CN113717881A - 一种凝结芽孢杆菌bc66及其应用 - Google Patents
一种凝结芽孢杆菌bc66及其应用 Download PDFInfo
- Publication number
- CN113717881A CN113717881A CN202110906213.0A CN202110906213A CN113717881A CN 113717881 A CN113717881 A CN 113717881A CN 202110906213 A CN202110906213 A CN 202110906213A CN 113717881 A CN113717881 A CN 113717881A
- Authority
- CN
- China
- Prior art keywords
- bacillus coagulans
- feed
- powder
- strain
- bacillus
- 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
- 241000193749 Bacillus coagulans Species 0.000 title claims abstract description 74
- 229940054340 bacillus coagulans Drugs 0.000 title claims abstract description 73
- 239000000843 powder Substances 0.000 claims abstract description 26
- 230000001580 bacterial effect Effects 0.000 claims abstract description 17
- 239000003814 drug Substances 0.000 claims abstract description 15
- 238000000855 fermentation Methods 0.000 claims abstract description 12
- 230000004151 fermentation Effects 0.000 claims abstract description 12
- 241001465754 Metazoa Species 0.000 claims abstract description 10
- 239000000654 additive Substances 0.000 claims abstract description 5
- 230000000996 additive effect Effects 0.000 claims abstract description 4
- 244000005700 microbiome Species 0.000 claims abstract description 4
- 241000287828 Gallus gallus Species 0.000 claims description 20
- 238000012360 testing method Methods 0.000 claims description 20
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 19
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 12
- 238000009360 aquaculture Methods 0.000 claims description 11
- 244000144974 aquaculture Species 0.000 claims description 11
- 239000007788 liquid Substances 0.000 claims description 11
- 244000144972 livestock Species 0.000 claims description 10
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 9
- 239000011780 sodium chloride Substances 0.000 claims description 9
- 238000002360 preparation method Methods 0.000 claims description 8
- 230000001954 sterilising effect Effects 0.000 claims description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 7
- 238000009395 breeding Methods 0.000 claims description 6
- 230000001488 breeding effect Effects 0.000 claims description 6
- 229940079593 drug Drugs 0.000 claims description 6
- 238000000034 method Methods 0.000 claims description 6
- 238000004321 preservation Methods 0.000 claims description 6
- 239000001888 Peptone Substances 0.000 claims description 4
- 108010080698 Peptones Proteins 0.000 claims description 4
- VTYYLEPIZMXCLO-UHFFFAOYSA-L calcium carbonate Substances [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims description 4
- 229910000019 calcium carbonate Inorganic materials 0.000 claims description 4
- 229940041514 candida albicans extract Drugs 0.000 claims description 4
- 229910000396 dipotassium phosphate Inorganic materials 0.000 claims description 4
- 239000008103 glucose Substances 0.000 claims description 4
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 4
- 229910000357 manganese(II) sulfate Inorganic materials 0.000 claims description 4
- 229910052603 melanterite Inorganic materials 0.000 claims description 4
- 235000019319 peptone Nutrition 0.000 claims description 4
- 239000012138 yeast extract Substances 0.000 claims description 4
- 235000013330 chicken meat Nutrition 0.000 claims description 3
- 238000009629 microbiological culture Methods 0.000 claims description 3
- 230000008569 process Effects 0.000 claims description 3
- 108090000623 proteins and genes Proteins 0.000 claims description 3
- 241000283690 Bos taurus Species 0.000 claims description 2
- 241000283973 Oryctolagus cuniculus Species 0.000 claims description 2
- 241001494479 Pecora Species 0.000 claims description 2
- 230000004913 activation Effects 0.000 claims description 2
- 229910052564 epsomite Inorganic materials 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims description 2
- 239000010802 sludge Substances 0.000 claims description 2
- 238000001694 spray drying Methods 0.000 claims description 2
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims 2
- 241000272517 Anseriformes Species 0.000 claims 1
- 241000282887 Suidae Species 0.000 claims 1
- 238000011081 inoculation Methods 0.000 claims 1
- 238000012807 shake-flask culturing Methods 0.000 claims 1
- 210000001035 gastrointestinal tract Anatomy 0.000 abstract description 14
- 244000052616 bacterial pathogen Species 0.000 abstract description 10
- 239000006041 probiotic Substances 0.000 abstract description 10
- 235000018291 probiotics Nutrition 0.000 abstract description 10
- 230000004083 survival effect Effects 0.000 abstract description 10
- 230000000529 probiotic effect Effects 0.000 abstract description 8
- 239000002253 acid Substances 0.000 abstract description 6
- 208000035240 Disease Resistance Diseases 0.000 abstract description 3
- 230000002401 inhibitory effect Effects 0.000 abstract description 3
- 241000193830 Bacillus <bacterium> Species 0.000 abstract description 2
- 239000003513 alkali Substances 0.000 abstract description 2
- 210000000941 bile Anatomy 0.000 abstract description 2
- 210000002429 large intestine Anatomy 0.000 abstract description 2
- 241000894006 Bacteria Species 0.000 description 11
- 230000002354 daily effect Effects 0.000 description 11
- 238000002474 experimental method Methods 0.000 description 11
- 230000000968 intestinal effect Effects 0.000 description 11
- 206010012735 Diarrhoea Diseases 0.000 description 10
- 241000607142 Salmonella Species 0.000 description 7
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 7
- SDOFMBGMRVAJNF-SLPGGIOYSA-N (2r,3r,4r,5s)-6-aminohexane-1,2,3,4,5-pentol Chemical compound NC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO SDOFMBGMRVAJNF-SLPGGIOYSA-N 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- 239000001963 growth medium Substances 0.000 description 6
- 241000251468 Actinopterygii Species 0.000 description 5
- 239000003833 bile salt Substances 0.000 description 5
- 210000004027 cell Anatomy 0.000 description 5
- 230000036039 immunity Effects 0.000 description 5
- 239000000725 suspension Substances 0.000 description 5
- 241000588724 Escherichia coli Species 0.000 description 4
- 241001138501 Salmonella enterica Species 0.000 description 4
- 241000193985 Streptococcus agalactiae Species 0.000 description 4
- 239000007853 buffer solution Substances 0.000 description 4
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 4
- 235000021050 feed intake Nutrition 0.000 description 4
- -1 gentiobicistine Chemical compound 0.000 description 4
- 238000011534 incubation Methods 0.000 description 4
- 230000005764 inhibitory process Effects 0.000 description 4
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 4
- 241000607522 Aeromonas sobria Species 0.000 description 3
- 241000193468 Clostridium perfringens Species 0.000 description 3
- 108060003951 Immunoglobulin Proteins 0.000 description 3
- 102000003777 Interleukin-1 beta Human genes 0.000 description 3
- 108090000193 Interleukin-1 beta Proteins 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 241000191967 Staphylococcus aureus Species 0.000 description 3
- 241000376029 Tachysurus fulvidraco Species 0.000 description 3
- 241000607272 Vibrio parahaemolyticus Species 0.000 description 3
- 230000003385 bacteriostatic effect Effects 0.000 description 3
- 235000014590 basal diet Nutrition 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 230000018109 developmental process Effects 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 230000003203 everyday effect Effects 0.000 description 3
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 3
- 210000004051 gastric juice Anatomy 0.000 description 3
- 102000018358 immunoglobulin Human genes 0.000 description 3
- 230000002757 inflammatory effect Effects 0.000 description 3
- 230000000366 juvenile effect Effects 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 230000001105 regulatory effect Effects 0.000 description 3
- 238000012163 sequencing technique Methods 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- WQZGKKKJIJFFOK-SVZMEOIVSA-N (+)-Galactose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-SVZMEOIVSA-N 0.000 description 2
- BJEPYKJPYRNKOW-REOHCLBHSA-N (S)-malic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O BJEPYKJPYRNKOW-REOHCLBHSA-N 0.000 description 2
- 108020004465 16S ribosomal RNA Proteins 0.000 description 2
- 238000009631 Broth culture Methods 0.000 description 2
- 206010010774 Constipation Diseases 0.000 description 2
- RFSUNEUAIZKAJO-VRPWFDPXSA-N D-Fructose Natural products OC[C@H]1OC(O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-VRPWFDPXSA-N 0.000 description 2
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 2
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 description 2
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 2
- JVTAAEKCZFNVCJ-REOHCLBHSA-N L-lactic acid Chemical compound C[C@H](O)C(O)=O JVTAAEKCZFNVCJ-REOHCLBHSA-N 0.000 description 2
- PNNNRSAQSRJVSB-UHFFFAOYSA-N L-rhamnose Natural products CC(O)C(O)C(O)C(O)C=O PNNNRSAQSRJVSB-UHFFFAOYSA-N 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- 230000002708 enhancing effect Effects 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 239000012634 fragment Substances 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 210000003736 gastrointestinal content Anatomy 0.000 description 2
- 230000003053 immunization Effects 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 230000003871 intestinal function Effects 0.000 description 2
- 239000004310 lactic acid Substances 0.000 description 2
- 235000014655 lactic acid Nutrition 0.000 description 2
- KWGKDLIKAYFUFQ-UHFFFAOYSA-M lithium chloride Chemical compound [Li+].[Cl-] KWGKDLIKAYFUFQ-UHFFFAOYSA-M 0.000 description 2
- 239000002207 metabolite Substances 0.000 description 2
- 239000012982 microporous membrane Substances 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 150000003536 tetrazoles Chemical class 0.000 description 2
- 230000004584 weight gain Effects 0.000 description 2
- 235000019786 weight gain Nutrition 0.000 description 2
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 description 1
- OGLCQHRZUSEXNB-WHDMSYDLSA-N (3r,3ar,5r,6r,6ar)-3,5,6-trihydroxy-hexahydrofuro[3,2-b]furan-2-one Chemical compound O[C@H]1C(=O)O[C@@H]2[C@@H](O)[C@H](O)O[C@@H]21 OGLCQHRZUSEXNB-WHDMSYDLSA-N 0.000 description 1
- CYDQOEWLBCCFJZ-UHFFFAOYSA-N 4-(4-fluorophenyl)oxane-4-carboxylic acid Chemical compound C=1C=C(F)C=CC=1C1(C(=O)O)CCOCC1 CYDQOEWLBCCFJZ-UHFFFAOYSA-N 0.000 description 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- 241000272525 Anas platyrhynchos Species 0.000 description 1
- WZPBZJONDBGPKJ-UHFFFAOYSA-N Antibiotic SQ 26917 Natural products O=C1N(S(O)(=O)=O)C(C)C1NC(=O)C(=NOC(C)(C)C(O)=O)C1=CSC(N)=N1 WZPBZJONDBGPKJ-UHFFFAOYSA-N 0.000 description 1
- FRYULLIZUDQONW-IMJSIDKUSA-N Asp-Asp Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(O)=O FRYULLIZUDQONW-IMJSIDKUSA-N 0.000 description 1
- 102000016938 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- HEBKCHPVOIAQTA-NGQZWQHPSA-N D-Arabitol Natural products OC[C@H](O)C(O)[C@H](O)CO HEBKCHPVOIAQTA-NGQZWQHPSA-N 0.000 description 1
- GUBGYTABKSRVRQ-UHFFFAOYSA-N D-Cellobiose Natural products OCC1OC(OC2C(O)C(O)C(O)OC2CO)C(O)C(O)C1O GUBGYTABKSRVRQ-UHFFFAOYSA-N 0.000 description 1
- GUBGYTABKSRVRQ-CUHNMECISA-N D-Cellobiose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-CUHNMECISA-N 0.000 description 1
- GSXOAOHZAIYLCY-UHFFFAOYSA-N D-F6P Natural products OCC(=O)C(O)C(O)C(O)COP(O)(O)=O GSXOAOHZAIYLCY-UHFFFAOYSA-N 0.000 description 1
- AEMOLEFTQBMNLQ-DTEWXJGMSA-N D-Galacturonic acid Natural products O[C@@H]1O[C@H](C(O)=O)[C@H](O)[C@H](O)[C@H]1O AEMOLEFTQBMNLQ-DTEWXJGMSA-N 0.000 description 1
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 1
- MTCFGRXMJLQNBG-UWTATZPHSA-N D-Serine Chemical compound OC[C@@H](N)C(O)=O MTCFGRXMJLQNBG-UWTATZPHSA-N 0.000 description 1
- 229930195711 D-Serine Natural products 0.000 description 1
- HEBKCHPVOIAQTA-QWWZWVQMSA-N D-arabinitol Chemical compound OC[C@@H](O)C(O)[C@H](O)CO HEBKCHPVOIAQTA-QWWZWVQMSA-N 0.000 description 1
- SHZGCJCMOBCMKK-SVZMEOIVSA-N D-fucopyranose Chemical compound C[C@H]1OC(O)[C@H](O)[C@@H](O)[C@H]1O SHZGCJCMOBCMKK-SVZMEOIVSA-N 0.000 description 1
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- RGHNJXZEOKUKBD-SQOUGZDYSA-N D-gluconic acid Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C(O)=O RGHNJXZEOKUKBD-SQOUGZDYSA-N 0.000 description 1
- RGHNJXZEOKUKBD-UHFFFAOYSA-N D-gluconic acid Natural products OCC(O)C(O)C(O)C(O)C(O)=O RGHNJXZEOKUKBD-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 1
- 238000007400 DNA extraction Methods 0.000 description 1
- 239000004375 Dextrin Substances 0.000 description 1
- 229920001353 Dextrin Polymers 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- IECPWNUMDGFDKC-UHFFFAOYSA-N Fusicsaeure Natural products C12C(O)CC3C(=C(CCC=C(C)C)C(O)=O)C(OC(C)=O)CC3(C)C1(C)CCC1C2(C)CCC(O)C1C IECPWNUMDGFDKC-UHFFFAOYSA-N 0.000 description 1
- IAJILQKETJEXLJ-UHFFFAOYSA-N Galacturonsaeure Natural products O=CC(O)C(O)C(O)C(O)C(O)=O IAJILQKETJEXLJ-UHFFFAOYSA-N 0.000 description 1
- 241000282414 Homo sapiens Species 0.000 description 1
- OJMMVQQUTAEWLP-UHFFFAOYSA-N Lincomycin Natural products CN1CC(CCC)CC1C(=O)NC(C(C)O)C1C(O)C(O)C(O)C(SC)O1 OJMMVQQUTAEWLP-UHFFFAOYSA-N 0.000 description 1
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 1
- 206010028116 Mucosal inflammation Diseases 0.000 description 1
- 201000010927 Mucositis Diseases 0.000 description 1
- OVRNDRQMDRJTHS-UHFFFAOYSA-N N-acelyl-D-glucosamine Natural products CC(=O)NC1C(O)OC(CO)C(O)C1O OVRNDRQMDRJTHS-UHFFFAOYSA-N 0.000 description 1
- OVRNDRQMDRJTHS-FMDGEEDCSA-N N-acetyl-beta-D-glucosamine Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O OVRNDRQMDRJTHS-FMDGEEDCSA-N 0.000 description 1
- OVRNDRQMDRJTHS-OZRXBMAMSA-N N-acetyl-beta-D-mannosamine Chemical compound CC(=O)N[C@@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O OVRNDRQMDRJTHS-OZRXBMAMSA-N 0.000 description 1
- DKXNBNKWCZZMJT-UHFFFAOYSA-N O4-alpha-D-Mannopyranosyl-D-mannose Natural products O=CC(O)C(O)C(C(O)CO)OC1OC(CO)C(O)C(O)C1O DKXNBNKWCZZMJT-UHFFFAOYSA-N 0.000 description 1
- RZPAKFUAFGMUPI-UHFFFAOYSA-N Oleandomycin Natural products O1C(C)C(O)C(OC)CC1OC1C(C)C(=O)OC(C)C(C)C(O)C(C)C(=O)C2(OC2)CC(C)C(OC2C(C(CC(C)O2)N(C)C)O)C1C RZPAKFUAFGMUPI-UHFFFAOYSA-N 0.000 description 1
- 239000004104 Oleandomycin Substances 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 102000057297 Pepsin A Human genes 0.000 description 1
- 108090000284 Pepsin A Proteins 0.000 description 1
- HJYYPODYNSCCOU-ZDHWWVNNSA-N Rifamycin SV Natural products COC1C=COC2(C)Oc3c(C)c(O)c4c(O)c(NC(=O)C(=C/C=C/C(C)C(O)C(C)C(O)C(C)C(OC(=O)C)C1C)C)cc(O)c4c3C2=O HJYYPODYNSCCOU-ZDHWWVNNSA-N 0.000 description 1
- 241000282898 Sus scrofa Species 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 108010059993 Vancomycin Proteins 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 229940022663 acetate Drugs 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- IAJILQKETJEXLJ-RSJOWCBRSA-N aldehydo-D-galacturonic acid Chemical compound O=C[C@H](O)[C@@H](O)[C@@H](O)[C@H](O)C(O)=O IAJILQKETJEXLJ-RSJOWCBRSA-N 0.000 description 1
- PNNNRSAQSRJVSB-BXKVDMCESA-N aldehydo-L-rhamnose Chemical compound C[C@H](O)[C@H](O)[C@@H](O)[C@@H](O)C=O PNNNRSAQSRJVSB-BXKVDMCESA-N 0.000 description 1
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 description 1
- WQZGKKKJIJFFOK-DVKNGEFBSA-N alpha-D-glucose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-DVKNGEFBSA-N 0.000 description 1
- BJEPYKJPYRNKOW-UHFFFAOYSA-N alpha-hydroxysuccinic acid Natural products OC(=O)C(O)CC(O)=O BJEPYKJPYRNKOW-UHFFFAOYSA-N 0.000 description 1
- 229940024606 amino acid Drugs 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 239000003674 animal food additive Substances 0.000 description 1
- 230000003042 antagnostic effect Effects 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- WZPBZJONDBGPKJ-VEHQQRBSSA-N aztreonam Chemical compound O=C1N(S([O-])(=O)=O)[C@@H](C)[C@@H]1NC(=O)C(=N/OC(C)(C)C(O)=O)\C1=CSC([NH3+])=N1 WZPBZJONDBGPKJ-VEHQQRBSSA-N 0.000 description 1
- 229960003644 aztreonam Drugs 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- AEMOLEFTQBMNLQ-UHFFFAOYSA-N beta-D-galactopyranuronic acid Natural products OC1OC(C(O)=O)C(O)C(O)C1O AEMOLEFTQBMNLQ-UHFFFAOYSA-N 0.000 description 1
- VOIFKEWOFUNPBN-QIUUJYRFSA-N beta-D-glucuronamide Chemical compound NC(=O)[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O VOIFKEWOFUNPBN-QIUUJYRFSA-N 0.000 description 1
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 235000019425 dextrin Nutrition 0.000 description 1
- 235000019621 digestibility Nutrition 0.000 description 1
- 102000038379 digestive enzymes Human genes 0.000 description 1
- 108091007734 digestive enzymes Proteins 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- 239000003640 drug residue Substances 0.000 description 1
- 230000000459 effect on growth Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 229960004675 fusidic acid Drugs 0.000 description 1
- IECPWNUMDGFDKC-MZJAQBGESA-N fusidic acid Chemical compound O[C@@H]([C@@H]12)C[C@H]3\C(=C(/CCC=C(C)C)C(O)=O)[C@@H](OC(C)=O)C[C@]3(C)[C@@]2(C)CC[C@@H]2[C@]1(C)CC[C@@H](O)[C@H]2C IECPWNUMDGFDKC-MZJAQBGESA-N 0.000 description 1
- 230000002496 gastric effect Effects 0.000 description 1
- 235000012208 gluconic acid Nutrition 0.000 description 1
- 229950006191 gluconic acid Drugs 0.000 description 1
- 125000002791 glucosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 description 1
- 229950004542 glucuronamide Drugs 0.000 description 1
- 229960000789 guanidine hydrochloride Drugs 0.000 description 1
- PJJJBBJSCAKJQF-UHFFFAOYSA-N guanidinium chloride Chemical compound [Cl-].NC(N)=[NH2+] PJJJBBJSCAKJQF-UHFFFAOYSA-N 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 238000002649 immunization Methods 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 210000002490 intestinal epithelial cell Anatomy 0.000 description 1
- 230000007413 intestinal health Effects 0.000 description 1
- 210000004731 jugular vein Anatomy 0.000 description 1
- GSXOAOHZAIYLCY-HSUXUTPPSA-N keto-D-fructose 6-phosphate Chemical compound OCC(=O)[C@@H](O)[C@H](O)[C@H](O)COP(O)(O)=O GSXOAOHZAIYLCY-HSUXUTPPSA-N 0.000 description 1
- 229940116298 l- malic acid Drugs 0.000 description 1
- 229960005287 lincomycin Drugs 0.000 description 1
- OJMMVQQUTAEWLP-KIDUDLJLSA-N lincomycin Chemical compound CN1C[C@H](CCC)C[C@H]1C(=O)N[C@H]([C@@H](C)O)[C@@H]1[C@H](O)[C@H](O)[C@@H](O)[C@@H](SC)O1 OJMMVQQUTAEWLP-KIDUDLJLSA-N 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 235000011090 malic acid Nutrition 0.000 description 1
- 229960002160 maltose Drugs 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 235000012054 meals Nutrition 0.000 description 1
- DLRVVLDZNNYCBX-ABXHMFFYSA-N melibiose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)C(O)O1 DLRVVLDZNNYCBX-ABXHMFFYSA-N 0.000 description 1
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- HOVAGTYPODGVJG-XUUWZHRGSA-N methyl beta-D-glucopyranoside Chemical compound CO[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O HOVAGTYPODGVJG-XUUWZHRGSA-N 0.000 description 1
- CWKLZLBVOJRSOM-UHFFFAOYSA-N methyl pyruvate Chemical compound COC(=O)C(C)=O CWKLZLBVOJRSOM-UHFFFAOYSA-N 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 230000003278 mimic effect Effects 0.000 description 1
- DYKFCLLONBREIL-KVUCHLLUSA-N minocycline Chemical compound C([C@H]1C2)C3=C(N(C)C)C=CC(O)=C3C(=O)C1=C(O)[C@@]1(O)[C@@H]2[C@H](N(C)C)C(O)=C(C(N)=O)C1=O DYKFCLLONBREIL-KVUCHLLUSA-N 0.000 description 1
- 229960004023 minocycline Drugs 0.000 description 1
- 210000004400 mucous membrane Anatomy 0.000 description 1
- 229950006780 n-acetylglucosamine Drugs 0.000 description 1
- MHWLWQUZZRMNGJ-UHFFFAOYSA-N nalidixic acid Chemical compound C1=C(C)N=C2N(CC)C=C(C(O)=O)C(=O)C2=C1 MHWLWQUZZRMNGJ-UHFFFAOYSA-N 0.000 description 1
- 229960000210 nalidixic acid Drugs 0.000 description 1
- 238000010899 nucleation Methods 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- RZPAKFUAFGMUPI-KGIGTXTPSA-N oleandomycin Chemical compound O1[C@@H](C)[C@H](O)[C@@H](OC)C[C@@H]1O[C@@H]1[C@@H](C)C(=O)O[C@H](C)[C@H](C)[C@H](O)[C@@H](C)C(=O)[C@]2(OC2)C[C@H](C)[C@H](O[C@H]2[C@@H]([C@H](C[C@@H](C)O2)N(C)C)O)[C@H]1C RZPAKFUAFGMUPI-KGIGTXTPSA-N 0.000 description 1
- 229960002351 oleandomycin Drugs 0.000 description 1
- 235000019367 oleandomycin Nutrition 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 229940111202 pepsin Drugs 0.000 description 1
- BFPJYWDBBLZXOM-UHFFFAOYSA-L potassium tellurite Chemical compound [K+].[K+].[O-][Te]([O-])=O BFPJYWDBBLZXOM-UHFFFAOYSA-L 0.000 description 1
- 229960002429 proline Drugs 0.000 description 1
- HJYYPODYNSCCOU-ODRIEIDWSA-N rifamycin SV Chemical compound OC1=C(C(O)=C2C)C3=C(O)C=C1NC(=O)\C(C)=C/C=C/[C@H](C)[C@H](O)[C@@H](C)[C@@H](O)[C@@H](C)[C@H](OC(C)=O)[C@H](C)[C@@H](OC)\C=C\O[C@@]1(C)OC2=C3C1=O HJYYPODYNSCCOU-ODRIEIDWSA-N 0.000 description 1
- 229940109171 rifamycin sv Drugs 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000011218 seed culture Methods 0.000 description 1
- 150000004666 short chain fatty acids Chemical class 0.000 description 1
- 235000021391 short chain fatty acids Nutrition 0.000 description 1
- 238000004088 simulation Methods 0.000 description 1
- XUXNAKZDHHEHPC-UHFFFAOYSA-M sodium bromate Chemical compound [Na+].[O-]Br(=O)=O XUXNAKZDHHEHPC-UHFFFAOYSA-M 0.000 description 1
- MFBOGIVSZKQAPD-UHFFFAOYSA-M sodium butyrate Chemical compound [Na+].CCCC([O-])=O MFBOGIVSZKQAPD-UHFFFAOYSA-M 0.000 description 1
- 239000001540 sodium lactate Substances 0.000 description 1
- 235000011088 sodium lactate Nutrition 0.000 description 1
- 229940005581 sodium lactate Drugs 0.000 description 1
- 229960000776 sodium tetradecyl sulfate Drugs 0.000 description 1
- GNBVPFITFYNRCN-UHFFFAOYSA-M sodium thioglycolate Chemical compound [Na+].[O-]C(=O)CS GNBVPFITFYNRCN-UHFFFAOYSA-M 0.000 description 1
- UPUIQOIQVMNQAP-UHFFFAOYSA-M sodium;tetradecyl sulfate Chemical compound [Na+].CCCCCCCCCCCCCCOS([O-])(=O)=O UPUIQOIQVMNQAP-UHFFFAOYSA-M 0.000 description 1
- 235000010356 sorbitol Nutrition 0.000 description 1
- 229960002920 sorbitol Drugs 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 230000028070 sporulation Effects 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 229960003165 vancomycin Drugs 0.000 description 1
- MYPYJXKWCTUITO-LYRMYLQWSA-N vancomycin Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=C2C=C3C=C1OC1=CC=C(C=C1Cl)[C@@H](O)[C@H](C(N[C@@H](CC(N)=O)C(=O)N[C@H]3C(=O)N[C@H]1C(=O)N[C@H](C(N[C@@H](C3=CC(O)=CC(O)=C3C=3C(O)=CC=C1C=3)C(O)=O)=O)[C@H](O)C1=CC=C(C(=C1)Cl)O2)=O)NC(=O)[C@@H](CC(C)C)NC)[C@H]1C[C@](C)(N)[C@H](O)[C@H](C)O1 MYPYJXKWCTUITO-LYRMYLQWSA-N 0.000 description 1
- MYPYJXKWCTUITO-UHFFFAOYSA-N vancomycin Natural products O1C(C(=C2)Cl)=CC=C2C(O)C(C(NC(C2=CC(O)=CC(O)=C2C=2C(O)=CC=C3C=2)C(O)=O)=O)NC(=O)C3NC(=O)C2NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(CC(C)C)NC)C(O)C(C=C3Cl)=CC=C3OC3=CC2=CC1=C3OC1OC(CO)C(O)C(O)C1OC1CC(C)(N)C(O)C(C)O1 MYPYJXKWCTUITO-UHFFFAOYSA-N 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Images
Classifications
-
- 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
- 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
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Microbiology (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- General Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Biochemistry (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Polymers & Plastics (AREA)
- Molecular Biology (AREA)
- Mycology (AREA)
- Animal Behavior & Ethology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biomedical Technology (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- General Engineering & Computer Science (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Physiology (AREA)
- Animal Husbandry (AREA)
- Epidemiology (AREA)
- Food Science & Technology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
本发明属于微生物技术领域，涉及一种凝结芽孢杆菌BC66及其应用。本发明凝结芽孢杆菌BC66菌株分离来源于仔猪肠道，产酸快，具有较强的抑制病原菌的能力，抗病能力强。芽孢数高，100L发酵罐放大培养制备的菌粉芽孢数可达到2.5×1011CFU/g。该菌株制备的菌粉能耐受酸碱和胆汁，顺利通过胃肠道环境进入大肠，凝结芽孢杆菌BC66的细胞存活率高达88.5%，能在机体内有效发挥益生作用。本发明提供所述的凝结芽孢杆菌BC66或其菌粉在畜牧养殖中作为饲料或药物的添加剂的应用。
Description
技术领域
本发明属于微生物技术领域，尤其是涉及一种益生凝结芽孢杆菌BC66及其应用。
背景技术
目前，我国在畜牧或水产养殖过程中由于过量使用抗生素、化学药物，以及激素等导致药物残留、污染问题，长期下来致使畜牧或水产类产品安全性降低，影响人类健康。同时在养殖成本上，比如动物出现抵抗力下降、肠道功能紊乱、疾病频发等影响，导致动物生长性能受到影响，疾病率和死亡率上升，成本较大。
凝结芽孢杆菌是一种不仅能产乳酸，还能产芽孢的革兰氏阳性细菌。凝结芽孢杆菌除了具有乳酸菌的所有特点外，还具有耐高温高压，抗逆性强，代谢旺盛和稳定的存储性等优良特性。凝结芽孢杆菌在肠道中还能产生大量的有益代谢产物，分泌凝结素、多种消化酶、氨基酸和短链脂肪酸等代谢产物，这些物质有助于机体消化吸收，抑制有害细菌生长，从而有效地调节肠道微生物菌群平衡，增加肠道上皮细胞的黏膜性，有利于有益菌在肠道中定植，提高上皮细胞的生存力，建立完整的生物屏障系统，提高机体的免疫力。
近年来，凝结芽孢杆菌作为一种新型饲用微生态制剂，能有效调节肠道微生态菌群平衡，增强机体免疫力，提高生长性能，防治疾病，在畜牧和水产养殖业中具有广泛应用前景。
发明内容
本发明的目的在于提供一种凝结芽孢杆菌BC66及其在畜牧和水产养殖领域的应用。
本发明所述技术方案如下：
一种凝结芽孢杆菌BC66，其微生物保藏号为CGMCC21879；分类命名为：Bacilluscoagulans；保藏时间：2021年03月08日；保藏地址：北京市朝阳区北辰西路1号院3号；保藏单位：中国微生物菌种保藏管理委员会普通微生物中心。
所述凝结芽孢杆菌BC66的16SrRNA基因序列如SEQ ID No 1所示。
所述凝结芽孢杆菌BC66分离于健康仔猪肠道内容物，具有产酸快特点。对大肠杆菌、金黄色葡萄球菌、猪霍乱沙门氏菌、产气夹膜梭菌、温和气单胞菌、副溶血性弧菌和无乳链球菌等畜牧或水产类病原菌具有显著的抑制作用，能增强机体抗病能力，改善机体肠道微生态环境。
本发明还提供所述凝结芽孢杆菌BC66的益生菌菌粉，菌粉芽孢数高，可达到2.5×1011CFU/g。
所述的凝结芽孢杆菌BC66益生菌菌粉是采用下述方法制备的：
凝结芽孢杆菌菌种从甘油管出发，接入试管斜面活化，经一级种子摇瓶(150ml/500mL)培养后，按照5％接种量转入种子罐(3L/5L)扩培，得到二级种子液；将二级种子液转入100L发酵罐(60L/100L)发酵，得到的发酵液经12000rmp离心后收集菌泥，通过喷雾干燥工艺得到菌粉。菌粉进行活菌计数检测，菌粉检测芽孢数可达到2.5×1011CFU/g。
上述菌粉制备过程中，所用的种子培养基配方以重量百分比计，葡萄糖2.0％、酵母浸粉1.0％、蛋白胨1.0％、MgSO4·7H2O 0.03％、MnSO4 0.01％、NaCl 0.05％、K2HPO40.1％、FeSO4·7H2O 0.001％，CaCO3 0.05％，余量为水，调节初始pH值为7.0±0.1，115℃灭菌30min备用。
上述菌粉制备过程中，所用的发酵培养基配方以重量百分比计，葡萄糖3.0％、酵母浸粉3.5％、蛋白胨4.0％、MgSO4·7H2O 0.06％、MnSO4 0.01％、NaCl 0.1％、K2HPO40.2％、FeSO4·7H2O 0.002％，CaCO3 0.1％，余量为水，调节初始pH值为7.0±0.1，115℃灭菌30min备用。
本发明所述的凝结芽孢杆菌BC66具有较强的通过胃肠道的能力。在模拟胃液pH2.5，肠液pH8和胆盐0.3wt％中分别连续处理2.5h后，BC66的细胞存活率还高达88.5％，保证大量活菌进入肠道，在机体内有效发挥益生作用。
本发明所述的凝结芽孢杆菌BC66还具有调节肠道菌群，增强肠道功能，促进生长发育，改善便秘，缓解腹泻以及提高机体免疫力。
特别地，本发明提供所述的凝结芽孢杆菌BC66或其菌粉在畜牧养殖中作为饲料或药物的添加剂的应用。
本发明提供所述的凝结芽孢杆菌BC66或其菌粉在制备畜牧养殖饲料或药物的添加剂中的应用。本发明还提供所述的凝结芽孢杆菌BC66或其菌粉在制备畜牧养殖饲料或药物中的应用。
所述的饲料或药物包括用于猪、鸡、鸭、牛、羊、兔等的饲料或药物。
所述的饲料也包括用于水产养殖的饲料或药物。
特别地，本发明提供所述的凝结芽孢杆菌BC66或其菌粉在水产养殖中的应用。
本发明提供所述的凝结芽孢杆菌BC66或其菌粉在制备水产养殖饲料或药物中的应用。
本发明凝结芽孢杆菌BC66菌株分离来源于仔猪肠道，产酸快，具有较强的抑制病原菌的能力，抗病能力强。芽孢数高，100L发酵罐放大培养制备的菌粉芽孢数可达到2.5×1011CFU/g。该菌株制备的菌粉能耐受酸碱和胆汁，顺利通过胃肠道环境进入大肠，凝结芽孢杆菌BC66的细胞存活率高达88.5％，能在机体内有效发挥益生作用。BC66还具有调节肠道菌群，增强肠道功能，促进生长发育，改善便秘，缓解腹泻，提高机体免疫力，可广泛应用于畜牧和水产养殖业领域。
附图说明
图1凝结芽孢杆菌BC66典型的芽孢形态。
图2凝结芽孢杆菌BC66对肠粘膜免疫球蛋白IgA的影响。
图3凝结芽孢杆菌BC66对肠粘膜炎症因子IL-1β的影响。
具体实施方式
下面结合实施例对本发明做进一步详细的描述，但本发明的实施方式不限于此。
实施例1凝结芽孢杆菌BC66的筛选
微康益生菌(苏州)股份有限公司保藏了两万余株来源于自然界分离的不同属种的益生菌菌株，其中有50株是来源于健康仔猪肠道内容物的凝结芽孢杆菌，我们从这些菌株中筛选到一株产酸能力特别强的菌株BC66。
实施例2凝结芽孢杆菌BC66理化特性
凝结芽孢杆菌BC66，革兰氏阳性杆状细菌，接触酶阳性，氧化酶阴性，可形成芽孢，芽孢端生。生长实验结果显示该菌株能利用α-D-葡萄糖、D-甘露糖、龙胆二塘、α-D-乳糖、D-果糖、D-蜜二糖、D-海藻糖、D-半乳糖、D-纤维二塘、D-岩藻糖、L-岩藻糖、L-鼠李糖、D-麦芽糖、松二塘、D-果糖-6-磷酸、甘氨酸-L-脯氨酸、D-半乳糖醛酸、D-葡糖酸、L-苹果酸、L-半乳糖酸内酯、丙酮酸甲酯、D-葡萄糖酸醛、D-甘露醇、D-阿糖醇、β-甲基-D-葡糖苷、N-乙酰-D-葡糖胺、葡糖醛酰胺、N-乙酰-β-D-甘露糖胺、糊精、甘油、L-乳酸。化学敏感实验结果显示，该菌株对醋竹桃霉素、利福霉素SV、二甲胺四环素、万古霉素、十四烷硫酸钠、四唑蓝、夫西地酸、林可霉素、盐酸胍、亚碲酸钾、溴酸钠、氯化锂、4％NaCl、8％NaCl敏感，D-丝氨酸、四唑紫、萘啶酸、氨曲南、丁酸纳、1％乳酸钠、1％NaCl不敏感，能够在pH6.0环境下生长。
实施例3凝结芽孢杆菌BC66的16S rRNA基因序列测定结果
液体扩培，收集菌体，提取基因组DNA，采用通用引物27F：AGAGTTTGATCCTGGCTCAG(SEQ ID No 2)和1492R：GGTTACCTTGTTACGACTT(SEQ ID No 3)扩增其16S rDNA片段，用琼脂糖凝胶电泳检测PCR扩增产物。将获得的目的片段进行测序分析，测序结果在NCBI数据库中进行比对分析，鉴定所得菌株为凝结芽孢杆菌，其分类命名为Bacillus coagulans，该菌株已于2021年03月08日保藏于中国微生物菌种保藏管理委员会普通微生物中心(CGMCC)，地址：北京市朝阳区北辰西路1号院3号，保藏编号为CGMCC21879，菌株编号为BC66。
实施例4凝结芽孢杆菌BC66抑制病原菌实验
(1)凝结芽孢杆菌BC66发酵液制备：将凝结芽孢杆菌BC66按一定比例接种至液体MRS培养基中，42℃培养24h，得到发酵液。
(2)指标菌菌液制备：指示菌株分别是大肠杆菌、金黄色葡萄球菌、猪霍乱沙门氏菌、产气夹膜梭菌、温和气单胞菌、副溶血性弧菌和无乳链球菌。将指示菌株0.1mL接种于10mL液体LB培养基中，于37℃220rpm恒温摇床培养24h。离心，收集菌体，用PBS缓冲液清洗2次，再次用PBS缓冲液充分重悬，调整病原菌浓度至108-1010CFU/mL。
具体地，将含有1.5％琼脂的MRS肉汤培养基冷却至55℃左右，与指示菌悬液按一定比例混匀，然后迅速倾注于预先放置牛津杯的平板中，待培养基冷却凝固后，取出牛津杯，在每孔注入200μL拮抗菌液，以200μL无菌MRS液体培养基作为空白对照，静置数分钟后，放入37℃恒温培养箱中，培养适宜时间后观察，用游标卡尺测量抑菌圈直径，每个菌进行三个重复，结果见表3。
表1凝结芽孢杆菌BC66对病原菌抑菌效果
| 病原菌 | 抑菌圈直径(mm) |
| 大肠杆菌 | 23±0.11 |
| 金黄色葡萄球菌 | 18±0.22 |
| 猪霍乱沙门氏菌 | 22±0.24 |
| 产气夹膜梭菌 | 12±0.48 |
| 温和气单胞菌 | 10±0.26 |
| 副溶血性弧菌 | 13±0.36 |
| 无乳链球菌 | 24±0.66 |
表2结果显示凝结芽孢杆菌BC66对多种病原菌均具有明显的抑制作用，其中对大肠杆菌、猪霍乱沙门氏菌和无乳链球菌的抑制作用极其显著。
实施例5凝结芽孢杆菌BC66体外模拟通过胃肠道的能力
(1)模拟人工胃液制备：配制0.5％氯化钠溶液，加入0.3％胃蛋白酶，用1mol/LHCL调节pH值至2.5，然后充分溶解后用0.22μm微孔滤膜过滤除菌备用。
(2)模拟人工肠液制备：配制0.5％NaCl溶液，加入0.1％胰蛋白酶，用0.1mol/LNaOH调节pH值至8.0，然后充分溶解后用0.22μm微孔滤膜过滤除菌备用。
(3)胆盐培养液：猪胆盐0.3％、巯基乙酸钠0.2％、MRS液体培养基，pH6.0～6.2，118℃，15min灭菌。
(4)体外模拟通过整个胃肠道的环境耐受性实验
称取0.1样品置于50mL离心管中，加入9.9ml模拟人工胃液，在37℃条件下孵育2.5h。孵育结束后，用PBS缓冲溶液冲洗，然后在5000rpm下离心3min，去掉上清液后，再加入9.9mL模拟人工肠液中，在37℃下继续孵育2.5h。孵育结束后，用PBS缓冲溶液冲洗，然后在5000rpm下离心3min，去掉上清液后，再加入9.9mL胆盐培养液，在37℃下继续孵育2.5h。模拟消化过程结束后收集测定活菌数，评价凝结芽孢杆菌BC66细胞的存活情况。其中，存活率％＝Nt/N0×100％，式中N0表示菌株未经耐受的活菌数(CFU/mL)，Nt表示经过胃肠道模拟耐受之后的活菌数(CFU/mL)。
检测结果表明，通过模拟胃液、肠液和胆盐整个操作流程，本发明的菌株BC66的存活率为88.5％。实验说明凝结芽孢杆菌BC66能顺利通过胃肠道耐受环境，有高达88.5％的活菌可以安全到达肠道中生长繁殖。
实施例6凝结芽孢杆菌BC66作为饲料添加剂对肉仔鸡性能的影响
试验动物选择健康、体重相近的1日龄AA肉仔鸡600只，经检验各实验动物体重差异不显著(±0.2kg)(P＞0.05)。随机分成4组，每组5个重复，每组30只，分别是对照组、试验1组、试验2组和试验3组。
对照组：饲喂基础日粮；
试验1组：饲喂基础日粮+凝结芽孢杆菌；
试验2组：饲喂基础日粮+沙门氏菌；
试验3组：饲喂基础日粮+凝结芽孢杆菌+沙门氏菌。
每个处理组饲喂基础日料，进行预饲期7天。开始正式试验，对照组每日每只罐服无菌生理盐水；试验1组每日每只罐服108CFU数量级的凝结芽孢杆菌BC66菌悬液；试验2组每日每只罐服108CFU数量级的沙门氏菌菌悬液；试验3组每日每只罐服108CFU数量级的凝结芽孢杆菌菌悬液，同时罐服108CFU数量级的沙门氏菌菌悬液；正式实验期为21天。试验前对鸡舍进行全面消毒，试验期间肉仔鸡全期采用笼养方式饲养，自然光照，自由采食、饮水，常规的日常管理，按正常免疫程序进行免疫接种。试验期间观察肉仔鸡的生长状况、并进行记录，统计各组平均日增重、平均日采食量、料重比和腹泻率。
表2凝结芽孢杆菌BC66对肉仔鸡生长性能的影响
注：同列数据肩标字母相同表示差异不显著(P＞0.05)，肩标字母不同小写字母表示差异显著(P＜0.05)。
结果表明，试验1组肉仔鸡在平均日增重和平均日采食都显著高于对照组，肉仔鸡腹泻率显著低于对照组；试验2组罐服有沙门氏菌病原菌，平均日增重和平均日采食量显著低于试验1组，肉仔鸡腹泻率显著高于试验1组。结果显示，肉仔鸡服用病原菌沙门氏菌后，阻碍了肉仔鸡的体重生长，也增加了肉仔鸡的腹泻率。服用凝结芽孢杆菌显著提升了肉仔鸡的体重和采食量，同时对腹泻也有明显缓解作用，提升肉仔鸡的存活率。试验3组平均日增重、平均日采食量均显著高于试验2组，肉仔鸡腹泻率显著降低。结果显示，服用凝结芽孢杆菌显著性地提升了肉仔鸡的体重，对沙门氏菌病原菌引起的生长阻碍和腹泻率增加均具有缓解作用。说明凝结芽孢杆菌BC66能明显提升肉仔鸡日增重，促进动物生长，缓解动物腹泻，以提高动物存活率。
试验结束后，每组挑选3只健康、接近平均体重的肉鸡称重并颈静脉放血处死，解剖取空肠组织采用ELISA分析免疫水平变化，结果见图2和图3。结果显示，与对照组相比，试验1组的免疫球蛋白IgA显著提升，炎症因子IL-1β水平显著降低；与试验2相比，试验3组的免疫球蛋白IgA显著提升，炎症因子IL-1β水平显著降低；表现出凝结芽孢杆菌BC66有提升免疫和降低炎症损伤的作用。
实施例7凝结芽孢杆菌BC66在水产养殖中的应用
试验用初始体重为10g左右黄颡鱼幼鱼300尾，采用循环流水控温系统进行养殖。本实验随机分为2组，每组3个平行，每个平行50尾鱼。其中，对照组：饲喂基础日粮；试验组：饲料是基础日粮基础上含有108CFU/g的凝结芽孢杆菌BC66菌粉制剂。饲养管理：试验前，先用基础饲料驯养10d后进行正式试验。试验期间，每天按照体重的3％-5％投喂，每天早晚各投喂1餐，养殖14d后进行称重，计算其增重率、特定生长率、饵料系数和存活率。
表3凝结芽孢杆菌BC66对黄颡鱼幼鱼生长性能的影响
注：同一列数据中肩标不同字母表示差异显著(P<0.05)，肩标差异字母或者无肩标表示差异不显著(P＞0.05)。
与对照组相比，添加有凝结芽孢杆菌BC 66的饲料饲喂后能够显著提高鱼体的增重率、特定生长率，并能够显著降低饵料系数；存活率均较高，基本上无死亡现象，无显著差异。养殖结果表明，益生菌凝结芽孢杆菌BC66能够显著提高黄颡鱼幼鱼的生长发育，调节鱼类肠道健康，提高饲料消化率。
Claims (10)
1.一种凝结芽孢杆菌BC66，其微生物保藏号为CGMCC21879；分类命名为：Bacilluscoagulans；保藏单位：中国微生物菌种保藏管理委员会普通微生物中心。
2.根据权利要求1所述的所述凝结芽孢杆菌BC66，其特征在于，其16SrRNA基因序列如SEQ ID No 1所示。
3.权利要求1所述凝结芽孢杆菌BC66的菌粉的制备方法，其特征在于：
凝结芽孢杆菌菌种从甘油管出发，接入试管斜面活化，经一级种子摇瓶培养后，按照5％接种量转入种子罐扩培，得到二级种子液；将二级种子液转入100L发酵罐发酵，得到的发酵液经12000rmp离心后收集菌泥，通过喷雾干燥工艺得到菌粉。
4.根据权利要求3所述的制备方法，其特征在于，所用的种子培养基配方以重量百分比计，葡萄糖2.0％、酵母浸粉1.0％、蛋白胨1.0％、MgSO4·7H2O 0.03％、MnSO4 0.01％、NaCl0.05％、K2HPO4 0.1％、FeSO4·7H2O 0.001％，CaCO3 0.05％，余量为水，调节初始pH值为7.0±0.1，115℃灭菌30min备用。
5.根据权利要求3或4所述的制备方法，其特征在于，所用的发酵培养基配方以重量百分比计，葡萄糖3.0％、酵母浸粉3.5％、蛋白胨4.0％、MgSO4·7H2O 0.06％、MnSO4 0.01％、NaCl 0.1％、K2HPO4 0.2％、FeSO4·7H2O 0.002％，CaCO30.1％，余量为水，调节初始pH值为7.0±0.1，115℃灭菌30min备用。
6.权利要求1所述的凝结芽孢杆菌BC66或其菌粉在畜牧养殖中作为饲料或药物的添加剂的应用。
7.根据权利要求6所述的应用，其特征在于，所述的饲料或药物包括用于猪、鸡、鸭、牛、羊或兔的饲料或药物。
8.根据权利要求6所述的应用，其特征在于，所述的饲料包括用于水产养殖的饲料或药物。
9.权利要求1所述的凝结芽孢杆菌BC66或其菌粉在制备畜牧养殖饲料或药物的添加剂中的应用。
10.权利要求1所述的凝结芽孢杆菌BC66或其菌粉在制备畜牧水产养殖饲料或药物中的应用。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110906213.0A CN113717881A (zh) | 2021-08-09 | 2021-08-09 | 一种凝结芽孢杆菌bc66及其应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110906213.0A CN113717881A (zh) | 2021-08-09 | 2021-08-09 | 一种凝结芽孢杆菌bc66及其应用 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113717881A true CN113717881A (zh) | 2021-11-30 |
Family
ID=78675160
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110906213.0A Pending CN113717881A (zh) | 2021-08-09 | 2021-08-09 | 一种凝结芽孢杆菌bc66及其应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113717881A (zh) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114437975A (zh) * | 2022-01-27 | 2022-05-06 | 黄河三角洲京博化工研究院有限公司 | 一株产乳酸凝结芽孢杆菌及其应用 |
| CN114686415A (zh) * | 2021-12-23 | 2022-07-01 | 微康益生菌(苏州)股份有限公司 | 凝结芽孢杆菌bc99在制备富硒药物或食品中的应用 |
| CN115094012A (zh) * | 2022-08-18 | 2022-09-23 | 北京农学院 | 凝结芽孢杆菌bc-hyc株菌剂的制备方法及应用 |
| CN115354002A (zh) * | 2022-09-20 | 2022-11-18 | 微康益生菌(苏州)股份有限公司 | 一种凝结芽孢杆菌bc07、其用途及其抑菌剂、药物、食品和抑菌方法 |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112852679A (zh) * | 2021-03-17 | 2021-05-28 | 武汉微康益生菌研究院有限公司 | 一株益生凝结芽孢杆菌及其应用 |
-
2021
- 2021-08-09 CN CN202110906213.0A patent/CN113717881A/zh active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112852679A (zh) * | 2021-03-17 | 2021-05-28 | 武汉微康益生菌研究院有限公司 | 一株益生凝结芽孢杆菌及其应用 |
Non-Patent Citations (1)
| Title |
|---|
| 陈明等: "凝结芽孢杆菌的生物学功能及其在动物生产中的应用", 《畜牧与兽医》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114686415A (zh) * | 2021-12-23 | 2022-07-01 | 微康益生菌(苏州)股份有限公司 | 凝结芽孢杆菌bc99在制备富硒药物或食品中的应用 |
| CN114686415B (zh) * | 2021-12-23 | 2023-06-16 | 微康益生菌(苏州)股份有限公司 | 凝结芽孢杆菌bc99在制备富硒药物或食品中的应用 |
| CN114437975A (zh) * | 2022-01-27 | 2022-05-06 | 黄河三角洲京博化工研究院有限公司 | 一株产乳酸凝结芽孢杆菌及其应用 |
| CN114437975B (zh) * | 2022-01-27 | 2023-09-29 | 黄河三角洲京博化工研究院有限公司 | 一株产乳酸凝结芽孢杆菌及其应用 |
| CN115094012A (zh) * | 2022-08-18 | 2022-09-23 | 北京农学院 | 凝结芽孢杆菌bc-hyc株菌剂的制备方法及应用 |
| CN115354002A (zh) * | 2022-09-20 | 2022-11-18 | 微康益生菌(苏州)股份有限公司 | 一种凝结芽孢杆菌bc07、其用途及其抑菌剂、药物、食品和抑菌方法 |
| CN115354002B (zh) * | 2022-09-20 | 2023-12-19 | 微康益生菌(苏州)股份有限公司 | 一种凝结芽孢杆菌bc07、其用途及其抑菌剂、药物、食品和抑菌方法 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102373172B (zh) | 一株屎肠球菌及其应用 | |
| CN103614327B (zh) | 一种枯草芽孢杆菌及其用途 | |
| CN113717881A (zh) | 一种凝结芽孢杆菌bc66及其应用 | |
| CN110982760B (zh) | 一株贝莱斯芽孢杆菌及其在防治猪腹泻中的应用 | |
| CN103981118B (zh) | 一种枯草芽孢杆菌饲料添加剂及其制备方法和应用 | |
| CN112011481B (zh) | 一株防治畜禽细菌性腹泻的罗伊氏乳杆菌及其应用 | |
| CN102559539B (zh) | 嗜酸乳杆菌及其应用与饲料添加剂和预混料 | |
| US20090257995A1 (en) | Harmful bacterium control agent containing bacillus thuringiensis | |
| CN109161509B (zh) | 一株能防治牛羊腹泻病的菌株 | |
| CN110878267A (zh) | 一株唾液乳杆菌及其用途 | |
| CN114085789B (zh) | 戊糖片球菌ma.wtpqj01及其应用 | |
| CN109136126B (zh) | 一株用于预防断奶仔猪腹泻的戊糖片球菌 | |
| CN104171429A (zh) | 一种植物乳杆菌lp28的用途 | |
| WO2023050716A1 (zh) | 一种融合魏斯氏菌、培养方法及其应用 | |
| KR20200034169A (ko) | 항균 활성, 항노화 활성 및 프로바이오틱스 특성을 가지는 락토바실러스 플란타룸 jdfm lp11 균주 및 이의 용도 | |
| CN115094012A (zh) | 凝结芽孢杆菌bc-hyc株菌剂的制备方法及应用 | |
| KR20090066410A (ko) | 바실러스 서브틸리스 ｂｃ1212 균주를 포함하는 사료첨가용 조성물 | |
| CN115505546A (zh) | 一株抑制嗜水气单胞菌的耐盐芽孢杆菌及其应用 | |
| CN113025534B (zh) | 一株具有较强抑菌作用的新种芽孢杆菌x901株及其应用 | |
| CN113502243B (zh) | 一株高产乳酸的植物乳杆菌gbw-lp001及其替抗菌剂和应用 | |
| KR20150117184A (ko) | 신규한 유산균, 진균 및 세균을 포함하는 사료첨가용 조성물 | |
| KR101854705B1 (ko) | 알파글루코시다아제 저해제를 다량 생산하는 바실러스 리케니포르미스 ny1505 균주 | |
| CN115572689A (zh) | 一株戊糖片球菌m6及其应用 | |
| GB2484126A (en) | Foodstuff fermented with lactic acid producing bacteria | |
| CN114657082B (zh) | 一种乳酸乳球菌及其应用 |
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 | ||
| TA01 | Transfer of patent application right | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20240424 Address after: 462000 Southeast corner of the intersection of Longjiang Road and Wutaishan Road, Yancheng District, Luohe City, Henan Province Applicant after: Henan Anmaikang Biotechnology Co.,Ltd. Country or region after: China Address before: No. 1033, Longqiao Road, Wujiang Economic and Technological Development Zone, Wujiang District, Suzhou, Jiangsu 215222 Applicant before: Weikang probiotics (Suzhou) Co.,Ltd. Country or region before: China |