CN111394275A - Bacillus amyloliquefaciens and application thereof, aquatic feed and aquaculture method - Google Patents
Bacillus amyloliquefaciens and application thereof, aquatic feed and aquaculture method Download PDFInfo
- Publication number
- CN111394275A CN111394275A CN202010109464.1A CN202010109464A CN111394275A CN 111394275 A CN111394275 A CN 111394275A CN 202010109464 A CN202010109464 A CN 202010109464A CN 111394275 A CN111394275 A CN 111394275A
- Authority
- CN
- China
- Prior art keywords
- bacillus amyloliquefaciens
- fish
- group
- feed
- fishes
- 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.)
- Granted
Links
- 241000193744 Bacillus amyloliquefaciens Species 0.000 title claims abstract description 70
- 238000000034 method Methods 0.000 title claims description 17
- 238000009360 aquaculture Methods 0.000 title claims description 16
- 244000144974 aquaculture Species 0.000 title claims description 16
- 241000251468 Actinopterygii Species 0.000 claims abstract description 79
- 210000004369 blood Anatomy 0.000 claims abstract description 22
- 239000008280 blood Substances 0.000 claims abstract description 22
- 230000008021 deposition Effects 0.000 claims abstract description 17
- 230000012010 growth Effects 0.000 claims abstract description 16
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 11
- 208000035240 Disease Resistance Diseases 0.000 claims abstract description 10
- 230000001737 promoting effect Effects 0.000 claims abstract description 10
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 10
- 238000004321 preservation Methods 0.000 claims abstract description 7
- 230000003385 bacteriostatic effect Effects 0.000 claims description 7
- 238000002360 preparation method Methods 0.000 claims description 7
- 239000000203 mixture Substances 0.000 claims description 6
- 238000009472 formulation Methods 0.000 claims description 2
- 230000014101 glucose homeostasis Effects 0.000 claims 1
- 235000000346 sugar Nutrition 0.000 abstract description 16
- 150000001720 carbohydrates Chemical class 0.000 abstract description 12
- 235000014633 carbohydrates Nutrition 0.000 abstract description 12
- 230000002159 abnormal effect Effects 0.000 abstract description 6
- 230000006870 function Effects 0.000 abstract description 5
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 24
- 210000004185 liver Anatomy 0.000 description 24
- 230000000694 effects Effects 0.000 description 22
- 229920002472 Starch Polymers 0.000 description 21
- 239000008107 starch Substances 0.000 description 21
- 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 19
- 239000008103 glucose Substances 0.000 description 19
- 239000001963 growth medium Substances 0.000 description 19
- 235000019698 starch Nutrition 0.000 description 19
- 239000000243 solution Substances 0.000 description 17
- 239000006041 probiotic Substances 0.000 description 15
- 235000018291 probiotics Nutrition 0.000 description 15
- 230000000529 probiotic effect Effects 0.000 description 13
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 13
- 239000007788 liquid Substances 0.000 description 11
- 239000003925 fat Substances 0.000 description 10
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 description 10
- 239000004382 Amylase Substances 0.000 description 8
- 102000013142 Amylases Human genes 0.000 description 8
- 108010065511 Amylases Proteins 0.000 description 8
- 241000894006 Bacteria Species 0.000 description 8
- 210000000579 abdominal fat Anatomy 0.000 description 8
- 235000019418 amylase Nutrition 0.000 description 8
- 230000001580 bacterial effect Effects 0.000 description 8
- 241000607528 Aeromonas hydrophila Species 0.000 description 7
- 230000003247 decreasing effect Effects 0.000 description 7
- 239000007787 solid Substances 0.000 description 7
- 230000004584 weight gain Effects 0.000 description 7
- 235000019786 weight gain Nutrition 0.000 description 7
- 101000878595 Arabidopsis thaliana Squalene synthase 1 Proteins 0.000 description 6
- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Chemical compound CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 description 6
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 6
- 239000003833 bile salt Substances 0.000 description 6
- XBDQKXXYIPTUBI-UHFFFAOYSA-N dimethylselenoniopropionate Natural products CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 description 6
- 238000001243 protein synthesis Methods 0.000 description 6
- 210000002966 serum Anatomy 0.000 description 6
- 230000004083 survival effect Effects 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 230000014616 translation Effects 0.000 description 6
- 241000276707 Tilapia Species 0.000 description 5
- 238000002835 absorbance Methods 0.000 description 5
- 210000001789 adipocyte Anatomy 0.000 description 5
- 210000004027 cell Anatomy 0.000 description 5
- 239000012153 distilled water Substances 0.000 description 5
- 238000000338 in vitro Methods 0.000 description 5
- 210000003734 kidney Anatomy 0.000 description 5
- 238000009630 liquid culture Methods 0.000 description 5
- 210000002540 macrophage Anatomy 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 241000193830 Bacillus <bacterium> Species 0.000 description 4
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 4
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 4
- 108010065917 TOR Serine-Threonine Kinases Proteins 0.000 description 4
- 230000009286 beneficial effect Effects 0.000 description 4
- 230000008901 benefit Effects 0.000 description 4
- 239000007853 buffer solution Substances 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 238000002156 mixing Methods 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 244000153158 Ammi visnaga Species 0.000 description 3
- 235000010585 Ammi visnaga Nutrition 0.000 description 3
- 108090000565 Capsid Proteins Proteins 0.000 description 3
- 102100023321 Ceruloplasmin Human genes 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- 206010018910 Haemolysis Diseases 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 102000013530 TOR Serine-Threonine Kinases Human genes 0.000 description 3
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 229960001701 chloroform Drugs 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 238000007598 dipping method Methods 0.000 description 3
- 238000005265 energy consumption Methods 0.000 description 3
- 230000014509 gene expression Effects 0.000 description 3
- 239000011521 glass Substances 0.000 description 3
- 230000034659 glycolysis Effects 0.000 description 3
- 230000008588 hemolysis Effects 0.000 description 3
- 210000000987 immune system Anatomy 0.000 description 3
- 238000002347 injection Methods 0.000 description 3
- 239000007924 injection Substances 0.000 description 3
- 230000014759 maintenance of location Effects 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 239000001301 oxygen Substances 0.000 description 3
- 229910052760 oxygen Inorganic materials 0.000 description 3
- 230000037361 pathway Effects 0.000 description 3
- 235000019260 propionic acid Nutrition 0.000 description 3
- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- 238000005303 weighing Methods 0.000 description 3
- 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 2
- GHOKWGTUZJEAQD-ZETCQYMHSA-N (D)-(+)-Pantothenic acid Chemical compound OCC(C)(C)[C@@H](O)C(=O)NCCC(O)=O GHOKWGTUZJEAQD-ZETCQYMHSA-N 0.000 description 2
- GVJHHUAWPYXKBD-UHFFFAOYSA-N (±)-α-Tocopherol Chemical compound OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-UHFFFAOYSA-N 0.000 description 2
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- 241000283690 Bos taurus Species 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- MJVAVZPDRWSRRC-UHFFFAOYSA-N Menadione Chemical compound C1=CC=C2C(=O)C(C)=CC(=O)C2=C1 MJVAVZPDRWSRRC-UHFFFAOYSA-N 0.000 description 2
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 description 2
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 2
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 210000000683 abdominal cavity Anatomy 0.000 description 2
- 230000003187 abdominal effect Effects 0.000 description 2
- 230000003213 activating effect Effects 0.000 description 2
- 230000003321 amplification Effects 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 244000052616 bacterial pathogen Species 0.000 description 2
- 230000037396 body weight Effects 0.000 description 2
- 238000009835 boiling Methods 0.000 description 2
- WORJEOGGNQDSOE-UHFFFAOYSA-N chloroform;methanol Chemical compound OC.ClC(Cl)Cl WORJEOGGNQDSOE-UHFFFAOYSA-N 0.000 description 2
- 238000013375 chromatographic separation Methods 0.000 description 2
- 230000034994 death Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 230000006862 enzymatic digestion Effects 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 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 2
- 239000007789 gas Substances 0.000 description 2
- 238000004817 gas chromatography Methods 0.000 description 2
- 210000001035 gastrointestinal tract Anatomy 0.000 description 2
- 230000013632 homeostatic 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
- 210000005228 liver tissue Anatomy 0.000 description 2
- 230000002503 metabolic effect Effects 0.000 description 2
- 239000002207 metabolite Substances 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 238000003199 nucleic acid amplification method Methods 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 239000012074 organic phase Substances 0.000 description 2
- 230000010355 oscillation Effects 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- LXNHXLLTXMVWPM-UHFFFAOYSA-N pyridoxine Chemical compound CC1=NC=C(CO)C(CO)=C1O LXNHXLLTXMVWPM-UHFFFAOYSA-N 0.000 description 2
- 239000011541 reaction mixture Substances 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 230000019254 respiratory burst Effects 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 238000009423 ventilation Methods 0.000 description 2
- 238000003260 vortexing Methods 0.000 description 2
- FPIPGXGPPPQFEQ-UHFFFAOYSA-N 13-cis retinol Natural products OCC=C(C)C=CC=C(C)C=CC1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-UHFFFAOYSA-N 0.000 description 1
- 108020004465 16S ribosomal RNA Proteins 0.000 description 1
- LWFUFLREGJMOIZ-UHFFFAOYSA-N 3,5-dinitrosalicylic acid Chemical compound OC(=O)C1=CC([N+]([O-])=O)=CC([N+]([O-])=O)=C1O LWFUFLREGJMOIZ-UHFFFAOYSA-N 0.000 description 1
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 229920000936 Agarose Polymers 0.000 description 1
- GHOKWGTUZJEAQD-UHFFFAOYSA-N Chick antidermatitis factor Natural products OCC(C)(C)C(O)C(=O)NCCC(O)=O GHOKWGTUZJEAQD-UHFFFAOYSA-N 0.000 description 1
- AUNGANRZJHBGPY-UHFFFAOYSA-N D-Lyxoflavin Natural products OCC(O)C(O)C(O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O AUNGANRZJHBGPY-UHFFFAOYSA-N 0.000 description 1
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 1
- 108010028690 Fish Proteins Proteins 0.000 description 1
- 235000019733 Fish meal Nutrition 0.000 description 1
- 238000003794 Gram staining Methods 0.000 description 1
- SQUHHTBVTRBESD-UHFFFAOYSA-N Hexa-Ac-myo-Inositol Natural products CC(=O)OC1C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C1OC(C)=O SQUHHTBVTRBESD-UHFFFAOYSA-N 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 238000007696 Kjeldahl method Methods 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 101150097381 Mtor gene Proteins 0.000 description 1
- 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 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- AUNGANRZJHBGPY-SCRDCRAPSA-N Riboflavin Chemical compound OC[C@@H](O)[C@@H](O)[C@@H](O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O AUNGANRZJHBGPY-SCRDCRAPSA-N 0.000 description 1
- 241001052560 Thallis Species 0.000 description 1
- GLNADSQYFUSGOU-GPTZEZBUSA-J Trypan blue Chemical compound [Na+].[Na+].[Na+].[Na+].C1=C(S([O-])(=O)=O)C=C2C=C(S([O-])(=O)=O)C(/N=N/C3=CC=C(C=C3C)C=3C=C(C(=CC=3)\N=N\C=3C(=CC4=CC(=CC(N)=C4C=3O)S([O-])(=O)=O)S([O-])(=O)=O)C)=C(O)C2=C1N GLNADSQYFUSGOU-GPTZEZBUSA-J 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- FPIPGXGPPPQFEQ-BOOMUCAASA-N Vitamin A Natural products OC/C=C(/C)\C=C\C=C(\C)/C=C/C1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-BOOMUCAASA-N 0.000 description 1
- 229930003270 Vitamin B Natural products 0.000 description 1
- 229930003451 Vitamin B1 Natural products 0.000 description 1
- 229930003471 Vitamin B2 Natural products 0.000 description 1
- 229930003268 Vitamin C Natural products 0.000 description 1
- QYSXJUFSXHHAJI-XFEUOLMDSA-N Vitamin D3 Natural products C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)CCCC(C)C)=C/C=C1\C[C@@H](O)CCC1=C QYSXJUFSXHHAJI-XFEUOLMDSA-N 0.000 description 1
- 229930003427 Vitamin E Natural products 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 210000000577 adipose tissue Anatomy 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- FPIPGXGPPPQFEQ-OVSJKPMPSA-N all-trans-retinol Chemical compound OC\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-OVSJKPMPSA-N 0.000 description 1
- 239000011609 ammonium molybdate Substances 0.000 description 1
- 235000018660 ammonium molybdate Nutrition 0.000 description 1
- APUPEJJSWDHEBO-UHFFFAOYSA-P ammonium molybdate Chemical compound [NH4+].[NH4+].[O-][Mo]([O-])(=O)=O APUPEJJSWDHEBO-UHFFFAOYSA-P 0.000 description 1
- 229940010552 ammonium molybdate Drugs 0.000 description 1
- 239000003674 animal food additive Substances 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 229960002685 biotin Drugs 0.000 description 1
- 235000020958 biotin Nutrition 0.000 description 1
- 239000011616 biotin Substances 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 210000000234 capsid Anatomy 0.000 description 1
- 229960001231 choline Drugs 0.000 description 1
- OEYIOHPDSNJKLS-UHFFFAOYSA-N choline Chemical compound C[N+](C)(C)CCO OEYIOHPDSNJKLS-UHFFFAOYSA-N 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- GFHNAMRJFCEERV-UHFFFAOYSA-L cobalt chloride hexahydrate Chemical compound O.O.O.O.O.O.[Cl-].[Cl-].[Co+2] GFHNAMRJFCEERV-UHFFFAOYSA-L 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 229910000365 copper sulfate Inorganic materials 0.000 description 1
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 102000038379 digestive enzymes Human genes 0.000 description 1
- 108091007734 digestive enzymes Proteins 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- BVTBRVFYZUCAKH-UHFFFAOYSA-L disodium selenite Chemical compound [Na+].[Na+].[O-][Se]([O-])=O BVTBRVFYZUCAKH-UHFFFAOYSA-L 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000004222 ferrous gluconate Substances 0.000 description 1
- 235000013924 ferrous gluconate Nutrition 0.000 description 1
- 229960001645 ferrous gluconate Drugs 0.000 description 1
- 235000021323 fish oil Nutrition 0.000 description 1
- 239000004467 fishmeal Substances 0.000 description 1
- 239000000834 fixative Substances 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 229960000304 folic acid Drugs 0.000 description 1
- 235000019152 folic acid Nutrition 0.000 description 1
- 239000011724 folic acid Substances 0.000 description 1
- WIGCFUFOHFEKBI-UHFFFAOYSA-N gamma-tocopherol Natural products CC(C)CCCC(C)CCCC(C)CCCC1CCC2C(C)C(O)C(C)C(C)C2O1 WIGCFUFOHFEKBI-UHFFFAOYSA-N 0.000 description 1
- 210000003736 gastrointestinal content Anatomy 0.000 description 1
- 230000009036 growth inhibition Effects 0.000 description 1
- 230000007366 host health Effects 0.000 description 1
- 235000003642 hunger Nutrition 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 210000002865 immune cell Anatomy 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 229960000367 inositol Drugs 0.000 description 1
- CDAISMWEOUEBRE-GPIVLXJGSA-N inositol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](O)[C@@H]1O CDAISMWEOUEBRE-GPIVLXJGSA-N 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 210000004347 intestinal mucosa Anatomy 0.000 description 1
- 230000006799 invasive growth in response to glucose limitation Effects 0.000 description 1
- 239000011630 iodine Substances 0.000 description 1
- 229910052740 iodine Inorganic materials 0.000 description 1
- VRIVJOXICYMTAG-IYEMJOQQSA-L iron(ii) gluconate Chemical compound [Fe+2].OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C([O-])=O.OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C([O-])=O VRIVJOXICYMTAG-IYEMJOQQSA-L 0.000 description 1
- 230000001788 irregular Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 description 1
- 229940061634 magnesium sulfate heptahydrate Drugs 0.000 description 1
- ISPYRSDWRDQNSW-UHFFFAOYSA-L manganese(II) sulfate monohydrate Chemical compound O.[Mn+2].[O-]S([O-])(=O)=O ISPYRSDWRDQNSW-UHFFFAOYSA-L 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 208000030159 metabolic disease Diseases 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 229960003512 nicotinic acid Drugs 0.000 description 1
- 235000001968 nicotinic acid Nutrition 0.000 description 1
- 239000011664 nicotinic acid Substances 0.000 description 1
- 208000019180 nutritional disease Diseases 0.000 description 1
- 230000020477 pH reduction Effects 0.000 description 1
- 229940055726 pantothenic acid Drugs 0.000 description 1
- 235000019161 pantothenic acid Nutrition 0.000 description 1
- 239000011713 pantothenic acid Substances 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 230000007918 pathogenicity Effects 0.000 description 1
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 239000001103 potassium chloride Substances 0.000 description 1
- 235000011164 potassium chloride Nutrition 0.000 description 1
- LJCNRYVRMXRIQR-OLXYHTOASA-L potassium sodium L-tartrate Chemical compound [Na+].[K+].[O-]C(=O)[C@H](O)[C@@H](O)C([O-])=O LJCNRYVRMXRIQR-OLXYHTOASA-L 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- RADKZDMFGJYCBB-UHFFFAOYSA-N pyridoxal hydrochloride Natural products CC1=NC=C(CO)C(C=O)=C1O RADKZDMFGJYCBB-UHFFFAOYSA-N 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 230000029058 respiratory gaseous exchange Effects 0.000 description 1
- 229960002477 riboflavin Drugs 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 description 1
- 235000021391 short chain fatty acids Nutrition 0.000 description 1
- 150000004666 short chain fatty acids Chemical class 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000001476 sodium potassium tartrate Substances 0.000 description 1
- 235000011006 sodium potassium tartrate Nutrition 0.000 description 1
- 239000011781 sodium selenite Substances 0.000 description 1
- 235000015921 sodium selenite Nutrition 0.000 description 1
- 229960001471 sodium selenite Drugs 0.000 description 1
- 230000007480 spreading Effects 0.000 description 1
- 238000003892 spreading Methods 0.000 description 1
- -1 starch is lower Chemical class 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 229960003495 thiamine Drugs 0.000 description 1
- DPJRMOMPQZCRJU-UHFFFAOYSA-M thiamine hydrochloride Chemical compound Cl.[Cl-].CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N DPJRMOMPQZCRJU-UHFFFAOYSA-M 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 239000012137 tryptone Substances 0.000 description 1
- 238000001291 vacuum drying Methods 0.000 description 1
- 210000001835 viscera Anatomy 0.000 description 1
- 235000019155 vitamin A Nutrition 0.000 description 1
- 239000011719 vitamin A Substances 0.000 description 1
- 235000019156 vitamin B Nutrition 0.000 description 1
- 239000011720 vitamin B Substances 0.000 description 1
- 235000010374 vitamin B1 Nutrition 0.000 description 1
- 239000011691 vitamin B1 Substances 0.000 description 1
- 235000019164 vitamin B2 Nutrition 0.000 description 1
- 239000011716 vitamin B2 Substances 0.000 description 1
- 235000019158 vitamin B6 Nutrition 0.000 description 1
- 239000011726 vitamin B6 Substances 0.000 description 1
- 235000019154 vitamin C Nutrition 0.000 description 1
- 239000011718 vitamin C Substances 0.000 description 1
- QYSXJUFSXHHAJI-YRZJJWOYSA-N vitamin D3 Chemical compound C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)CCCC(C)C)=C\C=C1\C[C@@H](O)CCC1=C QYSXJUFSXHHAJI-YRZJJWOYSA-N 0.000 description 1
- 235000005282 vitamin D3 Nutrition 0.000 description 1
- 239000011647 vitamin D3 Substances 0.000 description 1
- 235000019165 vitamin E Nutrition 0.000 description 1
- 239000011709 vitamin E Substances 0.000 description 1
- 229940046009 vitamin E Drugs 0.000 description 1
- 235000012711 vitamin K3 Nutrition 0.000 description 1
- 239000011652 vitamin K3 Substances 0.000 description 1
- 229940045997 vitamin a Drugs 0.000 description 1
- 229940011671 vitamin b6 Drugs 0.000 description 1
- 229940021056 vitamin d3 Drugs 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
- RZLVQBNCHSJZPX-UHFFFAOYSA-L zinc sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Zn+2].[O-]S([O-])(=O)=O RZLVQBNCHSJZPX-UHFFFAOYSA-L 0.000 description 1
Images
Classifications
-
- 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
- C12N1/205—Bacterial isolates
-
- 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
- C12R2001/07—Bacillus
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
- A01K61/10—Culture of aquatic animals of fish
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/742—Spore-forming bacteria, e.g. Bacillus coagulans, Bacillus subtilis, clostridium or Lactobacillus sporogenes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
-
- 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
-
- 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
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Microbiology (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Medicinal Chemistry (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Polymers & Plastics (AREA)
- Biochemistry (AREA)
- Biomedical Technology (AREA)
- Public Health (AREA)
- General Engineering & Computer Science (AREA)
- Animal Husbandry (AREA)
- Virology (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Oncology (AREA)
- Communicable Diseases (AREA)
- Mycology (AREA)
- Environmental Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Food Science & Technology (AREA)
- Molecular Biology (AREA)
- Marine Sciences & Fisheries (AREA)
- General Chemical & Material Sciences (AREA)
- Insects & Arthropods (AREA)
- Physiology (AREA)
- Birds (AREA)
- Biodiversity & Conservation Biology (AREA)
- Epidemiology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a Bacillus amyloliquefaciens strain, which is characterized in that the Bacillus amyloliquefaciens strain is Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) SS1 and is preserved in China center for type culture collection with the preservation number as follows: CCTCC M2020024, and the preservation date is 2020, 2 and 18. The bacillus amyloliquefaciens is high in safety, and has the functions of improving the utilization rate of carbohydrates of fishes, promoting the growth of the fishes, improving the blood sugar steady state of the fishes, relieving abnormal deposition of fat of the fishes, increasing the deposition of body protein, improving the disease resistance and the like.
Description
Technical Field
The invention relates to the field of aquaculture, in particular to bacillus amyloliquefaciens and application thereof, an aquaculture feed and an aquaculture method.
Background
With the continuous expansion of aquaculture scale and the annual increase of yield in China, fish meal and fish oil resources are increasingly in short supply, so that the improvement of the utilization efficiency of the existing feed raw materials is an important direction for relieving the shortage of the feed raw materials. Carbohydrates are one of the important metabolic energy supply substrates for fish and are also a cheaper energy source in aquatic feeds. Researches show that the addition of a proper amount of starch in the feed is not only beneficial to the forming of feed particles, but also beneficial to saving feed protein, reducing feed cost and reducing nitrogen emission of fish bodies. However, compared with higher animals, the digestion and utilization efficiency of the fish to the carbohydrate such as starch is lower, and the nutritional diseases such as the growth inhibition, the low feed utilization efficiency, the abnormal fat deposition, even the high death rate and the like of the fish body can be caused by taking the feed with high carbohydrate level for a long time. Therefore, the utilization efficiency of the carbohydrates by the fishes is improved, the growth of the fishes can be promoted, the feed protein is saved, the culture cost is reduced, and the economic benefit is improved.
In recent years, many studies have shown that fish gut microorganisms play an important role in promoting host health. The intestinal microorganisms not only affect the digestion of feed, the absorption of nutrient substances and the energy supply, but also regulate and control the normal physiological functions of a host and the occurrence and development of diseases. Therefore, as a substitute for antibiotics, probiotics are widely applied to the fields of livestock breeding, aquaculture, medicine and the like, can inhibit the growth of harmful bacteria, promote the development of immune system, produce beneficial metabolites, provide nutrients and energy for organisms, regulate metabolic disorders, protect intestinal mucosa barriers and the like.
At present, in the aquaculture process, probiotics are added into feed or water, so that researches for adjusting the metabolic stability of organisms and improving disease resistance are increasing, wherein bacillus and lactic acid bacteria are used more frequently. The bacillus is one of the most abundant and important strains of active products in a microorganism bank, has short growth period and very high propagation speed, and produces various metabolites in the growth and propagation processes.
However, the efficacy of the bacillus for aquatic products is mainly focused on improving disease resistance, and the effect of the bacillus for aquatic products on the regulation of fish metabolism, particularly on the aspect of promoting the utilization of carbohydrates in fish bodies, is rarely reported.
Disclosure of Invention
The bacillus amyloliquefaciens is high in safety and has the functions of improving the utilization rate of carbohydrates of fishes, promoting the growth of the fishes, improving the blood sugar steady state of the fishes, relieving abnormal deposition of fats of the fishes, increasing the deposition of body proteins, improving the disease resistance and the like.
In order to achieve the purpose, the invention adopts the following technical scheme:
provides a bacillus amyloliquefaciens strain, wherein the bacillus amyloliquefaciens strain is bacillus amyloliquefaciens (Bacillus amyloliquefaciens) SS1 and is preserved in China center for type culture collection with the preservation number as follows: CCTCC M2020024, and the preservation date is 1 month and 7 days in 2020.
Provides an application of the bacillus amyloliquefaciens SS1 in preparing a bacteriostatic preparation.
Provides an application of the bacillus amyloliquefaciens SS1 in promoting fish growth.
Provides an application of the bacillus amyloliquefaciens SS1 in improving the blood sugar homeostasis of fishes.
Provides an application of the bacillus amyloliquefaciens SS1 in reducing the fat deposition of fish.
Provides an application of the bacillus amyloliquefaciens SS1 in promoting the protein deposition of fish bodies.
Provides an application of the bacillus amyloliquefaciens SS1 in improving the disease resistance of fish.
An aquatic feed product is provided, which comprises the bacillus amyloliquefaciens SS 1.
Preferably, the concentration of the bacillus amyloliquefaciens SS1 in the aquatic feed is 105-107CFU/g。
Provides an aquaculture method, which adopts the aquatic feed to feed fishes 1-2 times per day, wherein the aquaculture time is 5-10 weeks.
The invention has at least the following beneficial effects:
the bacillus amyloliquefaciens SS1 can be added into fish culture feed to improve the utilization efficiency of carbohydrates by fish and improve the digestive enzyme activity to promote growth by the bacillus amyloliquefaciens SS 1; further, the Bacillus amyloliquefaciens SS1 can increase glycolysis level of liver of fish, thereby improving blood sugar steady state, relieving abnormal deposition of fat of fish by increasing energy consumption, and increasing body protein deposition by activating protein synthesis pathway; in addition, the bacillus amyloliquefaciens SS1 also has bacteriostatic property and can improve the disease resistance of fish bodies by protecting the immune system of organisms.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the description of the embodiments are briefly introduced below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings based on these drawings without creative efforts.
FIG. 1 shows the colony morphology of Bacillus amyloliquefaciens SS1 according to the present invention;
FIG. 2 shows the growth of Bacillus amyloliquefaciens SS1 according to the present invention on starch medium, the hemolysis, the Aeromonas hydrophila-inhibiting, and the growth on L B solid medium containing 0.3% bovine bile salt;
FIG. 3 is data of the amylase activity produced by Bacillus amyloliquefaciens SS1 of the present invention;
FIG. 4a shows the results of gas chromatographic separation of acetic acid, propionic acid and butyric acid;
FIG. 4b shows the result of gas chromatographic separation of acetic acid produced in vitro by Bacillus amyloliquefaciens SS1 of the present invention;
FIG. 5 shows the effect of Bacillus amyloliquefaciens SS1 of the present invention on the average fish body weight (a), weight gain (b), and feed efficiency (c);
FIG. 6 is a graph showing the effect of Bacillus amyloliquefaciens SS1 of the present invention on fasting blood glucose levels (a), sugar tolerance (b), and AUC (c) in fish;
FIG. 7 shows the effect of Bacillus amyloliquefaciens SS1 of the present invention on fish abdominal fat factor (MFI) (a), serum Triglyceride (TG) (b), abdominal fat cell size (c) and abdominal fat cell size assessment (d);
FIG. 8 is a graph showing the effect of Bacillus amyloliquefaciens SS1 of the present invention on fish liver fat content (a), liver TG (b), liver adipocyte size (c), and liver fat area assessment (d);
FIG. 9 shows the effect of Bacillus amyloliquefaciens SS1 on fish body hull ratio (a), hull protein (b), and mTOR gene expression level (c);
FIG. 10 is a graph showing the effect of Bacillus amyloliquefaciens SS1 of the present invention on fish survival rate (a) and head kidney macrophage oxygen respiratory burst activity (b).
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is described in further detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
Example 1:
the embodiment provides a strain of Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) SS1 (hereinafter referred to as 'SS 1'), which is preserved in China center for type culture Collection and has the preservation number: CCTCC M2020024, the preservation date is: year 2020, 1, 7.
The information of the above strains is as follows:
1. the source of the strain
The strain is isolated from intestinal tract of healthy tilapia (Oreochromys mossambicaus).
2. Morphological characteristics
Gram staining was positive, as shown in FIG. 1, the colonies after 12h of inverted culture on L B medium plates at 28 ℃ were irregular white circles with dry surfaces and volcano-like white bumps in the center of the colonies, similar to a fried egg.
3. Enzyme producing Properties
1) A single colony of SS1 was dipped with a toothpick and spotted on a starch culture medium, after 12 hours of inverted culture at 28 ℃, Lugol iodine solution was dropped on the culture plate, and the plate was left to stand for 3 to 5 minutes, and it was observed that SS1 was able to produce a transparent circle on the starch culture medium, and the result is shown in FIG. 2 ①, thus demonstrating that SS1 can produce amylase in vitro.
2) Method for measuring amylase activity by DNS reducing sugar method
A standard glucose solution (2mg/m L) is prepared by accurately weighing 0.2g glucose, dissolving in distilled water, accurately metering to 100m L with volumetric flask, mixing, and storing in refrigerator at 4 deg.C.
3, 5-dinitrosalicylic acid (DNS) reagent preparation 6.3g of DNS and 262m L2 mol/L NaOH solution are added to a 500m L hot aqueous solution containing 185g of sodium potassium tartrate, 5g of crystalline phenol and 5g of Na are added2SO3Stirring to dissolve, cooling, adding distilled water to a constant volume of 1L, and storing in a brown bottle for one week in dark place.
The specific operation method comprises the following steps:
6 clean graduated test tubes are respectively numbered, glucose solutions with the concentrations of 0mg/m L, 0.4mg/m L0, 0.8mg/m L, 1.2mg/m L, 1.6mg/m L and 1mg/m L are respectively prepared by using a standard glucose solution (2mg/m L), the glucose solutions with different concentrations of 1m L are respectively mixed with 1.5m L DNS reagent, the mixture is immediately placed on ice after being heated in boiling water bath for 5min, finally distilled water is used for fixing the volume to 10m L, the absorbance value of the reaction mixture is measured at OD 540nm after the mixture is vertically reversed and mixed, and finally, the absorbance value (540 OD) is taken as an abscissa and the glucose concentration (mg/m L) is taken as an ordinate for drawing to draw a glucose standard curve.
Taking 1m L crude enzyme solution (namely SS1 bacterial solution) and 1m L culture medium (used as a contrast) to be respectively and uniformly mixed with a substrate solution (1% soluble starch solution) and then react in a water bath at 45 ℃ for 30min, establishing three parallels, immediately absorbing 1m L reaction solution and mixing with 1.5m L DNS reagent after the reaction is finished, immediately placing on ice after boiling water bath is 5min, finally, using distilled water to fix the volume to 10m L, and measuring the absorbance value of the reaction mixture at OD 540nm after the mixture is vertically inverted and uniformly mixed.
The glucose standard curve is: y is 0.1227x-0.0073 (R)2＝0.9938)
Amylase activity (U/m L) ═ k × OD × 1000 × n/(180 × 30)
Wherein k is the slope of the glucose standard curve; OD is absorbance at 540 nm; 1000 is a unit conversion multiple; n is the dilution multiple; 180 is the molecular weight of glucose; and 30 is reaction time (min).
As shown in FIG. 3, the amylase produced in vitro from SS1 had a high amylase activity (approximately 1.0U/L).
4. Characteristic of acetic acid production
The standard is prepared by diluting acetic acid, propionic acid, and butyric acid with distilled water to 1000 times of mixed standard acid, adding 100 μ L50% sulfuric acid, acidifying, vortexing for 30s, adding 400 μ L diethyl ether, vortexing for 15s, standing for 2min, extracting, centrifuging at 4 deg.C and 12000rpm/min for 5min, and collecting the upper organic phase into 1m L brown vial.
Sample preparation, using 200 mu L SS1 bacterial liquid or corresponding culture medium as a control, adding 100 mu L50% sulfuric acid for acidification, carrying out vortex oscillation for 30s, adding 400 mu L diethyl ether, carrying out vortex oscillation for 15s, standing for 2min for extraction, centrifuging for 5min at 4 ℃ and 12000rpm/min, and absorbing an upper organic phase into a 1m L brown vial.
The gas chromatography conditions comprise initial temperature of the chromatographic column of 100 deg.C, holding for 2min, 5 deg.C raising to 180 deg.C per minute, holding for 2min, controlling flow rate of the chromatographic column to be 1m L/min, injection port temperature of 220 deg.C, no shunt, injection of 1 μ L, detector temperature of 200 deg.C, air of 200m L/min, hydrogen of 32m L/min, and nitrogen of 24m L/min.
As shown in FIG. 4a, 3 short chain fatty acids can be effectively separated by gas chromatography column, wherein the acetic acid retention time is 7.019min, the propionic acid retention time is 8.622min, and the butyric acid retention time is 10.422 min; in addition, as shown in fig. 4b, SS1 shows a relatively distinct peak at 7.020min, which indicates that SS1 can ferment starch in vitro to produce acetic acid, and the acetic acid can provide energy for the body and has an important role in reducing blood sugar level and fat deposition.
5. Pathogenicity
Inoculating the frozen strain SS1 and the separated Aeromonas hydrophila CS4(A.hyd CS4) into L B liquid culture medium according to the ratio of 1:1000 respectively for resuscitation, shaking the strain for 12h under the conditions of 28 ℃ and 220rpm/min, dipping the Aeromonas hydrophila strain liquid by using an inoculating loop, streaking on a L B culture medium plate, carrying out inverted culture at 28 ℃ for 12h, dipping an SS1 single colony by using a toothpick, spotting on a blood plate, and carrying out inverted culture at 28 ℃ for 12 h.
As shown in fig. 2 ②, SS1 (i.e., "1" in fig. 2 ②) did not cause hemolysis on the blood plate, while pathogenic bacteria a.hydcs4 (i.e., "2" and "3" in fig. 2 ②) produced a hemolysis loop on the blood plate, thereby indicating that SS1 is not significantly pathogenic and has high safety.
6. In vitro bacteriostatic properties
The SS1 bacterial liquid was adjusted to a concentration of 1 × 10 with PBS buffer6CFU/m L, centrifuging the Aeromonas hydrophila solution at 12000rpm/min, collecting precipitate, resuspending with PBS buffer solution, and adjusting concentration to 1 × 109CFU/m L, spreading 100 μ L on L B culture medium, drying the surface of the culture medium, perforating the culture medium with a perforator, picking out excess agar with a sterile needle, adding SS1 bacterial solution 20 μ L into the holes, and culturing in 28 deg.C incubator for 12 h.
From FIG. 2 ③, it can be observed that SS1 can inhibit the growth of Aeromonas hydrophila, and produce a bacteriostatic transparent zone, which indicates that the bacterium has a certain bacteriostatic action.
7. Stress resistance characteristic
Preparing L B solid culture medium containing 0.3% of ox bile salt, dipping a single colony of SS1 on L B solid culture medium with a sterile toothpick, spotting on L B solid culture medium containing 0.3% of ox bile salt, and performing inverted culture at 28 ℃ for 12 h.
From FIG. 2 ④, it can be observed that SS1 can grow on L B solid medium containing 0.3% of bovine bile salt, and produce transparent circles resistant to bile salt, indicating that the bacterium has better bile salt resistance.
Example 2:
this example provides a method of isolation of Bacillus amyloliquefaciens SS1 (hereinafter "SS 1") described in example 1, comprising the steps of:
s1, tryptone 5 g/L, Yeast extract 2 g/L2PO42g/L，MgSO4·7H2Preparing a starch culture medium by using O2 g/L5 g/L, soluble starch 10 g/L and agarose 20 g/L, and pouring the mixture in a super clean bench after autoclaving to obtain a starch culture medium solid plate;
s2, mixing intestinal contents of fish (such as tilapia) with PBS buffer solution uniformly according to a ratio of 1: 9(w/v, g/ml), centrifuging at 1500rpm/min, and coating supernatant fluid of 100 mu L on the starch culture medium solid plate;
s3, carrying out inverted culture at 28 ℃ for 24h, then screening colonies according to the size, shape and color of the colonies, preserving and numbering the inclined planes of the screened colonies, and carrying out 16S rRNA molecular identification on the screened strains to obtain the bacillus amyloliquefaciens SS 1.
Example 3:
this example provides a method for aquaculture using bacillus amyloliquefaciens SS1 (hereinafter referred to as "SS 1") described in example 1, which specifically includes the following steps:
s1, temporarily culturing the fry: purchasing 600 tails of fish fries (such as tilapia fries), temporarily culturing for 2 weeks by using commercial feed, then putting into aerated water at 28 ℃ for temporary culture, changing water every two days, and ending the temporary culture after the fish fries adapt to a growth environment and no diseases are determined; the fry is divided into five groups, namely a normal group (CON), a high starch group (HCD), a probiotic group 1(HCB1), a probiotic group 2(HCB3) and a probiotic group 3(HCB3), wherein each group comprises three parallel groups, each group comprises 30 parallel tails, and the weight of each group is 1.6 +/-0.1 g;
s2 and SS1 bacterial liquid preparation, namely inoculating the bacillus amyloliquefaciens SS1 into L B liquid culture medium according to the ratio of 1:1000(w/v, g/ml) for resuscitation, shaking the bacteria for 12 hours at 28 ℃ and 220rpm/min to obtain resuscitative bacterial liquid, inoculating the resuscitative bacterial liquid into L B liquid culture medium according to the ratio of 1:100(v/v, ml/ml) for amplification culture, shaking the bacteria for 24 hours at 28 ℃ and 220rpm/min, centrifuging for 20 minutes at 4 ℃ and 12000rpm/min, collecting thalli, and carrying out heavy suspension by PBS buffer solution to obtain heavy-suspension bacterial liquid;
s3, preparing feed: uniformly mixing the heavy-suspension bacterium liquid and high-starch feed (containing 45% of starch in a weight ratio) in a ratio of 1:2(v/w, ml/g) to ensure that the concentrations of the bacillus amyloliquefaciens SS1 in the feed are respectively 105CFU/g、106CFU/g、107CFU/g to obtain probiotic feed (namely aquatic feed) with three different strain concentrations, and then granulating the probiotic feed;
s4, feeding: the normal group (CON), the high starch group (HCD) and the probiotic group 1(HCB1) (feeding concentration 10) were fed with commercial feed, high starch feed (containing 45% starch by weight) and three probiotic feeds with different strain concentrations5CFU/g probiotic feed), probiotic group 2(HCB3) (fed at a concentration of 106CFU/g probiotic feed), probiotic group 3(HCB3) (fed at a concentration of 107CFU/g probiotic feed) fry, fed 1-2 times per day, and fed 4% of the body weight each time; starving for 24h after 5-10 weeks (preferably 8 weeks) of culture, and weighing, collecting blood and collecting tissue samples of 10 fish in each group.
The commercial feed, the high starch feed (containing 45% by weight of starch) and the probiotic feed raw material formula were tested and shown in table 1.
TABLE 1 feed ingredient formulation Table (g)
Wherein: 1. complex (mg or IU/kg): vitamin A: 500,000i.u. (international units); vitamin D3: 50,000 i.u.; vitamin E: 2500 mg; vitamin K3: 1000 mg; vitamin B1: 5000 mg; vitamin B2: 5000 mg; vitamin B6: 5000 mg; 125000 μ g of vitamin B; inositol: 25,000 mg; folic acid: 1000 mg; pantothenic acid: 10,000 mg; biotin: 250 mg; choline: 100,000 mg; nicotinic acid: 25,000 mg; vitamin C: 10,000 mg.
2. Remineralization (g/kg): calcium carbonate: 314.0 g; potassium dihydrogen phosphate: 469.3 g; magnesium sulfate heptahydrate: 147.4 g; sodium chloride: 49.8 g; ferrous gluconate: 10.9 g; 3.12g of manganese sulfate monohydrate; ammonium molybdate: 0.06 g; zinc sulfate heptahydrate: 4.67 g; anhydrous copper sulfate: 0.62 g; cobalt chloride hexahydrate: 0.08 g; sodium selenite: 30.02 g.
The function of the bacillus amyloliquefaciens SS1 is as follows:
(1) SS1 function of promoting fish growth (including weight gain)
After the culture is finished, the total weight of each group of fish bodies is respectively weighed, and the average weight, the weight gain rate and the bait coefficient of each group are calculated.
Wherein, the average weight (g/tail) is total weight (g)/mantissa, and the weight gain ratio (%) (end weight-initial weight) is × 100/initial weight.
As shown in fig. 5(a), compared with the CON group, the average weight of the HCD group was significantly increased, and the average weight of the HCB group was also significantly increased, compared with the HCD group, and as shown in fig. 5(b), the weight gain rate of the CON group was not significantly different from that of the HCD group, but the weight gain rate of the HCB group was significantly increased, compared with that of the HCD group, which indicates that the bacillus amyloliquefaciens SS1 of the present invention can significantly and rapidly promote the weight gain of fish bodies. Further, as shown in FIG. 5(c), the bait ratio of the HCB group was decreased compared to the CON group and the HCD group, and was particularly decreased significantly compared to the HCD group,
as described above, Bacillus amyloliquefaciens SS1 has high amylase activity and can improve the utilization rate of carbohydrates, so that the digestive enzyme activity, such as the activity of amylase, can be improved by applying the Bacillus amyloliquefaciens SS1 in aquatic feeds, the growth of fish bodies can be promoted, the cost can be reduced, and the economic benefit can be increased.
Note: in fig. 5 "+" represents the difference between groups, and: p < 0.05, x: p < 0.01, x: p is less than 0.001, and the HCB group data in (a) - (c) are the average values of three groups of data, namely HCB1, HCB2 and HCB 3.
(2) Function of bacillus amyloliquefaciens SS1 on improving blood sugar homeostasis of fish bodies
1) Blood was collected, serum was collected, and fasting blood glucose concentration was measured with a glucose kit.
2) Sugar tolerance test
Each group of 5 fishes was sampled to prepare a glucose solution of 500mg/m L, glucose concentration was injected intraperitoneally at 200mg/kg per tail, blood was collected at 0h, 0.5h, 1.5h and 3h, serum was collected, blood glucose concentration was measured with a glucose kit and area under the curve (AUC) was calculated, and the results are shown in FIG. 6.
As can be seen from fig. 6(a), fasting blood glucose levels were significantly increased in the HCD group compared to the CON group, but were significantly decreased in the HCB group compared to the HCD group, indicating that blood glucose levels in fish bodies could be significantly decreased by the addition of bacillus amyloliquefaciens SS1, while the results of the sugar tolerance experiment in fig. 6(b) and AUC in fig. 6(c) demonstrate that fish bodies in the HCB group had stronger sugar tolerance.
Therefore, the glycolysis level of the liver of the fish can be increased by using the bacillus amyloliquefaciens SS1, and the blood sugar level in the fish can be reduced by producing acetic acid, so that when the fish ingests the feed with high carbohydrate level for a long time, the feed helps to stabilize the blood sugar level in the fish, improve the sugar tolerance and improve the blood sugar steady state of the fish.
Note: in fig. 6 "+" represents the difference between groups, and: p < 0.05, x: p < 0.01, x: p is less than 0.001; "#" in fig. 6(b) represents the difference between the HCB group and the HCD group, and #: p < 0.05, # #: p < 0.01, ###' #: p is less than 0.001, and the HCB group data in (a) - (c) are the average values of three groups of data, namely HCB1, HCB2 and HCB 3.
(3) Bacillus amyloliquefaciens SS1 for reducing fat deposition in liver and abdominal cavity of fish
1) Placing adipose tissues with the same parts and weight of different groups of fishes in a Bonn's stationary liquid for preparing HE slices, collecting abdominal adipose tissues and weighing; liver tissues of the same part and weight were placed in Bonn's fixative for HE slice preparation, and liver tissues were collected and weighed.
2) HE slice
A conventional paraffin section preparation method and HE dyeing.
3) Liver fat content assay and TG assay
Measuring the fat content of the liver by a chloroform-methanol method, namely measuring the fat content of the liver by 3 treatment groups, wherein each group comprises 6 parallel groups, two groups of 36 clean glass test tubes with scales of 10ml are needed, the groups are marked and weighed and peeled, about 0.5g of liver sample is added into each tube, 6ml of chloroform-methanol (chloroform: methanol is 2: 1) is added into a ventilation kitchen, a tube cover is screwed down, the tubes are kept stand overnight at 4 ℃, the sample is taken out and vibrated for 30s on a vortex oscillator, the tubes are placed into a refrigerator at 4 ℃ for standing for 1h, then 0.37 mol/L mol potassium chloride is added into the ventilation kitchen, the tubes are centrifuged at 1500rpm/min for 10min in a low-speed centrifuge after being vibrated again, a bottom layer transparent liquid is sucked out by a pasteur suction tube, the tubes are transferred into a new weighed and marked glass tube after being filtered by a funnel, filter paper is washed by trichloromethane, the test tubes are placed into a vacuum drying oven, after the trichloromethane is volatilized, the glass tubes are taken out, the tube weight is weighed, the tube weight is obtained by subtracting the tube weight recorded before the fat content of the.
Serum and liver TG detection was performed according to Nanjing kit instructions.
As can be seen from fig. 7, the CON group did not have a significant difference in celiac adipose index (MFI) from the HCD group, but the HCB group had a significant decrease in MFI compared to the HCD group; and the serum TG in the HCD group is obviously increased compared with the CON group, but the serum TG in the HCB group is obviously reduced compared with the HCD group; the results of the HE sections of abdominal adipose tissues of fish and the evaluation of the size of abdominal adipocytes (Average area value of adipocyte) demonstrated that the size of abdominal adipocytes was significantly increased in the HCD group compared to the CON group, but significantly decreased in the HCB group compared to the HCD group.
Further, as can be seen from fig. 8, the liver fat content and the liver TG content of the HCD group were significantly increased compared to the CON group, but the liver fat content and the liver TG content of the HCB group were significantly decreased compared to the HCD group, and further, the HE section of the liver of fish and the evaluation of liver fat area (L ipid simple area of liver) showed that the size of the fat cells and the liver fat area of the HCB group were significantly decreased compared to the HCD group.
Note: in fig. 7-8 "", the differences between groups are represented, and: p < 0.05, x: p < 0.01, x: p is less than 0.001, and the HCB group data in (a), (b) and (d) is the average value of three groups of HCB1, HCB2 and HCB 3; the HE slice of HCB group in FIG. 7(c) was HCB3 group (SS1 concentration: 10)7CFU/g) abdominal adipose tissue sections of fish, HE sections of HCB group in FIG. 8(c) were HCB3 group (SS1 concentration of 10)7CFU/g) liver sections of tilapia.
Therefore, the addition of the bacillus amyloliquefaciens SS1 can remarkably reduce the TG content of the liver and abdominal fat of the fish by promoting the utilization capacity of carbohydrates (such as starch) of fish bodies and increasing energy consumption so as to avoid abnormal deposition of the liver and abdominal fat of the fish.
(4) Effect of Bacillus amyloliquefaciens SS1 on fish protein deposition
The head, tail and internal organs of each group of fish bodies were removed, weighed and recorded, the capsid ratio and capsid protein content were calculated, the protein content was determined by kjeldahl method and the expression level of protein synthesis marker gene mTOR was detected by real-time fluorescence quantification method, the results are shown in fig. 9.
The results show that the body-shell ratio of the CON group and the HCD group is not obviously different, but the body-shell ratio of the HCB group is obviously increased, which indicates that the protein content is obviously increased; meanwhile, the results of measuring the capsid protein content and expressing the mTOR show that compared with the HCD group, the capsid protein content and the mTOR expressing amount of the HCB group are obviously increased.
Note: in fig. 9 "+" represents the difference between groups, and: p < 0.05, x: p < 0.01, x: p is less than 0.001, and the HCB group data in (a) - (c) are the average values of three groups of data, namely HCB1, HCB2 and HCB 3.
Therefore, the addition of the bacillus amyloliquefaciens SS1 can activate protein synthesis pathways by increasing the expression level of protein synthesis related genes, such as mTOR, further improve the total protein synthesis amount and increase the body protein deposition.
(5) Bacillus amyloliquefaciens SS1 for improving disease resistance of fish
1) Challenge test
Inoculating Aeromonas hydrophila CS4(A.hyd CS4) into L B liquid culture medium for resuscitation, shaking at 28 deg.C and 220rpm/min for 12h, inoculating resuscitation bacteria liquid into L B liquid culture medium at a ratio of 1:100 for amplification culture, shaking at 28 deg.C and 220rpm/min for 24h, centrifuging at 4 deg.C and 12000rpm/min for 20min, collecting thallus, re-suspending with PBS buffer solution, and adjusting concentration to 109CFU/m L, taking 21 fishes (such as tilapia) per group, and performing challenge experiment, wherein each fish is injected into abdominal cavity with 10 injections6CFU/g, death was observed and recorded for 7 days, and survival was calculated.
2) Breath burst test
Collecting 5 fish per group, anesthetizing with MS-222, separating fish body head kidney macrophage, separating head kidney macrophage by Percoll density gradient method, detecting and counting cell activity with trypan blue to ensure that the number of living cells is more than 90%, and adjusting cell concentration to 1 × 107cell/m L measurement of oxygen respiratory burst activity of head and kidney macrophages absorbance was measured at 540nm using NBT method, the results are shown in FIG. 10.
The results show that the survival rate of the HCD group is lowest, the survival rate of the HCB group is obviously increased relative to that of the HCD group by adding SS1, and the survival rate can be basically recovered to be the same as that of the CON group after 7 d; the oxygen respiration burst result of the macrophages of the head and kidney shows that the inhalation burst activity of the CON group and the inhalation burst activity of the HCD group have no obvious difference, and compared with the HCD group, the inhalation burst activity of the HCB group is obviously increased.
Note: in fig. 10 "+" represents the difference between groups, and: p < 0.05, x: p < 0.01, x: p is less than 0.001, and the HCB group data in (a) - (b) are the average values of HCB1, HCB2 and HCB 3.
Therefore, the bacillus amyloliquefaciens SS1 can effectively improve the activity of immune cells of fish bodies, promote the infection capacity of the fish against virus and germs (such as aeromonas hydrophila), enhance the comprehensive immunity and disease resistance of the fish bodies and further improve the survival rate.
In conclusion, the bacillus amyloliquefaciens SS1 separated by the method can be used as a safety functional feed additive to be added into high-starch fish feed, so that the utilization efficiency of carbohydrates in fish is improved, and the enzyme activity of digestive enzymes is improved by the bacillus amyloliquefaciens SS1 to promote growth; further, the Bacillus amyloliquefaciens SS1 can increase glycolysis level of liver of fish, thereby improving blood sugar steady state, relieving abnormal deposition of fat of fish by increasing energy consumption, and increasing body protein deposition by activating protein synthesis pathway; in addition, the bacillus amyloliquefaciens SS1 also has bacteriostatic property and can improve the disease resistance of fish bodies by protecting the immune system of organisms.
The technical features of the above embodiments 1 to 3 can be combined arbitrarily, and the combined technical solutions all belong to the protection scope of the present application. The foregoing shows and describes the general principles, essential features, and advantages of the invention. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, which are merely illustrative of the principles of the invention, but that various changes and modifications may be made without departing from the spirit and scope of the invention, which fall within the scope of the invention as claimed. The scope of the invention is defined by the appended claims and equivalents thereof.
Claims (10)
1. The Bacillus amyloliquefaciens is characterized by being Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) SS1 and being deposited in China center for type culture collection with the deposit numbers as follows: CCTCC M2020024, and the preservation date is 2020, 2 and 18.
2. Use of the bacillus amyloliquefaciens SS1 of claim 1 in the preparation of a bacteriostatic formulation.
3. Use of the bacillus amyloliquefaciens SS1 according to claim 1 for promoting fish growth.
4. Use of the bacillus amyloliquefaciens SS1 according to claim 1 for improving blood glucose homeostasis in fish.
5. Use of the bacillus amyloliquefaciens SS1 according to claim 1 for reducing fat deposits in fish.
6. Use of the bacillus amyloliquefaciens SS1 according to claim 1 for promoting the deposition of fish body proteins.
7. The use of the bacillus amyloliquefaciens SS1 as claimed in claim 1 for improving the disease resistance of fish.
8. An aquaculture feed comprising bacillus amyloliquefaciens SS1 of claim 1.
9. The aquaculture feed product of claim 10 wherein said bacillus amyloliquefaciens SS1 is present in said aquaculture feed product at a concentration of 105-107CFU/g。
10. A method of aquaculture wherein fish are fed with an aquaculture feed as claimed in claims 8 to 9 1 to 2 times per day at a rate of 4% by weight for a period of 5 to 10 weeks.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010109464.1A CN111394275B (en) | 2020-02-22 | 2020-02-22 | Bacillus amyloliquefaciens and application thereof, aquatic feed and aquaculture method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010109464.1A CN111394275B (en) | 2020-02-22 | 2020-02-22 | Bacillus amyloliquefaciens and application thereof, aquatic feed and aquaculture method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111394275A true CN111394275A (en) | 2020-07-10 |
| CN111394275B CN111394275B (en) | 2022-05-27 |
Family
ID=71427030
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010109464.1A Active CN111394275B (en) | 2020-02-22 | 2020-02-22 | Bacillus amyloliquefaciens and application thereof, aquatic feed and aquaculture method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111394275B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111763644A (en) * | 2020-07-24 | 2020-10-13 | 天津开发区坤禾生物技术有限公司 | Compound microbial inoculum for preventing and treating fish liver and gall syndrome as well as preparation method and application thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101418276A (en) * | 2008-12-08 | 2009-04-29 | 江南大学 | Host cell and method for efficient expression and secretion thereof in recombinant protein |
| CN103966118A (en) * | 2013-12-20 | 2014-08-06 | 中国水产科学研究院黄海水产研究所 | Bacillusamyloliquefaciens and application thereof |
| CN104651264A (en) * | 2014-12-22 | 2015-05-27 | 宝鸡市广仁生物科技有限公司 | Bacillus amyloliquefaciens and application thereof |
| EP3024479A2 (en) * | 2013-07-22 | 2016-06-01 | Aptalis Pharma Ltd. | High potency pancreatin pharmaceutical compositions |
| CN106434537A (en) * | 2016-09-13 | 2017-02-22 | 华东师范大学 | Method for culturing and inducing tilapia mossambica peritoneal preadipocytes and culture medium thereof |
| WO2019014118A1 (en) * | 2017-07-09 | 2019-01-17 | Igc Bio, Inc. | Pross optimized enzymes |
-
2020
- 2020-02-22 CN CN202010109464.1A patent/CN111394275B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101418276A (en) * | 2008-12-08 | 2009-04-29 | 江南大学 | Host cell and method for efficient expression and secretion thereof in recombinant protein |
| EP3024479A2 (en) * | 2013-07-22 | 2016-06-01 | Aptalis Pharma Ltd. | High potency pancreatin pharmaceutical compositions |
| CN103966118A (en) * | 2013-12-20 | 2014-08-06 | 中国水产科学研究院黄海水产研究所 | Bacillusamyloliquefaciens and application thereof |
| CN104651264A (en) * | 2014-12-22 | 2015-05-27 | 宝鸡市广仁生物科技有限公司 | Bacillus amyloliquefaciens and application thereof |
| CN106434537A (en) * | 2016-09-13 | 2017-02-22 | 华东师范大学 | Method for culturing and inducing tilapia mossambica peritoneal preadipocytes and culture medium thereof |
| WO2019014118A1 (en) * | 2017-07-09 | 2019-01-17 | Igc Bio, Inc. | Pross optimized enzymes |
Non-Patent Citations (4)
| Title |
|---|
| AMENY FARHAT-KHEMAKHEM等: "Assessment of the potential of the multi-enzyme producer bacillus amyloliquefaciens US573 as alternative feed additive", 《J SCI FOOD AGRIC》 * |
| RONG XU等: "Bacillus amyloliquefaciens ameliorates high-carbohydrate diet-induced metabolic phenotypes by restoration of intestinal acetate-producing bacteria in Nile Tilapia", 《BR J NUTR》 * |
| 吴树坤等: "沉香型酒醅中产香芽孢杆菌的分离鉴定及代谢产物分析", 《中国酿造》 * |
| 徐海燕等: "7株芽孢杆菌固体发酵条件下的产酶情况及对饲料消化能力的影响", 《青岛农业大学学报(自然科学版)》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111763644A (en) * | 2020-07-24 | 2020-10-13 | 天津开发区坤禾生物技术有限公司 | Compound microbial inoculum for preventing and treating fish liver and gall syndrome as well as preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111394275B (en) | 2022-05-27 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110272842A (en) | One plant of lactobacillus plantarum LP104 with fat reducing and weight reducing function | |
| CN102409015A (en) | Composite microecological preparation, application thereof in feed additive and premix | |
| CN105602997B (en) | Enterobacter cloacae prepare method and the application of biological nano elemental selenium | |
| CN110218681A (en) | One plant of lactobacillus fermenti KP101 and its application | |
| CN111304119B (en) | Feeding bacillus subtilis for degrading fumonisins and application thereof | |
| CN101801212B (en) | Use of cordyceps species or extract thereof in the egg production | |
| CN114574406B (en) | Lactobacillus rhamnosus strain WKA55, and application and product thereof in preparation of product for preventing and treating alcoholic liver injury | |
| CN101392223A (en) | Breeding method of microbial feed additive strain | |
| CN115322932B (en) | Lactobacillus plantarum with anti-alcohol and sobering-up capabilities and application thereof | |
| CN108531408A (en) | One plant of rhodotorula mucilaginosa new strains and probiotics | |
| CN102899276A (en) | Streptococcus thermophilus capable of lowering cholesterol levels and application thereof | |
| CN116769658A (en) | Bacillus coagulans AP15 and application thereof in intestinal tract regulation and uric acid and cholesterol metabolism | |
| CN115895966A (en) | Bifidobacterium bifidum BL002 for assisting in relieving gout and application thereof | |
| CN111394275B (en) | Bacillus amyloliquefaciens and application thereof, aquatic feed and aquaculture method | |
| CN113797232A (en) | Composition with function of relieving insulin resistance and application thereof | |
| CN111647525B (en) | Composite probiotics, microbial inoculum and application of composite probiotics and microbial inoculum in preparation of microbial inoculum for preventing and treating candida albicans vaginitis | |
| CN117586908A (en) | Lactobacillus plantarum LP15 strain capable of improving bone mineral density and preparation method and application thereof | |
| CN117660218A (en) | Lactobacillus reuteri LR108 strain capable of improving obesity of dogs and cats and application thereof | |
| CN114468150B (en) | Application of gentisic acid in promoting growth and rumen development of young ruminants | |
| CN106635834B (en) | The Thelephora ganbajun mycelium zinc polysaccharide and application that one plant of wizened bacteria strain and its fermentation obtain | |
| CN102851334A (en) | Fermentation medium and fermentation method of aflatoxin B1 | |
| CN109161501B (en) | Feeding bacillus licheniformis and application thereof | |
| CN110012989B (en) | Culture medium for prolonging life of drosophila melanogaster and preparation method and application method thereof | |
| CN116162557B (en) | A strain of Saccharomyces cerevisiae, chinese medicinal microecological preparation for preventing and treating ruminant diarrhea, and its preparation method | |
| CN111670999B (en) | Micro-ecological composite additive for aquatic products and application 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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |