three times by ten times, respectively sucking 100 mu L of diluents with different dilution gradients, and uniformly coating the diluents on the surface of an MRS agar plate.
After the coated MRS culture medium is subjected to standing culture for 48 hours in a 37 ℃ incubator, single colonies with different morphological sizes are selected and inoculated into 10mL MRS broth culture medium, and marked. After standing culture for 24 hours at 37 ℃, a sterile inoculating loop is dipped with bacterial liquid to carry out MRS agar plate streak inoculation purification. Repeating the above steps for three times to obtain the purified strain.
2. Identification of strains
Inoculating the strain into MRS broth culture medium according to 4%, standing at 37deg.C for culturing for 24 hr, shaking and mixing the bacterial liquid, sucking 1mL bacterial suspension into EP tube, centrifuging at 5000r/min for 10min, decanting supernatant, repeating for three times, extracting isolate DNA according to bacterial DNA extraction kit operation instruction, detecting the concentration and purity of the extracted sample, and storing at-80deg.C. The resulting isolate DNA was subjected to the polymerase chain reaction using 16S rDNA universal primers (pollymerase chain reaction, PCR). Wherein the PCR reaction system (25. Mu.L) comprises: ddH 2 O9. Mu.L, 2X HieffRobust PCR Master Mix (With Dye) 12.5. Mu. L, primer forward (10. Mu.M) 1.25. Mu. L, primer reverse (10. Mu.M) 1.25. Mu.L, bacterial genomic DNA 1.0. Mu.L. The PCR cycle parameters were: 94 ℃ for 4min;94℃for 10s, 55℃for 20s, 72℃for 30s,34 cycles; and at 72℃for 5min. The primer sequences used were: upstream primer 27F (AGAGTTTGATCCTGGCTCAG), downstream primer 1492R (GGTTACCTTTGTTACGACTT).
Taking the DNA amplified product of the isolated strain to carry out 1% agarose gel electrophoresis test for detection, observing the size of a band and the purity of a sample, wherein the amplified target fragment is about 1500bp; and sending the obtained product to a biological engineering (Shanghai) stock limited company, sequencing a DNA amplification product, comparing the sequence of the 16S rDNA sequence obtained by sequencing with related sequences in a GenBank database, performing multiple comparison by using Blast program, identifying the separated strain as Pediococcus acidilactici COST, and preserving the separated strain with the preservation number of CGMCC No.23551.
3. Acid and bile salt resistance, high temperature resistance, bacteriostasis and antiviral ability
Acid and bile salt resistance determination: under aseptic condition, 100 μl of Pediococcus acidilactici COST was inoculated into MRS broth with pH of 1.5, 2.0 and pH3.5, respectively, 0.1%, 1%, 2% and 3% bile salts, and the initial number of viable Pediococcus acidilactici COST was 1×10 8 CFU/mL, cultured at constant temperature in an incubator at 37 ℃. The coated plates were sampled at 0h and 2h for counting viable bacteria, and the survival rate was calculated. The results show that: when the pH value is 2 (the pH value in canine gastric juice), the survival rate of the pediococcus acidilactici COST after 2 hours of culture is about 89%, which shows that the strain COST has higher acid resistance, can resist gastric acid and can smoothly reach intestinal tracts to play a role; pediococcus acidilactici COST has a survival rate of about 73% after 2 hours treatment in a culture medium containing 0.1%, 1%, 2% and 3% of bile salts, and shows that the strain COST has high bile salt resistance, can resist the bile salts in intestinal juice, and can preserve vitality and play a role.
High temperature resistance measurement: and (3) taking 10mL of the pediococcus acidilactici COST fermentation liquor, treating the fermentation liquor in a test tube in a water bath kettle at 85 ℃ for 30min, respectively detecting the activity (viable count) before and after treatment, and calculating the survival rate. The results show that: the survival rate of the pediococcus acidilactici COST reaches 85 percent, and the pediococcus acidilactici COST has stronger high temperature resistance.
And (3) bacteriostasis measurement: the viable count is 10 6 CFU/mL of escherichia coli, bacillus subtilis, staphylococcus aureus and salmonella are respectively and uniformly mixed with LB agar medium according to the inoculation amount of 10mL/L, punching is carried out by adopting an oxford cup method, and pediococcus acidilactici COST 10 is taken 8 CFU/mL 00 μL were respectively put into the wells, allowed to stand and spread at 4℃for 2 hours, then cultured at 37℃for 24 hours, and the inhibition thereof was measuredThe diameter of the fungus circle is used as a control by using the MRS culture medium without fungus inoculation. The results show that: the pediococcus acidilactici COST has antibacterial effect (the diameter of a bacteriostasis circle is not less than 13 mm) on escherichia coli, bacillus subtilis, staphylococcus aureus and salmonella, and has strong antibacterial effect on the escherichia coli and the salmonella.
Antiviral assay: murine Norovirus (MNV) infection systems are employed. The mouse mononuclear macrophage RAW264.7 cells were placed in DMEM medium containing 10% fbs, counted, and sub-packed into T25 cell culture flasks, and the medium was replenished for culture, and RAW264.7 cells (10 4 Well) was inoculated into a culture plate into which 100. Mu.L of DMEM medium containing 2% FBS was added per well. Pediococcus acidilactici COST fermentation broth (10) 8 CFU/mL) and MNV virus solution in a volume ratio of 1:1, mixing the components according to a proportion, incubating for a certain time at room temperature, inoculating MNV virus liquid (100 mu L), MNV virus liquid+active Pediococcus acidilactici COST (100 mu L) and MNV virus liquid+inactive Pediococcus acidilactici COST (100 mu L) to culture plates respectively, continuously observing for 7 days after cytopathic effect occurs, and calculating the cell survival rate by adopting a cell counting reagent (CCK-8) method. The results show that: compared with an MNV virus liquid control group (the survival rate is about 12%), the survival rate of RAW264.7 cells after being infected by MNV is improved (about 21%) by adopting the inactivated Pediococcus acidilactici COST treatment, namely the method has a certain capability of reducing the infection power of norovirus, but the effect is inferior to that of active Pediococcus acidilactici COST (the survival rate is about 28%), and compared with active Pediococcus acidilactici, the inactivated probiotic is in a more stable state due to the loss of activity, so the method has higher safety and is more suitable for experimental puppies.
Example 2 preparation of experimental puppy diet
This example is a certain preferred experimental puppy diet and its preparation, the composition of each experimental puppy diet is shown in the following table:
in the experimental puppy food, the multidimensional multi-mineral premix consists of 1% of compound vitamin premix, 1.5% of compound trace element premix, 1% of lysine, 2% of taurine, 5% of methionine and carrier secondary powder, and mainly comprises VE 1600IU/kg, VB 1 380mg/kg、VB 2 230mg/kg, nicotinic acid 1200mg/kg, biotin 4mg/kg, folic acid 50mg/kg, choline 500mg/kg and VB 6 200mg/kg、VB 12 595mg/kg, VA300000IU/kg, VD72000IU/kg, fe 2.3g/kg, cu 35mg/kg, mn 655mg/kg, zn 1000mg/kg, se 1.4mg/kg.
A preferred process for preparing the experimental puppy diet described above comprises the steps of:
(1) Preparation of inactivated probiotics
Selection of strains: pediococcus acidilactici (preservation number CGMCC No. 23551) has certain antibacterial capability on escherichia coli, salmonella, shigella, campylobacter jejuni and the like, and has good acid resistance, bile salt resistance and stress resistance; lactobacillus reuteri (commodity number: CICC 6226), enterococcus faecalis (commodity number: CICC 20419), lactobacillus plantarum (commodity number: CICC 6009) all have excellent acid resistance, antibacterial ability and stress resistance, and the strains are suitable for preparing inactivated probiotics and are added into animal feed;
preparing bacterial liquid: respectively picking 3wt% of Pediococcus acidilactici single colony, enterococcus faecalis single colony, lactobacillus plantarum single colony and Lactobacillus reuteri single colony, inoculating the single colony into a culture medium containing 2.5% of corn starch, 3% of bean pulp and 2% of glucose, culturing at an initial pH of 5.5 and a liquid loading amount of 30mL at 37 ℃ for 12 hours, and adopting a colony plate counting method to adjust the concentration of bacterial liquid;
preparation of solid probiotics: bentonite and mannooligosaccharide are used as drying carriers, and the carriers and bacterial suspension are mixed according to a ratio of 2:3, after mixing in proportion, carrying out blast drying at 50 ℃ for 6 hours to obtain a solid probiotic bacterial agent;
preparation of solid-state inactivated probiotics: inactivating the solid probiotic bacteria agent through CO60 irradiation of 45kGy to obtain the solid inactivated probiotic bacteria agent.
(2) Preparation of eucommia ulmoides leaf extract
Cleaning and crushing the collected fresh eucommia ulmoides leaves, adding 8 times of water, decocting for 20min, and filtering out the water to obtain a first filtrate; adding 4 times of water, decocting for 30min, filtering out water to obtain second filtrate; adding 3 times of ethanol for suspension, adjusting pH to 3, stirring, filtering, adjusting pH of the filtrate to neutrality, and filtering to obtain third filtrate; mixing the three filtrates, concentrating, standing, refrigerating, centrifuging at 2000r/min for 35min to obtain paste, drying under reduced pressure, and pulverizing into powder.
(3) And (3) preparing feed: the production process of the experimental dog food mainly comprises the working procedures of raw material receiving and cleaning, coarse crushing, batching, mixing, expanding, secondary crushing, secondary mixing, tempering, puffing, drying, spraying, cooling, conveying, grading, cleaning, finished product packaging and the like, and adopts the processes of crushing firstly and then matching.
Before the feed raw material is received, sampling quality inspection is carried out, and the nutrition content, color and taste of the feed are in accordance with acceptance standards, and the feed is free from mildew. The material is lifted to a raw material bin through a hopper type lifting machine by a material feeding port, and impurities such as thread ends, dust, broken stone, scrap iron and the like are removed by a primary cleaning sieve and a permanent magnet cylinder before entering the raw material bin. It will be appreciated that the total amount of feed prepared may be adjusted according to the capacity of the actual plant, but that the weight fraction of each component is unchanged.
The corn, the bean pulp and the rice which are automatically weighed are transferred into an initial mixer by a lifting machine to be mixed for 5 minutes, and then transferred into a bin to be crushed to prepare for crushing. The particle size of the crushed material particles is 0.7-0.8mm, and the first mixture is obtained. The first mixture is sent into a bulking machine for pre-curing, the bulking machine is provided with a pair of screws and screw sleeves, after the second mixture enters a bulking cavity, the second mixture is extruded, rubbed, sheared and the like between the screw sleeves, the internal pressure is continuously increased to 4MPA at maximum, and the temperature is continuously increased to 140 ℃. In the period of 3-7 minutes, the temperature and the pressure are rapidly increased, the tissue structure of the second mixture is changed, and coarse fibers are destroyed to kill harmful bacteria such as salmonella. The high-temperature and high-pressure mixture is discharged from the discharge hole, the pressure is suddenly released, the water release part is flashed, and the cooled material is in a loose and porous structure. The whole pre-curing process was carried out for 10 minutes.
Secondary crushing the cured raw materials, adding the rest raw material components including chicken powder compound premix, stone powder, sodium chloride, choline chloride, inactivated probiotics and eucommia ulmoides leaf extract, and carrying out secondary mixing, and mixing for 2.5 min to obtain a second mixture; tempering the second mixture at 130 ℃ under 0.5Mpa steam, and adding the mixed oil of the duck oil and the olive oil in the tempering process. Puffing and granulating after the temperature reaches 92 ℃, forming, drying at 120 ℃ for 10min and 60 ℃ for 10min, spraying with goat milk powder and beef flavoring agent, cooling, and packaging with nitrogen.
The ratio content of probiotics contained in the prepared experimental puppy food can be adjusted according to actual conditions, for example, the content ratio of Pediococcus acidilactici, lactobacillus reuteri, enterococcus faecalis and Lactobacillus plantarum is 50-200:30-80:2 to 5:1 to 3. Preferably, the inactivated probiotic bacteria of each puppy diet in the above table have a Pediococcus acidilactici content of 2.0X10 10 CFU/g, lactobacillus reuteri content of 5.0X10 9 CFU/g, enterococcus faecalis content of 3.0X10 8 CFU/g, lactobacillus plantarum content of 2.0X10 8 CFU/g。
Example 3-performance verification of experimental puppy diet with inactivated probiotic added
In this example, performance of the canine food prepared by adding inactivated probiotics and optimizing the preparation process was verified, and the puppy food 1 prepared in example 2 were fed to beagle dogs, i.e., a control group and an experimental group, respectively. The beagle dogs were evaluated for growth curve, apparent nutrient digestibility, fecal formation, and mental status during feeding.
Control group 1 (prepared without eucommia ulmoides leaf extract and inactivated probiotics, example 2 method): the feed (15 parts of puffed corn, 3 parts of puffed soybean meal, 20.55 parts of puffed rice flour, 30 parts of chicken powder, 7 parts of olive oil, 7 parts of duck oil, 0.8 part of stone powder, 0.05 part of choline chloride, 0.3 part of sodium chloride, 2 parts of chicken liver powder, 5 parts of beef flavor, 5 parts of milk powder and 4 parts of multi-vitamin multi-mineral premix) prepared by adopting the preparation method of the embodiment 2 does not contain eucommia ulmoides leaf extract and inactivated probiotics;
control group 2 (without eucommia ulmoides leaf extract, with inactivated probiotics, prepared by conventional method): the feed composition is the same as that of the puppy food 1, and the puppy food (containing inactivated probiotics (Pediococcus acidilactici), and not using puffing raw materials, oil external spraying, goat milk powder/VB and other built-in materials) prepared by a conventional method;
experimental group 1 (puppy diet 1 prepared in example 2): feed without folium Eucommiae extract and with inactivated probiotics (Pediococcus acidilactici);
experimental group 2 (puppy diet 2 prepared in example 2): feed without folium Eucommiae extract and inactivated probiotics (four probiotic compound bacteria).
The dry matter, crude protein, crude fiber, crude fat, crude ash, calcium and phosphorus were substantially identical for the five groups of canine foods.
TABLE 1 Table of nutritional ingredients for beagle puppies
| Project | Control group 1 | Control group 2 | Experiment group 1 | Experiment group 2 |
| Dry matter, percent | 90.0 | 90.7 | 91.8 | 92.8 |
| Crude protein, percent | 30.8 | 30.5 | 30.9 | 31.0 |
| Coarse fibers, percent | 2.3 | 2.4 | 2.4 | 2.3 |
| Crude fat,% | 19.8 | 20.5 | 20.6 | 20.3 |
| Coarse ash, percent | 5.2 | 5.3 | 5.3 | 5.2 |
| Calcium, percent | 1.25 | 1.27 | 1.28 | 1.29 |
| Phosphorus,% | 1.01 | 0.98 | 1.03 | 1.05 |
Grouping and feeding animals: the influence of the experimental puppies on the growth and apparent digestibility of the weaned beagle dogs was evaluated by feeding the experimental puppies with the conventional diet. The beagle dogs each had 12 animals (6 animals for male and female animals) (initial body weight of 2.3-2.6 kg) per feed, 1 animal per cage, and the test period was 2 months (2 months old-4 months old). The puppies were fed once in the morning and afternoon during the period, and the feeding amount was adjusted according to 2.5% -3.5% of the visual weight.
Method and main instrument for detection: crude protein measurement, nitrogen determination method and Kjeldahl nitrogen determination instrument; crude fiber measurement, drying method and crude fiber measurement instrument; crude fat determination, drying method, crude law enforcement determination instrument. The detection method is carried out according to the relevant standard of feed detection in national standards. Wherein: measurement of crude protein in feed GB/T6432-1994, measurement of crude fat in feed GB/T6433-1994.
1) Feed nutrition apparent digestibility (at the end of the experiment, 4 months old)
Table 2-nutritional digestibility of beagle puppies
In the experiment for 2 months, compared with the feed control group 2 prepared by the conventional method, the puppy has the advantages that the puppy is more convenient to eat and chew, has better taste and has better digestion and utilization effects on the canine foods (experiment group 1 and experiment group 2) prepared by the optimized process of the embodiment 1; compared with the canine food control group 1 without the sterilization probiotics group, the canine food (experimental group 1 and experimental group 2) with the sterilization probiotics added and lower hardness is more beneficial to the absorption of nutrient substances, wherein the feed effect of adding four probiotics compound bacteria agents is optimal.
2) Fecal forming degree
TABLE 3 stool scoring basis
Table 4-fecal scoring of beagle puppies during the trial period
| Project | Control group 1 | Control group 2 | Experiment group 1 | Experiment group 2 |
| Day 0 | 3.61±0.27 | 3.61±0.59 | 3.62±0.21 | 3.63±0.18 |
| Day 12 | 3.53±0.94 | 3.73±0.87 | 3.59±0.38 | 3.64±0.45 |
| Day 24 | 3.60±0.53 | 3.67±0.48 | 3.69±0.42 | 3.70±0.35 |
| Day 36 | 3.66±0.43 | 3.66±0.53 | 3.88±0.31 | 3.92±0.26 |
| Day 48 | 3.70±0.35 | 3.57±0.56 | 3.91±0.23 | 4.02±0.15 |
| Day 60 | 3.74±0.34 | 3.83±0.38 | 3.99±0.19 | 4.05±0.12 |
From the experimental results, the fecal molding degree of the experimental group 1 and the experimental group 2 in the previous 20 days is not much different from that of the control group, but after 30 days, the fecal score is obviously better than that of the control group, namely, the addition of the feed for inactivating probiotics is more beneficial to fecal molding, the condition of soft feces is reduced, and the feed effect of adding four probiotics compound bactericides is optimal.
3) Diarrhea rate
TABLE 5 diarrhea Rate of beagle puppies during the trial (including soft stool)
/>
The experimental results prove that the effect of the experimental group feed is different from the feeding effect of the common feed to a certain extent, the experimental group feed can inhibit diarrhea and soft feces to a greater extent in the experimental process, and the feed effect of adding four probiotics compound microbial inoculum is optimal.
4) Intestinal flora detection: puppy faeces (experimental dogs of group 1 experimental dog food 1 and control dog food 1) were taken every month for intestinal flora detection until 4 months of age. The following table shows the results of measurement and counting of E.coli in puppy feces.
TABLE 6 E.coli content in the puppy intestinal tract (CFU/g)
| Group of | Initial detection | 3 months of age | 4 months of age |
| Experiment group 1 | 9.18±0.13 | 8.58±0.08 | 7.99±0.06 |
| Experiment group 2 | 9.16±0.12 | 8.53±0.09 | 7.94±0.07 |
| Control group 1 | 9.19±0.18 | 8.64±0.07 | 8.31±0.05 |
| Control group2 | 9.27±0.14 | 8.74±0.07 | 8.08±0.08 |
Experimental results prove that the experimental group has obvious inhibition effect on the content of the escherichia coli in the intestinal tract of the puppy, the content of the escherichia coli (particularly, the feed added with four probiotic composite bacteria agents) can be obviously reduced, and the food of the puppy without the addition of the sterilized probiotics (control group 1) has no change on the content of the escherichia coli in the intestinal tract basically, so that the addition of the sterilized probiotic bacteria agents can regulate the content of the flora in the intestinal tract and optimize the ecology of the flora in the intestinal tract.
5) Weight gain record
TABLE 7 puppy growth results (weight measurement results)
| Group of | Initial body weight | 3 months of age | 4 months of age |
| Experiment group 1 | 2.43±0.21kg | 4.18±0.15kg | 5.20±0.31kg |
| Experiment group 2 | 2.42±0.20kg | 4.23±0.16kg | 5.35±0.24kg |
| Control group 1 | 2.44±0.24kg | 4.15±0.18kg | 4.92±0.39kg |
| Control group 2 | 2.43±0.22kg | 3.91±0.14kg | 4.87±0.42kg |
During the feeding period, the puppies of the experimental group basically have no weaning stress phenomena such as diarrhea and the like, and have good weight gain. The weaning stress of the control group improved more slowly and the weight gain was relatively small. Experimental results show that the canine foods of the experimental group have good weight gain and growth and development effects, and the canine foods of the control group 1 and the control group 2 grow relatively slowly.
EXAMPLE 3 optimization of the Probiotics content of inactivated probiotics
This example optimizes the bacteria content of inactivated probiotics in a puppy diet formulation, comprising the steps of:
(1) Optimization of the bacterial content of Pediococcus acidilactici (preservation number CGMCC No. 23551)
Preliminary optimization of the bacterial content of Pediococcus acidilactici in the inactivated probiotic bacterial content: 10 11 、10 10 、10 9 、10 8 CFU/g。
The preparation method comprises the following steps of: 10 11 、10 10 、10 9 、10 8 CFU/g test dog food (sequentially named as test dog food 1-4), a diarrhea puppy system (4 in each group, male and female half) was constructed by a senna induction test, and antidiarrheal effects of dog food under different bacteria contents were tested, and the results are shown in the following Table 8:
TABLE 8 antidiarrheal effect at different levels
| Group of | Mental depression | Watery diarrhea | Appetite reduction | Weight change amount |
| Test dog food 1 | 1 | 1 | 2 | 0 |
| Test dog food 2 | 0 | 0 | 0 | +10％ |
| Test dog food 3 | 0 | 0 | 1 | +2％ |
| Test dog food 4 | 1 | 2 | 2 | -4％ |
Experiments prove that 10 10 The bacterial content of CFU/g has the best effect.
Further optimizing: the bacterial contents are respectively 1 multiplied by 10 10 、2×10 10 、4×10 10 、6×10 10 、8×10 10 CFU/g was tested. Proof 1X 10 10 -4×10 10 Has better effect between CFU/g, wherein 2×10 10 CFU/g effect is optimal.
(2) Proportion optimization of composite microbial inoculum
Determination of Pediococcus acidilactici content at 2X 10 10 CFU/g, comparing the bacteria agent of the combination of Pediococcus acidilactici/Lactobacillus reuteri/enterococcus faecalis/Lactobacillus plantarum, further optimizing the proportion of each strain in the composite bacteria agent, and the result shows that the content ratio of Pediococcus acidilactici/Lactobacillus reuteri/enterococcus faecalis/Lactobacillus plantarum is 150-200: 40-60: 2 to 4:1-2, the composite microbial inoculum has better effect, and the best combination of the effects is as follows: pediococcus acidilactici 2.0X10 10 CFU/g, lactobacillus reuteri 5.0X10 9 CFU/g, enterococcus faecalis 3.0X10 8 CFU/g, lactobacillus plantarum 2.0X10 8 CFU/g。
EXAMPLE 4 synergistic action of eucommia ulmoides leaf extract
The present example demonstrates the synergistic effect of eucommia ulmoides leaf extract and inactivated probiotics, and the test group adopted comprises:
experimental group 2 (puppy diet 2 prepared in example 2): feed without eucommia ulmoides leaf extract and inactivated probiotics (four probiotics composite microbial agents);
experimental group 3 (puppy diet 3 prepared in example 2): feed containing folium Eucommiae extract and inactivated probiotics (four probiotic compound bacteria);
control group 1 (prepared without eucommia ulmoides leaf extract and inactivated probiotics, example 2 method): the feed (15 parts of puffed corn, 3 parts of puffed soybean meal, 20.55 parts of puffed rice flour, 30 parts of chicken powder, 7 parts of olive oil, 7 parts of duck oil, 0.8 part of stone powder, 0.05 part of choline chloride, 0.3 part of sodium chloride, 2 parts of chicken liver powder, 5 parts of beef flavor, 5 parts of milk powder and 4 parts of multi-vitamin multi-mineral premix) prepared by adopting the preparation method of the embodiment 1 does not contain eucommia ulmoides leaf extract and inactivated probiotics;
control group 3 (containing eucommia ulmoides leaf extract, without inactivated probiotics, prepared by the method of example 2): the feed composition was the same as puppy diet 3, but no inactivated probiotic group agent was added.
Puppy diarrhea treatment experiments were performed using the above feed. 16 healthy and conformably weaned puppies of 4 months of age were selected into 4 groups of 4, each with senna leaf induced diarrhea. Feeding the experimental group 2 dog food, the experimental group 3 dog food, the control group 1 dog food and the control group 3 dog food respectively, and observing the condition of puppies on the 2 nd day and the 3 rd day and the weight change condition on the 5 th day.
TABLE 9 next day puppy condition
| Group of | Mental depression | Watery diarrhea | Appetite reduction |
| Experiment group 2 | 2 | 2 | 1 |
| Experiment group 3 | 0 | 0 | 0 |
| Control group 1 | 4 | 4 | 4 |
| Control group 3 | 3 | 2 | 3 |
TABLE 10 third day puppy condition and fifth day weight change
| Group of | Mental depression | Watery diarrhea | Appetite reduction | Weight change amount |
| Experiment group 2 | 0 | 0 | 0 | +4％ |
| Experiment group 3 | 0 | 0 | 0 | +6％ |
| Control group 1 | 3 | 4 | 4 | -6％ |
| Control group 3 | 2 | 1 | 2 | +1％ |
From the above results, it is clear that the addition of eucommia ulmoides leaf extract alone has a certain effect of reducing diarrhea rate, but the effect is not as good as that of the addition of inactivated probiotic alone, and is not as good as that of the combination of the inactivated probiotic and eucommia ulmoides leaf extract. The experiment group 3 (4 probiotics and the folium cortex eucommiae extract are added simultaneously) has faster and better treatment effect on the stress diarrhea, which indicates that the folium cortex eucommiae extract and the sterilization probiotic preparation can be matched for use, so that the sterilization probiotic preparation has quicker effect and synergistic effect.
The above description of the specific embodiments of the present invention has been given by way of example only, and the present invention is not limited to the above described specific embodiments. Any equivalent modifications and substitutions for the present invention will occur to those skilled in the art, and are also within the scope of the present invention. Accordingly, equivalent changes and modifications are intended to be included within the scope of the present invention without departing from the spirit and scope thereof.
Claims (10)
1. An experimental puppy diet, comprising, in parts by weight: 10-30 parts of puffed corn, 1-5 parts of puffed soybean meal, 15-30 parts of puffed rice flour, 25-40 parts of chicken meal, 3-10 parts of olive oil, 3-10 parts of duck oil, 0.5-1 part of stone powder, 0.02-0.05 part of choline chloride, 0.1-0.3 part of sodium chloride, 1-4 parts of chicken liver meal, 4-8 parts of beef flavor, 4-8 parts of milk powder and 3-4 parts of multidimensional multi-mineral premix; wherein, the experimental puppy food further comprises inactivated probiotics, and 1-6g of the inactivated probiotics are added into each kg of experimental puppy food.
2. The experimental puppy food according to claim 1, wherein said inactivated probiotic comprises pediococcus acidilactici, classified under the name Pediococcus acidilactici, with a collection number of CGMCC No.23551.
3. The experimental puppy diet of claim 2 wherein said inactivated probiotics further comprise lactobacillus reuteri, enterococcus faecalis, lactobacillus plantarum.
4. The experimental puppy food according to claim 3, wherein said pediococcus acidilactici, lactobacillus reuteri, enterococcus faecalis, lactobacillus plantarum have a content ratio of 50-200:30-80:2-5:1-3.
5. The experimental puppy diet of claim 1 further comprising a plant extract, said plant extract being an eucommia ulmoides leaf extract; wherein, 4-12g of plant extract is added into each kg of experimental puppy food.
6. The experimental puppy diet of claim 1, wherein said experimental puppy diet comprises, in parts by weight: 15 parts of puffed corn, 3 parts of puffed soybean meal, 20.55 parts of puffed rice flour, 30 parts of chicken powder, 7 parts of olive oil, 7 parts of duck oil, 0.8 part of stone powder, 0.05 part of choline chloride, 0.3 part of sodium chloride, 2 parts of chicken liver powder, 5 parts of beef flavor agent, 5 parts of milk powder and 4 parts of multi-vitamin multi-mineral premix; wherein, 8g of plant extract and 3g of inactivated probiotics are added into each kg of experimental puppy food; the content ratio of the inactivated probiotics is 200:50:3:2, pediococcus acidilactici, lactobacillus reuteri, enterococcus faecalis and Lactobacillus plantarum, wherein the plant extract is eucommia ulmoides leaf extract.
7. A method of preparing the experimental puppy diet of any one of claims 1 to 6, comprising the steps of:
s1) sequentially carrying out primary mixing, primary crushing, pre-curing and secondary crushing on corn, soybean meal and rice in predetermined parts by weight to obtain a first mixture;
s2) mixing the first mixture with chicken powder, stone powder, sodium chloride, choline chloride, multidimensional multi-mineral premix and inactivated probiotics for the second time according to the preset weight parts to obtain a second mixture, and adding the second mixture in the step if the second mixture contains plant extracts;
s3) tempering the second mixture, adding predetermined parts by weight of duck oil and olive oil in the tempering process, puffing, granulating and drying to obtain a third mixture;
and S4) spraying the third mixture by using the goat milk powder and the beef flavor agent in predetermined weight parts to obtain the experimental puppy food.
8. The preparation method according to claim 7, wherein the step S3 specifically includes: tempering the second mixture at 130 ℃ under 0.5Mpa steam, and adding mixed oil of duck oil and olive oil in the tempering process; puffing and granulating after the material temperature reaches 92 ℃, and drying at 120 ℃ for 10min and 60 ℃ for 10min after forming, wherein the diameter of a circular die hole is 7 mm.
9. Use of the experimental puppy diet of any one of claims 1 to 6 or the method of preparation of any one of claims 7 to 8 for reducing the diarrhea rate of experimental puppies.
10. The use of claim 9 wherein the experimental puppy diet is fed 2 times a day in an amount of 2.5% to 3.5% of the weight of the experimental dog.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311391117.2A CN117179177A (en) | 2023-10-25 | 2023-10-25 | Experimental puppy food and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311391117.2A CN117179177A (en) | 2023-10-25 | 2023-10-25 | Experimental puppy food and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN117179177A true CN117179177A (en) | 2023-12-08 |
Family
ID=89001952
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202311391117.2A Pending CN117179177A (en) | 2023-10-25 | 2023-10-25 | Experimental puppy food and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117179177A (en) |
-
2023
- 2023-10-25 CN CN202311391117.2A patent/CN117179177A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP3609513B1 (en) | Yeast and bacterial probiotics combinations and methods of use to improve swine production | |
| Yörük et al. | The effects of supplementation of humate and probiotic on egg production and quality parameters during the late laying period in hens | |
| Brooks et al. | Liquid feeding of pigs: potential for reducing environmental impact and for improving productivity and food safety | |
| Ghosh et al. | Supplementation of an isolated fish gut bacterium, Bacillus circulans, in formulated diets for rohu, Labeo rohita, fingerlings | |
| CN104164377B (en) | Clostridium butyricum and its application | |
| CN107996823A (en) | Composite probiotic fermented feed and its preparation method and application | |
| Eid et al. | Effect of using probiotic as growth promoters in commercial diets for monosex Nile tilapia (Oreochromis niloticus) fingerlings. | |
| CN105535387B (en) | Animal and bird intestines diarrhea ferment and preparation method thereof can be improved | |
| CN109805172A (en) | A kind of composite bacteria fermentation of Chinese herbal medicine feed and preparation method thereof | |
| CN107041483A (en) | A kind of laying cycle of laying hens feed for improving premunition and preparation method thereof | |
| CN108142641A (en) | A kind of pig starter feed of piglet with natural ferment and comprising the ferment | |
| CN102885202A (en) | Novel poultry micro-ecological preparation and preparation method thereof | |
| CN104543410B (en) | Disease-resistant pig feed additive of a kind of health care and preparation method thereof | |
| Sadati et al. | Efficacy of whole wheat grain in combination with acidified drinking water on growth performance, gizzard development, intestinal morphology, and microbial population of broiler chickens | |
| Khan et al. | Effect of Lactobacillus plantarum and Lactobacillus buchneri on composition, aerobic stability, total lactic acid bacteria and E. coli count of ensiled corn stover with or without molasses supplementation. | |
| CN106615814A (en) | Feed, preparation method thereof and application of feed to increase of feed intake of lactating sows | |
| Chimsung et al. | Fermented golden apple snails as an alternative protein source in sex-reversed red tilapia (Oreochromis niloticus x O. mossambicus) diets | |
| KR102492755B1 (en) | Method for preparing fermented total mixed ration using microbial strain complex and steam treatment | |
| KR20100027280A (en) | Fermented feeds using by-products obtained from food wastes and processing method thereof | |
| CN116831210A (en) | Composite biological feed and method for improving activity of probiotics in composite biological feed | |
| CN116584591A (en) | Low-nitrogen biological feed for laying hens producing clean eggs and preparation method thereof | |
| Lebedev et al. | Formation of element status at chickens when using enzyme, probiotic and antibiotic agents in food | |
| CN117179177A (en) | Experimental puppy food and preparation method thereof | |
| CN101773204B (en) | Preparation method of traditional Chinese medicine viable bacteria feed master batch and preparation method using master batch to prepare feed | |
| CN112704153B (en) | Immunodeficient mouse feed and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |